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2015 Vol. 31, No. 5

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Expert Forum
Repair of skin and soft tissue defects around the knee joints
Tan Qian, Xu Peng
2015, 31(5): 321-324. doi: 10.3760/cma.j.issn.1009-2587.2015.05.001
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Skin and soft tissue defects around the knee joints are often accompanied by popliteal artery injury, patellar ligament injury, patellar fracture, and other deep tissue damage or exposure, making them challenging to repair. The principle is to repair the wound, reconstruct anatomical structure of the knee joint, and recover the knee joint function. At present the reconstruction with skin flap or myocutaneous flap is our priority. Local flap or myocutaneous flap can be used for repairing minor defects around the knee joints. Repairing with perforator flap, fascia flap, and free flap are main alternatives for covering larger and complex defects around the knee joints. During the treatment, a joint effort is mandatory, not only to repair the wound, but also to reconstruct vasculature, fix fracture, repair ligament, and finally recover the knee joint function. Therefore, the importance of multidisciplinary cooperation must be emphasized. Moreover, along with the development of new technologies, new methods, and new materials, perforator flap plays an important role in repairing skin and soft tissue defects around the knee joints.
2015, 31(5): 324-324. doi: 10.3760/cma.j.issn.1009-2587.2015.05.104
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2015, 31(5): 324-324. doi: 10.3760/cma.j.issn.1009-2587.2015.05.103
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Repair of skin and soft tissue defects around the knee joints combined with patellar ligament defects using free anterolateral thigh flaps with iliotibial tracts
Han Fu, Hu Dahai, Liu Yang, Yu Hongliang, Ma Shaojun, Wei Guoxing, Zheng Zhao
2015, 31(5): 327-330. doi: 10.3760/cma.j.issn.1009-2587.2015.05.003
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Objective To observe clinical efficacy of using free anterolateral thigh flaps with iliotibial tracts in repairing skin and soft tissue defects around the knee joints with patellar ligament defects. Methods Twelve patients with skin and soft tissue defects around the knee joints and patellar ligament defects were hospitalized from June 2010 to June 2014. The defects of skin and soft tissue ranged from 7 cm×6 cm to 16 cm×12 cm in area, and patellar ligament ranged from 5 to 12 cm in length and 2.5 to 4.0 cm in width. Free anterolateral thigh flaps with iliotibial tracts were used to repair these defects. During reconstruction of patellar ligament, both ends of iliotibial tract were successively folded to form tendon-like three-layer structure at first, and then the newly formed structure was wrapped around the broken ends of patellar ligament and fixed with suture. The flap size ranged from 9 cm×8 cm to 18 cm×14 cm. The iliotibial tract ranged from 7 to 14 cm in length and 8 to 12 cm in width. The donor sites were closed by grafting with autologous split-thickness skin harvested from thigh or trunk, and parked with gauze. Immediately after operation, the knee joints were fixed in extension with orthosis for 6 weeks. Weight bearing training of affected limbs being kept in extension position was started from 2 weeks after operation, and flexion and extension exercise of affected knee joints was begun from 6 weeks after operation. Before operation and 12 months after operation, the degree of pain around the knee joints and knee joint function were evaluated with the international knee documentation committee knee uation form, and the ranges of flexion and extension of knee joints were also evaluated. The integrity of reconstructed patellar ligament was assessed by color Doppler ultrasound from 6 to 12 months after operation. The occurrence of surgery-related complications was observed in all patients within 12 months after operation. Results (1) After operation, all flaps survived well, and all wounds healed well. (2) The average score of pain around the knee joint was increased from 31 points before operation to 77 points in 12 months after operation. The average score of knee joint function was increased from 14 points before operation to 65 points in 12 months after operation. Before operation, the average ranges of flexion and extension of knee joint were respectively 89° and 65°, and they were respectively increased to 130° and decreased to 15° in 12 months after operation. From 6 to 12 months after operation, color Doppler ultrasound showed that the condition of reconstructive patellar ligaments in all patients was good without the need for further surgical intervention; the superficial sensation of the flaps was recovered in different degrees. No surgery-related complication was observed in all patients within 12 months after operation. Conclusions Free grafting of anterolateral thigh flap with iliotibial tract is an effective and reliable method for repairing skin and soft tissue defects around the knee joints combined with patellar ligament defects, and the surgical procedure can recover function and appearance of knee joint satisfactorily.
Tissue flap repair strategy for severe defects of skin and soft tissue around the knee joints
Shen Yuming, Ma Chunxu, Hu Xiaohua, Wang Cheng, Zhang Cong
2015, 31(5): 331-336. doi: 10.3760/cma.j.issn.1009-2587.2015.05.004
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Objective To explore selection and method of tissue flaps for the repair of severe defects of skin and soft tissue around the knee joints. Methods Fifty-four patients with wounds around the knee joints, all accompanied by exposure or necrosis of tendon or bone and exposure of prosthesis, were hospitalized in our burn center from June 2008 to December 2014. Five of them were with knee joint injury. After thorough debridement or tumor resection, the wound area ranged from 5 cm×5 cm to 46 cm×22 cm. Three patients were repaired with free latissimus dorsi myocutaneous flaps, 7 were repaired with modified sartorius myocutaneous flaps, 8 were repaired with gastrocnemius myocutaneous flaps, one was repaired with gastrocnemius muscle flap, two were repaired with posterior leg flaps combined with gastrocnemius muscle flaps, one was repaired with femoral biceps muscle flap combined with gastrocnemius muscle flap, 13 were repaired with reverse anterolateral thigh island flaps, two were repaired with reverse anterolateral thigh island flap combined with gastrocnemius myocutaneous flaps, two were repaired with superior lateral genicular flaps, 4 were repaired with reverse posterior thigh island flaps, 11 were repaired with saphenous artery flaps. Patellar ligament was reconstructed in 4 patients. The tissue flap size ranged from 5 cm×5 cm to 38 cm×19 cm. Some donor sites were sutured directly, and the others were closed by split-thickness skin grafting obtained from ipsilateral or contralateral legs. Results Among 59 tissue flaps of 54 patients, 55 tissue flaps of 50 patients survived, while necrosis of the distal part was observed in 4 tissue flaps, including one saphenous artery flap, two reverse anterolateral thigh island flaps, and one free latissimus dorsi myocutaneous flap. Among them, 3 flaps with necrosis at the distal part healed after debridement followed by skin grafting, one myocutaneous flap healed by transplanting gastrocnemius myocutaneous flap. During the follow-up period of 6 to 36 months, the tissue flaps were in good appearance and texture, and knee joint function was good in most cases. In 4 patients the knee joint function was satisfactory after patellar ligament reconstruction, while stiffness was observed in 4 out of 5 patients with knee joint injury. Conclusions Free latissimus dorsi myocutaneous flaps are preferred to repair extensive defects around the knee joints. Reverse anterolateral thigh island flaps followed by saphenous artery flaps are preferred to repair wounds around the anterior knee. Wounds of the lateral knee are mainly repaired with reverse anterolateral thigh island flaps, and for small wounds the use of the superior lateral genicular flaps may be considered. Wounds of the medial knee can be repaired with modified sartorius myocutaneous flaps or saphenous artery flaps. Wounds of the posterior knee can be repaired with reverse posterior thigh island flaps or superior lateral genicular flaps. Wounds with severe infection or large space can be repaired with gastrocnemius myocutaneous flaps or muscle flaps or modified sartorius myocutaneous flaps. Anterolateral thigh flaps and gastrocnemius myocutaneous flaps are preferred in cases with indication of patellar ligament reconstruction.
2015, 31(5): 336-336. doi: 10.3760/cma.j.issn.1009-2587.2015.05.101
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2015, 31(5): 336-336. doi: 10.3760/cma.j.issn.1009-2587.2015.05.102
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Repair of large and deep skin and soft tissue defects around the knee joints with free latissimus dorsi musculocutaneous flaps
Zhang Minghua, Cui Xu, Zeng Jizhang, Liu Xiong, Huang Mitao, Zhang Pihong, Huang Xiaoyuan
2015, 31(5): 337-339. doi: 10.3760/cma.j.issn.1009-2587.2015.05.005
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Objective To investigate the clinical efficacy of free latissimus dorsi musculocutaneous flaps in repairing large and deep skin and soft tissue defects around the knee joints. Methods Twenty-five patients with large and deep skin and soft tissue defects around the knee joints were hospitalized from March 2005 to March 2014. The area of defects around the knee joints ranged from 10 cm×8 cm to 43 cm×23 cm. The free latissimus dorsi musculocutaneous flaps were used to repair the defects, with the area ranging from 12 cm×10 cm to 45 cm×25 cm. The thoracodorsal artery and its concomitant vein of the musculocutaneous flap were anastomosed to the descending branch of the lateral circumflex femoral artery and its concomitant vein respectively to reconstruct blood supply. Split-thickness skin grafts around the flap donor sites were harvested to cover the muscle surface of the musculocutaneous flaps. The flap donor sites were closed directly with suture, and the skin donor sites were healed by dressing change. Results All the 25 flaps survived without vascular crisis. The flaps were in satisfactory appearance. The flap donor sites were healed with linear scar. All the patients were followed up for 3 to 6 months. At last, they were able to stand up and walk. Conclusions The free latissimus dorsi musculocutaneous flap transplantation is an effective treatment for the repair of large and deep soft tissue defects around the knee joints, and the descending branch of lateral circumflex femoral artery and its concomitant vein are the appropriate recipient vessels.
2015, 31(5): 340-341. doi: 10.3760/cma.j.issn.1009-2587.2015.05.006
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2015, 31(5): 341-344. doi: 10.3760/cma.j.issn.1009-2587.2015.05.007
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2015, 31(5): 378-380. doi: 10.3760/cma.j.issn.1009-2587.2015.05.014
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2015, 31(5): 381-383. doi: 10.3760/cma.j.issn.1009-2587.2015.05.015
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2015, 31(5): 384-385. doi: 10.3760/cma.j.issn.1009-2587.2015.05.016
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2015, 31(5): 386-387. doi: 10.3760/cma.j.issn.1009-2587.2015.05.017
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2015, 31(5): 388-388. doi: 10.3760/cma.j.issn.1009-2587.2015.05.018
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2015, 31(5): 392-393. doi: 10.3760/cma.j.issn.1009-2587.2015.05.020
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2015, 31(5): 393-395. doi: 10.3760/cma.j.issn.1009-2587.2015.05.021
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2015, 31(5): 396-397. doi: 10.3760/cma.j.issn.1009-2587.2015.05.022
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2015, 31(5): 398-399. doi: 10.3760/cma.j.issn.1009-2587.2015.05.023
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2015, 31(5): 399-400. doi: 10.3760/cma.j.issn.1009-2587.2015.05.024
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Expert Comment
To further improve the techniques for repair and reconstruction of skin and soft tissue defects around the knee joints
Huang Xiaoyuan
2015, 31(5): 325-326. doi: 10.3760/cma.j.issn.1009-2587.2015.05.002
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This article briefly summarizes the techniques for repair of skin and soft tissue defects around the knee joints as reported in 5 papers in this issue, including how to choose the skin flap, muscle flap, myocutaneous flap, and vascular anastomosis in recipient site. It is found that the anterolateral femoral flap, latissimus dorsi myocutaneous flap, and gastrocnemius flap are widely used in clinic with high survival rates, and they can be used for the repair of large soft tissue defects as well as the reconstruction of the knee joint function.
Original Article
Efficacy observation on repair of finger pulp defects and sensory reconstruction of children with the perforator propeller flaps based on the end dorsal branch of digital proper artery in the same finger
Feng Shiming, Wang Aiguo, Zhang Zaiyi, Tao Youlun, Zhou Mingming, Hao Yunjia, Sun Qingqing
2015, 31(5): 345-348. doi: 10.3760/cma.j.issn.1009-2587.2015.05.008
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Objective To investigate the clinical outcomes of the use of the perforator propeller flaps based on the end dorsal branch of digital proper artery in the same finger in repair of finger pulp defects and sensory reconstruction in children. Methods Twenty-three children (31 fingers) with index, middle, ring or little finger pulp defects were hospitalized from September 2012 to December 2013. The area of finger pulp defects ranged from 1.2 cm×1.0 cm to 2.0 cm×1.5 cm. The perforator propeller flaps based on the end dorsal branch of digital proper artery in the same finger were used to repair the defects, with the flap size ranging from 1.3 cm×1.2 cm to 2.2 cm×1.6 cm. The dorsal branch of the digital proper nerve of the flap was conducted end-to-end anastomosis with the broken end of the nerve of the wound to reconstruct sensation. The donor sites were covered with autologous full-thickness skin obtained from inner aspect of the thigh. Results Primary healing of the wounds and donor sites were achieved in all 23 children. All the flaps and skin grafts of donor sites survived. All the patients were followed up for 6 to 20 months, with mean time of 14 months. At the last follow-up, the flaps and donor sites were in good appearance, the finger pulps were mellow and plump, with no obvious pigmentation or cicatricial contracture. The sensation of finger pulps reached S3+, and the distance of two-point discrimination ranged from 4.5 to 6.0 mm, with mean distance of 5.1 mm. Twenty-one parents of the patients were strongly satisfied with the appearance of the repaired fingers, and the other 2 parents also expressed satisfaction. Conclusions Transplantation of the perforator propeller flap based on the end dorsal branch of digital proper artery in the same finger is a safe and reliable method for the repair of index, middle, ring, and little finger pulp defects and sensory reconstruction of children. The flaps are with good blood supply, appearance and sensory function after operation.
Fibrosis after damage to fat dome structure of skin of pig
Yu Xiaoping, Kang Yutian, Zuo Yanhai, Liu Chuanbo, Ye Junna, Yuan Bo, Ji Xiaoyun, Song Fei, Jiang Yuzhi, Xiao Yurui, Jin Shuwen, Lu Shuliang
2015, 31(5): 349-353. doi: 10.3760/cma.j.issn.1009-2587.2015.05.009
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Objective To observe the fibrosis of skin after damage to the fat dome structure in skin of pig. Methods Totally 4 pieces of skin grafts of intermediate thickness in the size of 5 cm×5 cm were obtained from both sides beside the spine of back in each of the 4 female red Duroc pigs with pedicle on one side with Humby knife performed by burn specialists, who were rich in clinical experience. These skin grafts were assigned as thin dermis group (TD). Pedicled tissue grafts in the size of 5 cm×5 cm with the thickness of 1.5 mm were obtained within the wounds resulted from former incision with the same method mentioned above, and these tissue grafts were set as fat dome group (FD). The above-mentioned two groups of skin grafts were sutured back in situ immediately after completion of the former procedures. On post surgery day (PSD) 7, 14, and 21, 5 wounds were respectively selected according to the random number table for gross observation of the surgical areas. Tissue samples were obtained from corresponding surgical area deep to the deep fascia after gross observation at above-mentioned time points. Some of the tissue samples were used for observation of distribution of collagen fibers in the regions of operation of both groups of skin grafts with HE staining, and the breadth of fibrosis was measured; some of the tissue samples were used for observation of distribution of type Ⅰ or Ⅲ collagen fibers in the regions of incision of both two groups of skin grafts with Sirius red staining. Data were processed with two independent sample t test. Results A little scab on the edge of wounds was observed on PSD 7; all the wounds were healed on PSD 14; a few hairs were observed growing in the surgical area on PSD 21. HE staining showed that traces of incision were observed in the superficial layer of dermis and at the junction between dermis and fat dome at each time point; profuse hyperplasia of collagen fibers with parallel and orderly arrangement were observed in the region of incision of skin grafts in groups TD and FD at each time point. The breadth of fibrosis of the region of incision of skin grafts was respectively (251±31), (240±37), and (342±69) μm in group TD, (239±36), (286±61), and (332±28) μm in group FD on PSD 7, 14, 21, without significantly statistical difference (with t values respectively 0.750, -1.971, and 0.375, P values above 0.05). Sirius red staining showed that large amount of type Ⅲ collagen fibers and small amount of type Ⅰ collagen fibers arranging parallelly were present in the region of incision of skin grafts in groups TD and FD at each time point. Conclusions Under the circumstances of relatively intact restoration of dermal tissue, no excessive fibrosis was observed after simple incisional injury of fat dome in skin of pig.
Study of exogenous carbon monoxide-releasing molecules 2 on endotoxin/lipopolysaccharide-induced abnormal activation of platelets of healthy human donors
Liu Dadong, Zhuang Mingfeng, Zhang Jingli, Chen Jingjia, Sun Bingwei
2015, 31(5): 354-360. doi: 10.3760/cma.j.issn.1009-2587.2015.05.010
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Objective To explore the effects of exogenous carbon monoxide-releasing molecules 2 (CORM-2) on LPS-induced abnormal activation of platelets in peripheral blood of healthy human donors and its possible molecular mechanism. Methods Venous blood samples were collected from a healthy volunteer, and platelet-rich plasma (PRP) from the blood were isolated by differential centrifugation. The PRP was subpackaged into siliconized test tubes and then divided into control group, LPS group, inactive CORM-2 (iCORM-2) group, 10 μmol/L CORM-2 group, and 50 μmol/L CORM-2 group according to the random number table, with 3 tubes in each group. The PRP in control group did not receive any treatment. The PRP in LPS group received LPS (20 mL, 10 μg/mL) stimulation, and the PRP in iCORM-2 group, 10 μmol/L CORM-2 group, and 50 μmol/L CORM-2 group underwent the same LPS stimulation and treatment of 50 μmol/L iCORM-2, 10 μmol/L CORM-2, and 50 μmol/L CORM-2, respectively, with the dosage of 20 mL. After being cultured for 30 min, the platelet adhesion rate was determined by glass bottle method, the number of platelet spreading on fibrinogen was determined with immunofluorescent method, and the platelet aggregation rate was measured by turbidimetric method. The platelet poor plasma (PPP) was prepared from PRP, the levels of ATP in PPP and platelets were determined by chemical fluorescein method. The expressions of platelet glycoprotein Ⅰ bα (GPⅠbα) and GPⅥ were analyzed by flow cytometer. The expressions of glycogen synthase kinase 3β (GSK-3β) and phosphorylated GSK-3β were determined by Western blotting and immunoprecipitation, respectively. Measurement of the above indices was repeated for 3 times. Data were processed with one-way analysis of variance and SNK test. Results Compared with those in control group, the platelet adhesion rates, numbers of platelets spreading on fibrinogen, platelet aggregation rates, expressions of GPⅠbα and GPⅥ in PRP, levels of ATP in PPP in LPS and iCORM-2 groups were significantly increased, while levels of ATP in platelets were significantly decreased (with P values below 0.05). Compared with those in LPS group, the former 7 indices in iCORM-2 group showed no significant differences (with P values above 0.05), while the levels of ATP in platelets in the 10 μmol/L CORM-2 and 50 μmol/L CORM-2 groups were significantly increased, and the other 6 indices in 10 μmol/L CORM-2 and 50 μmol/L CORM-2 groups were significantly decreased (with P values below 0.05). The expression levels of GSK-3β of the platelets in PRP in control, LPS, iCORM-2, 10 μmol/L CORM-2, and 50 μmol/L CORM-2 groups were 0.550±0.060, 1.437±0.214, 1.210±0.108, 0.720±0.010, and 0.670±0.010, respectively, and the expression levels of the phosphorylated GSK-3β of the platelets in PRP in the above 5 groups were 0.950±0.070, 1.607±0.121, 1.420±0.040, 1.167±0.015, and 0.513±0.122, respectively. Compared with those in control group, both the expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in LPS and iCORM-2 groups were significantly increased (with P values below 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP between LPS group and iCORM-2 group were similar (with P values above 0.05). The expression levels of GSK-3β and phosphorylated GSK-3β of the platelets in PRP in 10 μmol/L CORM-2 and 50 μmol/L CORM-2 groups were significantly decreased compared with those in LPS group (with P values below 0.05). Conclusions LPS stimulation can abnormally activate the platelets in peripheral blood of healthy human, but the abnormal activation can be inhibited by CORM-2 intervention, and the mechanism of the latter may involve the phosphorylation of GSK-3β mediated by GP.
Effects of zinc deficiency on the relevant immune function in rats with sepsis induced by endotoxin/lipopolysaccharide
Li Feng, Cong Tao, Li Zhen, Zhao Lin
2015, 31(5): 361-366. doi: 10.3760/cma.j.issn.1009-2587.2015.05.011
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Objective To investigate the effects of zinc deficiency on the relevant immune function in rats with LPS-induced sepsis. Methods Sixty rats were divided into low zinc group (LZ), normal zinc pair-fed group (NP), and normal zinc control group (NC) according to the random number table, with 20 rats in each group. The rats in group LZ were fed with low zinc diet, and the rats in group NP were fed with normal zinc diet, with the same intake as that of group LZ by manual control, and the rats in group NC were fed with normal zinc diet freely. After being fed for 7 d, the rats all fasted and were further divide into the below subgroups named LZ-LPS, LZ-normal saline (NS), NP-LPS, NP-NS, NC-LPS, and NC-NS according to the random number table, with 10 rats in each subgroup. Rats in the LPS subgroups were intraperitoneally injected with 1 mg/mL LPS solution with the dosage of 5 mg/kg, rats in the corresponding NS subgroups were intraperitoneally injected with equivalent NS. The rats were sacrificed at post injection hour 6 to collect blood, spleen, and thymus. The serum level of zinc was detected by inductively coupled plasma mass spectrometry, and the serum alkaline phosphatase (ALP) activity was detected by automatic blood biochemical analyzer. The body weight and weight of spleen and thymus of rats were weighed, and the indices of spleen and thymus were calculated. Six routine blood indices were examined by automatic blood cell analyzer. The serum levels of interferon gamma (IFN-γ), TNF-α, IL-4, and IL-10 were determined with ELISA, and the ratio of IFN-γ to IL-4 was calculated. Data were processed with one-way analysis of variance and SNK test. Results (1) Serum levels of zinc and ALP activity in the LPS subgroups were significantly lower than those in the corresponding NS subgroups (with P values below 0.05). The two former indices in subgroups NP-NS and NC-NS were significantly higher than those in subgroup LZ-NS (with P values below 0.05). The two former indices in subgroups NP-LPS and NC-LPS were significantly higher than those in subgroup LZ-LPS (with P values below 0.05). (2) Body weight, spleen and thymus weight, indices of spleen and thymus in the LPS subgroups were similar with those in the corresponding NS subgroups (with P values above 0.05). The 4 former indices, except for body weight, in subgroups NP-NS and NC-NS were significantly higher than those in subgroup LZ-NS (with P values below 0.05). The 4 former indices, except for body weight, in subgroups NP-LPS and NC-LPS were significantly higher than those in subgroup LZ-LPS (with P values below 0.05). (3) Levels of leucocyte count in subgroups LZ-LPS and NP-LPS were significantly higher than those in the corresponding NS subgroups (with P values below 0.05). Level of leucocyte count in subgroup NC-NS was significantly higher than that in subgroup LZ-NS (P<0.05). Level of leucocyte count in subgroup NC-LPS was significantly lower than that in subgroup LZ-LPS (P<0.05). Levels of neutrophilic granulocyte count (NGC) and NG in the LPS subgroups were significantly higher than those in the corresponding NS subgroups (with P values below 0.05). The two former indices in subgroup NC-LPS were significantly lower than those in subgroup LZ-LPS (with P values below 0.05). Level of NG in subgroup NC-NS was significantly lower than that in subgroup LZ-NS (P<0.05). Levels of lymphocyte count and lymphocyte in subgroups LZ-NS, LZ-LPS, NP-NS, NP-LPS, NC-NS, and NC-LPS were respectively (1.8±0.4)×109/L, (1.0±0.3)×109/L, (2.6±0.7)×109/L, (1.4±0.4)×109/L, (3.3±0.6)×109/L, (1.5±0.5)×109/L, and 0.39±0.10, 0.11±0.03, 0.47±0.12, 0.14±0.04, 0.50±0.09, 0.24±0.07. The two former indices in the LPS subgroups were significantly lower than those in the corresponding NS subgroups (with P values below 0.05). The two former indices in subgroup NC-NS were significantly higher than those in subgroup LZ-NS (with P values below 0.05). The two former indices in subgroups NP-LPS and NC-LPS were significantly higher than those in subgroup LZ-LPS (with P values below 0.05). Level of lymphocyte count in subgroup NP-NS was significantly higher than that in subgroup LZ-NS (P<0.05). Levels of platelet count (PC) in subgroups NP-LPS and NC-LPS were significantly lower than those in the corresponding NS subgroups (with P values below 0.05). Levels of PC in subgroups NP-NS and NC-NS were significantly higher than those in subgroup LZ-NS (with P values below 0.05). Level of PC in subgroup NC-LPS was significantly higher than that in subgroup LZ-LPS (P<0.05). (4) Serum levels of TNF-α, IL-4, and IL-10 in each subgroup showed no significant differences (with P values above 0.05). Serum levels of IFN-γ and ratios of IFN-γ to IL-4 in subgroups LZ-NS, LZ-LPS, NP-NS, NP-LPS, NC-NS, and NC-LPS were respectively (75±21), (233±40), (80±14), (345±74), (66±7), (821±189) pg/mL, and 3.1±1.0, 6.6±1.7, 3.9±1.7, 20.2±8.3, 3.4±1.5, 45.7±7.6. The two former indices in the LPS subgroups were significantly higher than those in the corresponding NS subgroups (with P values below 0.05). The two former indices in subgroups NP-NS and NC-NS were similar with those in subgroup LZ-NS (with P values above 0.05). The two former indices in subgroups NP-LPS and NC-LPS were significantly higher than those in subgroup LZ-LPS (with P values below 0.05). Conclusions Zinc deficiency can induce the atrophy of spleen and thymus, and reduction of peripheral blood lymphocyte. In sepsis, zinc deficiency can further decrease the production of IFN-γ, thus making the cytokines of Th1/Th2 shift to Th2 and the immune imbalance worse.
Expression of microRNA-126 in myocardial tissue of rats in the early stage of severe burn injury and its relation with myocardial damage
Xie Qionghui, Ye Ziqing, Chen Lan, Zhao Chaoli, Ruan Qiongfang, Xie Weiguo
2015, 31(5): 367-371. doi: 10.3760/cma.j.issn.1009-2587.2015.05.012
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Objective To observe the changes in the expressions of microRNA-126 in myocardial tissue and cardiac troponin I (cTnI) in serum of rats in the early stage of severe burn injury with analysis of their relationship, and to validate the relationship between microRNA-126 and myocardial damage in cellular level. Methods (1) Forty-eight SD rats were divided into sham injury group (n=8, without fluid therapy after sham injury) and burn injury group (n=40, inflicted with 30% TBSA full-thickness scald on the back, hereinafter referred to as burn, and received intraperitoneally injection of lactic acid Ringer's solution) according to the random number table. Blood was collected from abdominal aorta of rats in sham injury group at post injury hour (PIH) 1, and then these 8 rats were sacrificed for obtaining left ventricular tissue. Blood was respectively collected from abdominal aorta of 8 rats in burn injury group at PIH 3, 6, 12, 24, and 48, and then they were sacrificed and the left ventricular tissue was obtained at each time point. The expression of microRNA-126 in myocardial tissue was assessed by real-time fluorescent quantitative RT-PCR. Serum level of cTnI was assessed by ELISA. (2) Rat myocardial cell line H9C2 was divided into normal control group (NC, routinely cultured), stimulation group (S), negative transfection+ stimulation group (NT+ S), and transfection+ stimulation group (T+ S) according to the random number table. Cells in S group were treated with hypoxia for 24 h after being cultured with DMEM containing 10% burn serum obtained from rats in burn injury group at PIH 6 in experiment (1). Cells in NT+ S group and T+ S group were respectively transfected with the negative control of microRNA mimics and microRNA-126 mimics for 24 h, and then were given the same treatment as that of S group. The expression of microRNA-126 in myocardial cells was determined by real-time fluorescent quantitative RT-PCR (with the sample number of 3). Cell counting kit 8 was used to examine the vitality of myocardial cell (with the sample number of 4, denoted as absorbance value). Apoptotic rate of myocardial cells was determined by flow cytometer (with the sample number of 3). Data were processed with one-way analysis of variance and LSD-t test. The relationship between microRNA-126 expression in myocardial tissue and serum level of cTnI of rats was assessed by linear correlation analysis. Results (1) Compared with that of sham injury group at PIH 1, the expression levels of microRNA-126 in myocardial tissue of rats in burn injury group at PIH 3, 6, 12, 24, and 48 were significantly decreased (with t values from 5.68 to 9.79, P values below 0.01), reaching its nadir at PIH 24 (0.40±0.08). Compared with that of sham injury group at PIH 1, the serum levels of cTnI of rats in burn injury group at PIH 3, 6, 12, 24, and 48 were significantly increased (with t values from 6.68 to 12.79, P values below 0.01), peaking at PIH 12 [(1 035±177) pg/mL]. A significant negative correlation between the expression level of microRNA-126 in myocardial tissue and serum level of cTnI was observed in rats of burn injury group at each time point (r=-0.797, P<0.001). (2) Compared with those of NC group, the microRNA-126 expression levels in myocardial cells of S group and T+ S group were respectively decreased and increased (with t values respectively 4.57 and 5.73, P<0.05 or P<0.01), the cell vitality levels were obviously decreased (with t values respectively 14.88 and 6.48, P values below 0.01), and the apoptotic rates were significantly increased (with t values respectively 13.82 and 6.96, P values below 0.01). Compared with that in NT+ S group, the microRNA-126 expression level in myocardial cells of T+ S group was significantly increased (t=6.77, P<0.01), the cell vitality level was obviously increased (t=8.23, P<0.001), and the apoptotic rate was significantly decreased (t=6.14, P<0.001). Conclusions Expression level of microRNA-126 in myocardial tissue of rat was decreased in the early stage of severe burn injury. It may participate in regulating myocardial damage and play a protective role.
Effects of blocking two sites of transforming growth factor-β/Smads signaling on the formation of scar-related proteins in human skin fibroblasts
Wang Yang, Zhang Liangping, Lei Rui, Shen Yichen, Shen Hui, Wu Zhinan, Xu Jinghong
2015, 31(5): 372-377. doi: 10.3760/cma.j.issn.1009-2587.2015.05.013
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Abstract:
Objective To explore the effects of blocking two sites of TGF-β/Smads signaling on the formation of scar-related proteins in human skin fibroblasts. Methods Two lentivirus vectors encoding soluble TGF-β receptor Ⅱ (sTβRⅡ) and mutant Smad 4–Smad 4ΔM4 were respectively transfected into human skin fibroblast cell line human foreskin fibroblast 1 (HFF-1) cells with the optimum multiplicity of infection (MOI) of 50. The protein expressions of sTβRⅡ and Smad 4ΔM4 of the two types of transfected cells were determined by Western blotting so as to compare with those of the untransfected cells. The HFF-1 cells were divided into 6 groups as named below according to the random number table, with 6 dishes in each group, 1×104 cells per dish. Co-transfection group, transfected with the two previous lentivirus vectors, mixed with the ratio of 1∶1 and MOI of 50, and then stimulated with 5 ng/mL TGF-β1 for 72 h; sTβRⅡ group, transfected with lenti-sTβRⅡ with MOI of 50, with the other treatment as above; Smad 4ΔM4 group, transfected with lenti-Smad 4ΔM4 with MOI of 50, with the other treatment as above; negative virus group, transfected with empty lentivirus vector, with the other treatment as above; positive control group, stimulated with 5 ng/mL TGF-β1 for 72 h; and blank control group, conventionally cultured without any other treatment. After stimulation, Western blotting and real-time fluorescent quantitative RT-PCR were respectively used to determine the protein and mRNA expressions of fibronectin in cells of each group. ELISA and Sircol collagen assay were respectively used to determine the protein expressions of connective tissue growth factor (CTGF) and total collagen in the cell culture supernate of each group. Data were processed with one-way analysis of variance and SNK-q test. Results (1) HFF-1 cells transfected with lenti-sTβRⅡ and HFF-1 cells transfected with lenti-Smad 4ΔM4 respectively expressed higher levels of sTβRⅡ protein and Smad 4ΔM4 protein compared with those of untransfected cells, confirming that HFF-1 cells transfected with the two lentivirus vectors can efficiently express the target proteins. (2) There were statistically significant differences in the protein and mRNA expressions of fibronectin in cells of the 6 groups (with F values respectively 53.536 and 24.365, P values below 0.001). The protein and mRNA expressions of fibronectin in cells of positive control group (respectively 1.60±0.18 and 1.99±0.40) were similar with those of negative virus group (respectively 1.60±0.15 and 1.94±0.28, with q values respectively 0.091 and 0.419, P values above 0.05), and they were significantly higher than those of the rest 4 groups (with q values from 5.245 to 18.228, P values below 0.05). The protein and mRNA expressions of fibronectin in cells of co-transfection group (respectively 0.60±0.05 and 0.70±0.11) were significantly lower than those of sTβRⅡ group (respectively 0.89±0.13 and 1.24±0.17) and Smad 4ΔM4 group (respectively 0.91±0.14 and 1.28±0.19, with q values from 3.964 to 4.294, P values below 0.05). (3) There were statistically significant differences in the protein expressions of CTGF and total collagen in the cell culture supernate of the 6 groups (with F values respectively 107.680 and 38.347, P values below 0.001). The protein expressions of CTGF and total collagen in the cell culture supernate of positive control group were similar with those of negative virus group (with q values respectively 1.106 and 0.491, P values above 0.05), and they were significantly higher than those of the rest 4 groups (with q values from 6.414 to 26.420, P values below 0.05). The protein expressions of CTGF and total collagen in the cell culture supernate of co-transfection group were significantly lower than those of sTβRⅡ group and Smad 4ΔM4 group (with q values from 3.424 to 7.143, P values below 0.05). Conclusions In human skin fibroblasts, blockage of two sites of TGF-β/Smads signaling can reduce the expression of scar related proteins which are up-regulated by TGF-β1 to a greater extent than that of blocking one single site.
Review
Advances in the treatment of wound bacterial infection with phage
Cui Zelong
2015, 31(5): 389-391. doi: 10.3760/cma.j.issn.1009-2587.2015.05.019
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Abstract:
The treatment of wound bacterial infection is an extremely difficult problem in clinic, especially in patients with large wounds which are infected by multidrug resistant, pan-resistant or omni-resistant bacteria. In recent years, with a grim prospect of antibiotic resistance, phage therapy is re-valued by researchers after being ignored for nearly half a century. Phage therapy has made great achievements in prevention and control of bacterial infection of open wounds. This review is mainly focused on the latest research progress of phage therapy in wound bacterial infection.

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