1.The feasibility study of repairment of articular cartilage defects with bone marrow-derived mesenchymal stem cells seeded on acellular amniotic membrane
Hao CHEN ; Zhijin ZHANG ; Dewei ZHAO ; Lin GUO
Chinese Journal of Microsurgery 2014;37(3):254-257
Objective To study the effect of repairment of articular cartilage defects in non weight-bearing area of rabbit with bone marrow-derived mesenchymal stem cells (BMSCs) seeded on human acellular amniotic membrane (HAAM).Methods From July 2012 to March 2013,bone marrow-derived mesenchymal stem cells were isolated and purified from rabbit in vitro.The cells were seeded on human acellular anniotic membrane at the concentration of 1.63 × l05/cm2.From 7 days to 8 days after cultured,the complexes of BMSC and HAAM were examined under electronmicroscope,light microscope and by HE stain.Full thickness empty defects measuring 4 mm in diameter by 3 mm depth were prepared in femoral intercondylar fossa of 24 rabbits.The rabbits were randomized into two groups:group A and group B with 12 each group.The defects of right knees were served as control and the left as experimental group.BMSCs/HAAM composite was cultured and then transplanted into the defect of left knee joint in group A as group BMSCs/HAAM and HAAM into group B as group HAAM.These rabbits were killed at 4 and 12 weeks after surgery in each group and the newly cartilage samples were evaluated grossly,histologically are graded.Results In the 4th and 12th week after the operation,the regenerated tissue were white,soft and smooth.Chondrocytes were found in the tissue In the 12th week,the morphology,distribution and arrangement of the regenerated tissues were similar to normal cartilage in the knees with HAAM-BMSCs transplantation.The regenerated tissues grew to be integrated with the surrounding normal cartilage with obscure boundary between them.Chondrocytes were found in all layer of the tissue,surrounding normal cartilage with obscure boundary between them.In the HAAM transplantation,the rough surface of regenerated tissue sunk obviously and the fibmblasts in all layer were found.While there were no regenerated tissue in the control side.Conclusion BMSCs seeded on HAAM could repair the articular cartilage defects of femoral intercondylar fossa from rabbits.
2.Identification and characterization of the integrated site of pseudomonas aeruginosa phage PaP3
Lin WANG ; Xiancai RAO ; Fuquan HU ; Zhijin CHEN ; Yinling TAN
Journal of Medical Postgraduates 2003;0(05):-
Objective: Identification of the attachment site of phage PaP3 within the genome of Pseudo-monas aeruginosa PAS. Methods:The full genome of lysogenic bacteria was cleaved by Pst Ⅰ and produce a large fragment of more than 45 000 bp, which was subsequently digested by EcoR Ⅰ. Then the fragment containing DNA sequence of phage and bacteria was cloned into pFastBacTMHT A vector, and the result of sequencing indicated the right hybrid site attR. AttL was isolated by PCR on the base of integration mechanism. And then attP and attB were indentified according to the nucleotide sequences of attR and attB. Results:A sequence of 21 bp(5'-GGTCGTAGGTTCGAATCCTAC-3') was defined to be the core site of integration, which was located at t-RNAPro gene in the genome of phage PaP3 and t-RNALys gene in the genome of Pseudomonas aeruginosa PA3. The attP and attB flanked with a set of inverted repeat and direct repeat. Conclusion:The integrated site of PaP3 within the genome of PA3 was identified and characteriged, which could be of value in investigating the mechanism of integration and gene flow between different species in the natural world.
3.Metal implants for treatment of unstable pelvic fracture
Zhijin LIN ; Liehu CAO ; Feng SHEN ; Jianming HUANG ; Yifan KANG
Chinese Journal of Tissue Engineering Research 2010;14(9):1665-1668
BACKGROUND:It is controversial to treat sacroiliac joint fracture.Some scholars advocated expectant treatment,and some others advocated surgery therapy.Pelvic stability was responsible for the scheme selection.Sacroiliac joint fracture-dislocation destroys pelvic stability,which easily induces instability and bone nonunion,resulting in sacroiliac joint pain,unequal size of lower limbs,sitting pain and dysfunction.Thus,prognosis of mental implant is significantly better than expectant treatment in treatment of unstable pelvic fracture.OBJECTIVE:To summarize the treatment of unstable pelvic fractures and their clinical application using various internal and external fixation of metal implants.METHODS:The computer-based research was done in Pubmed database (http://www.ncbi.nlm.nih.gov/PubMed) and Wanfang database (http://www.wanfangdata.com.cn) for articles published from January 1991 to December 2009 with the key words of "Pelvic fractures,instability,surgical treatment" by the first author.A total of 115 articles were retrieved,and those concerning characteristics and clinical application of implants in the treatment of unstable pelvic fracture.Articles addressing old and repetitive contents were excluded.Literatures of the same fields published in recent years or in authorized journals were selected.Finally,30 articles were included.RESULTS AND CONCLUSION:Sacroiliac joint fracture-dislocation is a severe,high-energy trauma,has been paid great attention in the therapy,particularly in unstable sacroiliac joint fracture-dislocation.A stable type of fracture and dislocation of the sacroiliac joint received a conservative treatment of unstable sacroiliac joint fracture-dislocation appropriate line of external fixation,internal fixation for reconstruction of pelvic stability and internal fixation treatment varied,but the therapeutic effect of internal fixation needs to be improved.What are bio-mechanical characteristics of various internal fixation methods,and how the timing of weight-bearing activities following various internal fixations require further basic and clinical studies.An unstable sacroiliac joint fracture-dislocation fixation has many ways,including the anterior and posterior fixed-fixed.Minim ally invasive therapy such as posterior CT guided sacroiliac joint lag screw is the developmental trend.
4.Effect of low-intensity pulsed ultrasound on cell-free demineralized bone matrix co-cultured with rabbit cartilage cells and bone marrow mesenchymal stem cells in vitro
Zhijin LIN ; Hao TANG ; Feng SHEN ; Jianming HUANG ; Yifan KANG
Chinese Journal of Tissue Engineering Research 2009;13(47):9217-9223
BACKGROUND: Using low-intensity pulsed ultrasound (LIPU) to promote the repair of articular cartilage injury is very common,and we also have more options to choose the cytoskeleton, but the application conditions of LIPU and the appropriate cytoskeleton have not reached any consensus yet.OBJECTIVE: To investigate the feasibility of establishing tissue-engineered cartilage by cell-free allograft demineralized bone matrix (CFDBM) co-cultured with rabbit cartilage cells and bone marrow mesenchymal stem cells (BMSCs) in vitro, and to investigate the effect of LIPU on the cells in CFDBM.DESIGN, TIME AND SETTING: Multiple sample observation was performed at the Institute of Biomedical Engineering, Second Military Medical University of Chinese PLAfrom May to August 2009.MATERIALS: The CFDBM was prepared as modified Urist's method; the cartilage cells were obtained using mechanical disintegration and enzyme digestion; the BMSCs were separated using whole bone marrow rinsing method, purified, and amplified layer by layer.METHODS: As CFDBM With a composite of different cellular components, and whether applying LIPU stimulation, the samples were divided into four groups: chondrocyte group, BMSCs group, compound group (CFDBM was compounded with chondrocytes,BMSCs, and chondrocytes/BMSCs, respectively, without LIPU stimulation), and LIPU group (CFDBM was compounded with chondrocytes/BMSCs, and then the samples were stimulated with LIPU on the second day, 1.0 MHz frequency, 10 mW/cm~2 transient spatial intension, 20 min/d).MAIN OUTCOME MEASURES: ① the 2~(nd)-generation of cartilage cells and BMSCs were examined by immunohistochemical method; ② The CFDBM prepared as modified Urist's method was examined as HE staining; ③ The samples of four groups were examined by collagen II immunohistochemical staining on the 21~(st) day.RESULTS: ① The collagen II immunohistochemical staining of the second generation of the articular cartilage cells showed that the morphostructure was polygon, star or round, and pseudopodia extended, and the cells were rich in cytoplasm; the cytoplasm was brownish yellow, and the cell nuclear was round. ② The result of immunohistochemical staining of BMSCs showed that,CD34 was negative, CD44 and CD105 were positive. ③ In the center of CFDBM prepared as modified Urist's method, there was no obvious cell-like structure and the gap size was uniform. ④ On the 21~(st) day after combining CFDBM with cells, collagen II immunohistochemical staining demonstrated that BMSCs group was negative, chondrocyte group was weak positive, compoundgroup was positive, and the LIPU group was strongly positive.CONCLUSION: ① Biological property of the 1~(st)-3~(rd)-passage chondrocytes and BMSCs was similar to primary-cultured cells. ②Both chondrocytes and BMSCs had a highly proliferative ability in CFDBM. ③ 10 mW/cm~2 LIPU could not affect activity of BMSCs but could promote differentiation Into articular cartilage cells, and it also could not promote celt proliferation.
5.Randomised clinical trial on rh-Endo combined with FOLFOX4 regimen as an adjuvant therapy for stage Ⅱ and Ⅱ colorectal cancer patients
Zhihua XIE ; Dejian DAI ; Lin ZHONG ; Yi YI ; Jun FU ; Zhijin ZHANG ; Yuhao ZHANG
Chinese Journal of General Surgery 2013;28(10):758-762
Objective To evaluate the long-term efficacy of recombinant human endostatin (rh-Endo) combined with FOLFOX4 as an adjuvant treatment for patients of stage Ⅱ and Ⅲ colorectal cancer.Methods Eligible patients were randomly assigned to receive FOLFOX4 or FOLFOX4 plus rh-Endo regimen in which patients receiving 7.5 mg/m2 Ⅳ on day 1-7,repeated every 2 weeks,to a total of 12 cycles in 6 months.Results A total of 197 eligible patients were accrued in this research with 105 patients in the control group and 92 patients in the experimental arm.Median follow-up period was 42 months.The baseline characteristics distributed were balanced by treatment.Rh-Endo combined with FOLFOX4 regimen resulted in significant improvement on DFS compared to FOLFOX4 regimen for patients with stage Ⅲ colon cancer (HR =0.19,95% CI0.05-0.75,P =0.0124),and with a 34% improvement on 3-year DFS and 81% reduced recurrence.Although rh-Endo combined with FOLFOX4 regimen failed to make significant difference on DFS in the whole (HR =0.75,95% CI 0.31-1.83,P =0.5589),it was also observed a 17% improveiment on 3-year DFS.No statistical significant difference on DFS was observed in patients with stage Ⅱ disease.Conclusions Rh-Endo combined with FOLFOX4 regimen significantly improved the disease-free survival for patients with stage Ⅲ colorectal cancer,indicating that patients with stage Ⅲ disease,but not stage Ⅱ disease,can benefit from FOLFOX4 plus rh-Endo regimen in adjuvant treatment.
6.Investigation on epidemic foci of paragonimiasis in eastern part of Zhenghe County,Fujian Province
Benxiang LIN ; Huanwang WEI ; Yousong LI ; Shiqing WU ; Fanzhen KONG ; Zhijin FAN ; Baogui TANG ; Cai CHEN
Chinese Journal of Schistosomiasis Control 2016;28(4):418-421
Objective To investigate the species and hosts of Paragonimus and its infection rate in eastern part of Zhenghe County,Fujian Province,so as to determine the local foci of Paragonimus. Methods The snails,crabs and stools of wild cats were collected for the examinations of cercariae,metacercariae and eggs of Paragonimus. The geographical and environmental conditions of the areas were also investigated. Results A total of 4 890 Pseudobythinella jianouensis snails and 1 035 Semisul?cospira liberlina snails were examined,and the cercariae of Paragonimus were only found in P. jianouensis,with an infection rate of 0.10%(5/4 890). Bottapotamon zhengheensis sp. nov. as the second intermediate host of P. skrjabini,were examined, and the infection rate was 85.29%(29/34)and the average numbers of metacercariae per crab and per gram of crab tissues were 3.85 and 0.62,respectively. Thirty?six Sinopotamun fujianensis crabs,as the second intermediate host of P. westermani,were examined,and the infection rate was 38.89%(14/36)and the average numbers of metacercariae per crab and per gram of crab tissues were 6.43 and 0.03,respectively. The eggs of Paragonimus were detected in 1 of 2 muck specimens of wild cats. Conclu?sion The data suggest that there is a focus of middle?to?high level of infection caused by P. westermani and P. skrjabini in the eastern part of Zhenghe County.
7.Bilobed anterolateral thigh perforator flap pidicled with the oblique branch of lateral circumflex femoral artery in repair of the wound in extremity
Zhijin LIU ; Jihui JU ; Junnan CHENG ; Lin YANG ; Shengzhe LIU ; Tao ZHANG ; Ruixing HOU
Chinese Journal of Microsurgery 2021;44(2):152-156
Objective:To investigate the feasibility and clinical efficacy of the bilobed anterolateral femoral perforator flap pidicled with the oblique branch of lateral femoral circumflex artery in repair of the wounds in extremities.Methods:A study was conducted on 36 patients with complex limb wounds from December, 2014 to May, 2020. Thirty patients had single-wound sized from 10.0 cm×10.0 cm to 23.0 cm×17.0 cm, and 6 patients had 2 adjacent and discontinuous wounds sized from 4.0 cm×5.0 cm to 11.0 cm×9.0 cm. A total of 78 perforators were identified in routine preoperative CDU examination, and 67 perforators were confirmed. According to the actual requirement of the perforator confirmed in surgery, 19 flaps were designed with completely split up into bilobed flaps and 17 bilobed fascial flaps were designed with segmented skin and deep fascia. The blood supply of flaps relies on the anastomosis of oblique branch with the recipient vessels, therefore bridged blood Flow-through anastomosis was performed in 8 flaps. All thigh donor sites were sutured directly. Regular follow-up were made after surgery.Results:In this group, 35 cases of bilobed flaps survived successfully. Venous crisis was found in 1 case of flap repairing 2 wounds after the operation and was relieved 7 days later by remove some stitches and bloodletting. The donor sites healed by first intension, and the wound healing time was 11-83 days. All patients were followed-up for 6-39 months. The flaps had good colour and texture with S 2-S 3 sensory. All the donor sites left linear scars except 1 where left with a large scar without contracture and pain. Conclusion:The repair of the wounds in extremities by bilobed anterolateral thigh perforator flaps pedicled with the oblique branch of the lateral femoral circumflex artery could obtain a more concealed donor site. It acts as a beneficial supplement when a bilobed flap cannot be harvested on the descending branch of the lateral femoral circumflex artery.
8.Finite element comparative study of S2 alar-iliac screw and sacroiliac joint screw in treatment of type C sacroiliac joint dislocation
Huan MENG ; Guanghu LIN ; Xiaoreng FENG ; Fei LIU ; Zhijin CUI ; Bin CHEN
Chinese Journal of Trauma 2018;34(6):505-512
Objective To compare the biomechanical characteristics between S2 alar-iliac (S2AI) screw and sacroiliac joint screw in treating unilateral type C sacroiliac joint dislocation. Methods A 25 year-old healthy male adult agreed to take computed tomography scan of the pelvis. The pelvic floor thickness was 1 mm and the image data were saved in DCOM format. A three dimensional finite element model of intact pelvis was reconstructed by Mimics, Geomagic, SolidWorks, and Abaqus softwares. A finite element model of type C unilateral sacroiliac dislocation was established. One S1 vertebral sacroiliac screw of 6.5 mm in diameter (S1 group), one S2AI screw (S2AI group) and one Si vertebra sacroiliac screw + one S2 vertebra sacroiliac screw (S1+S2 group) were placed in the model, respectively. The500 N vertical load on the upper surface of the sacrum was simulated. The displacement value of sacroiliac joint, the displacement of sacrum, the stress value of the internal fixation and surrounding bone, and the stress distribution cloud map were recorded and analyzed. Results The displacement data showed that displacement distribution in the three groups had a consistent trend. In terms of the absolute value of displacement the result was S, group>S 2AI group>S1+ S2 group, of which the maximum displacement of sacroiliac joint in S1 groups was 0.87 mm and that of sacrum was 1.6 mm; the maximum displacement of sacroiliac joint in S2 AI group was 0.22 mm, and that of sacrum was 0.24 mm; the maximum displacement of the sacroiliac joint in S1 + S2 group was 0.06 mm, and that of sacrum was0.16 mm. Stress data showed that the stress in the internal fixation was mainly distributed at the sacroiliac joint, and the stress result was S2AI group>S, group>S, + S2 group, of which the maximum stress value was 52.8 MPa in S1 group, 62.1 MPa in S2AI group, and 38.2 MPa in S1 + S2 group. The stress around the screws was also concentrated at the sacroiliac joint, and the stress result was S1 group>S2AI group>S1 + S2 group, of which the maximum stress value was 56.8 MPa in S1 group, 11.2 MPa in S2AI group, and 5.8 MPa in S1 + S2 group. Conclusions Single S1 screw, single S2 AI screw and S, screws combined S2 screws can be used for the treatment of unilateral C type sacroiliac joint dislocation. Early weight bearing of single S1 screw might lead to the risks of increased sacroiliac joint displacement and internal fixation failure. S2 AI screw fixation and S1 + S2 screw fixation have similar biomechanical strength, allowing early weight-bearing and contributing to better postoperative rehabilitation.
9.Application of "ABC" three line perforator locating method for free anterolateral perforator flap of calf
Tao ZHANG ; Lin YANG ; Junnan CHENG ; Shengzhe LIU ; Zhijin LIU ; Yongtao HUANG ; Qinfeng GAO ; Fengwen SUN ; Chengpeng YANG ; Jihui JU
Chinese Journal of Microsurgery 2023;46(1):70-75
Objective:To explore the feasibility of an "ABC" three line perforator locating method in design and harvest of free anterolateral perforator flap of calf.Methods:Between March 2021 and November 2021, 42 patients with 62 wounds on hand and foot were treated in the Department of Hand Surgery, Suzhou Ruihua Orthopaedic Hospital. The "ABC" three line perforator locating method was applied to determine the location and source of perforating branch before operation and to guide the design and harvest of flap during operation in wound reconstruction. Among the 42 patients, 24 had the injury of single digit, 7 had the injuries with 2 digits, 4 with 3 digits, 1 with 4 digits, 1 of the first web, 1 in the wrist, 2 of the great toe, 1 of second toe and 1 in dorsal foot. The sizes of soft tissue defect were 1.5 cm×2.0 cm-3.0 cm×14.0 cm. The sizes of the flaps were 2.0 cm×2.5 cm-3.5 cm×15.0 cm. All donor sites were sutured directly. In the follow-up, sensations of flaps were evaluated following the sensory function evaluation standard of British Medical Research Council(BMRC), and the recovery of the donor and recipient sites was evaluated by the flap comprehensive evaluation scale. Regular follow-up were scheduled at outpatient clinic.Results:A total of 162 perforators were located before operation. There were 95 perforating branches being explored in the operation, of which 5 patients had 1 extra perforating branch than that located before surgery. Seventy-six perforating branches were found consistent with preoperative localisation, with a coincidence rate of 84.4%(76/90). Sixty-four perforating branches were found consistent with the preoperative source with an accuracy rate of 84.2%(64/76). All the 62 flaps survived without a vascular compromise. Follow-up lasted for 6-10(mean 7.1) months. The colour and texture of the flaps were excellent. The flaps were thin and wear-resistant. The sensory function of the flaps was evaluated at S 1-S 3 by BMRC. Comprehensive evaluation scale of flap was excellent in 38 patients and good in 4 patients. Conclusion:"ABC" three line perforator locating method in design of free anterolateral calf flap is a feasible and an ideal auxiliary method in surgical practice. It combines anatomical knowledge, clinical experience and Doppler ultrasound localisation as well as accurately guides the location and source prediction of perforator before surgery.
10.DSA of the oblique branch of lateral circumflex femoral artery: A morphological study and clinical significance
Lin YANG ; Junnan CHENG ; Shengzhe LIU ; Zhijin LIU ; Tao ZHANG ; Lei LI ; Jihui JU
Chinese Journal of Microsurgery 2021;44(6):642-646
Objective:The morphological characteristics of the oblique branch of lateral circumflex femoral artery (LCFA) were observed by digital subtraction angiography (DSA) in order to provide imaging basis in the application of the ALTF pedicled with the oblique branch of LCFA.Methods:Between February, 2020 and December,2020, for the patients who were requested to repair the wound with ALTF, a DSA radiography was performed before operation. A total of 197 sides of selective DSA were analysed in 113 patients. The occurrence rate, origin and course of the oblique branch of LCFA were observed. Relationships between the oblique branch of LCFA and the upper cutaneous branch, descending branch and transverse branch were analysed. In addition, in order to verify the accuracy of conventional DSA data in describing the morphological characteristics of oblique branches, 10 sites of 10 patients were randomly selected to perform rotational DSA three-dimensional imaging.Results:Femoral artery, deep femoral artery, LCFA and the branches of LCFA were clearly identified on DSA images. The oblique branch appeared in 190 sites, with a occurence rate of 96%. Among them, 1 oblique branch originated from the femoral artery, 2 from the deep femoral artery, and other 187 from LCFA. The oblique branches were found in 10 sites from rotational DSA three-dimensional imaging, which was consistent with conventional DSA imaging.Conclusion:The occurrence rate and morphological characteristics of the oblique branch of LCFA can be directly analysed by DSA. The oblique branch is not a variant branch as reported in the literatures, as it always appears. It may serve the main blood supply artery of the anterolateral thigh flap.