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.

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.

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.

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.

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.

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.

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.

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.

Objective:To explore the anatomical characteristics of the anterolateral thigh perforator flap pedicled with oblique branch trunk of lateral circumflex femoral artery in the muscular septum and the clinical effect of this flap in repairing skin and soft tissue defects in the extremities.Methods:A retrospective observational study was conducted. From December 2020 to April 2021, 59 patients with skin and soft tissue defects in the extremities admitted to the Department of Hand Surgery of Suzhou Ruihua Orthopaedic Hospital met the inclusion criteria, including 46 males and 13 females, aged 20 to 81 years. The wound area after debridement ranged from 8 cm×5 cm to 38 cm×20 cm. According to the condition of the wound, the perforators in the anterolateral femoral region on the unaffected side were located by color Doppler ultrasound. The anterolateral thigh perforator flap with the oblique branch of lateral circumflex femoral artery as the source artery was designed, and the wound was repaired by unilateral flap or series combination of bilateral flaps (with the area of unilateral flap ranging from 7 cm×5 cm to 37 cm×11 cm). The wound of the donor site was sutured directly. The following items were recorded, including the number of perforators in the anterolateral femoral region marked before operation, the course characteristics of oblique branch trunk of lateral circumflex femoral artery and its perforators, and the number, origin, and type of perforators observed during operation, the flap repair mode and the flap harvest time. After operation, the survival condition of the flap, the wound healing time in the recipient site, and the suture healing in the donor area were observed, and the recovery of the donor and recipient areas was followed up. At the last follow-up, the sensation function evaluated by sensory rating scale of British Medical Research Association and two-point discrimination of the area transplanted with flap were recorded, and the improved comprehensive curative effect evaluation scale of flap was used to evaluate the repair effect of the flap.Results:A total of 156 perforators were marked in the anterolateral femoral region before operation, and 144 perforators were observed during the operation, of which 98 came from the oblique branch of the lateral circumflex femoral artery, and the first perforator of the oblique branch was the intermuscular septal perforator. Once formed, most oblique branch trunk of lateral circumflex femoral artery in the muscular septum could be divided into the deep branch and the superficial branch at the middle and upper 1/3 junction of the line between the anterior superior iliac spine and the lateral edge of the patella, the deep branch mainly ran in the muscle of vastus intermedius and vastus lateralis, and rarely developed the skin perforators, while the superficial branch mainly ran in the muscular septum between the rectus femoris and the vastus lateralis, and grew out the perforators to the proximal skin of the anterolateral femoral region. Fifty-six patients were repaired with unilateral flap and 3 patients with bilateral flap in series combination, with the harvesting time of the unilateral flap ranged from 9 to 99 min. Three patients developed arterial crisis within 48 hours after operation and survived after timely exploration; the flap of 1 patient developed necrosis 11 days after operation and was repaired by abdominal split-thickness skin graft; the other flaps survived smoothly. The wound healing time in the recipient area was 10 to 42 days after operation, and the sutures in the donor area healed well. During the follow-up of 6 to 10 months, 8 patients underwent thinning and plastic surgery 6 months after operation because of bloating in the flap transplantation area, while the other patients had a good shape of the flap transplantation area, with no deep tissue infection such as osteomyelitis, with soft texture, good elasticity, no pain, and good blood circulation; all the donor areas were left with linear scars, normal blood circulation in the distal extremities, no limitation of knee joint motion or quadriceps muscle strength. At the last follow-up, the sensation function was restored in the flap transplantation area of the affected limb, including 14 cases of S 1 grade and 45 cases of S 2 grade; there was only one point of two-point discrimination; the curative effect evaluation of flap repair included excellent in 24 cases, good in 35 cases, and general in 3 cases. Conclusions:The anterolateral thigh perforator flap pedicled with oblique branch trunk of lateral circumflex femoral artery in the muscular septum has the advantages of relatively concealed donor area, large cutting width, and simple operation during surgery. It protects the donor area to the greatest extent while repairing skin and soft tissue defects in the extremities, and is a useful supplement to the classic anterolateral thigh perforator flap.

Objective:To investigate the curative effects of the superficial peroneal artery perforator flap carrying two and more homologous perforators in repairing hand and foot wounds.Methods:A retrospective observational study was conducted. From January to September 2021, 23 patients with hand and foot wounds combined with bone or tendon exposure who met the inclusion criteria were admitted to Suzhou Ruihua Orthopaedic Hospital, including 16 males and 7 females, aged 23 to 68 years. After expansion of the wound, the wound area was 3.0 cm×2.0 cm to 11.0 cm×4.0 cm. All the wounds were repaired with the superficial peroneal artery perforator flap carrying two and more homologous perforators. The area of the resected flap was 4.0 cm×2.0 cm to 12.0 cm×5.0 cm. All the wounds in donor areas were sutured directly. During the operation, the resection of the flap, the number and caliber of carried perforators, the caliber of superficial peroneal artery, the length of vascular pedicle, and the number of accompanying veins of the superficial peroneal artery were recorded. The survival of the flap, the occurrence of complications, and the wound healing in the donor area were observed after operation. The recovery of the donor and recipient areas was followed up. At the last follow-up, the comprehensive evaluation scale was used to evaluate the curative effect of flap repair, and the sensory grading scale was used to evaluate the sensory function of the recipient area.Results:Totally 24 flaps were successfully resected in surgical operations, carrying 56 superficial peroneal artery perforators in total, with the caliber of perforators of 0.20-0.70 mm. The calibers of all perforators carried by 7 flaps were smaller than 0.40 mm. Nineteen flaps carried 2 perforators each, 3 flaps carried 3 perforators each, 1 flap carried 4 perforators, and 1 flap carried 5 perforators. During the operation, the superficial peroneal artery was resected with a caliber of 0.40-1.50 mm, the vascular pedicle was 2-6 cm in length, and each superficial peroneal artery had two accompanying veins. After operation, all the flaps survived smoothly, no vascular crisis or distal necrosis occurred, and the wounds healed well in the donor area. During the follow-up of 6 to 10 months after operation, the color, texture, and elasticity of the recipient area were good. Among them, 6 recipient areas were thinned and reshaped because of bloating. There was only linear scar and no obvious scar hyperplasia or pigmentation in the donor area, without significant change in sensory or motor function. At the last follow-up, the curative effect evaluation of flap repair was excellent in 22 flaps and good in 2 flaps, and the sensory function evaluation of the recipient area was grade S 3 in 1 area and grade S 2 in 23 areas. Conclusions:The superficial peroneal artery perforator flap with two and more homologous perforators has sufficient and reliable blood supply and is effective in repairing hand and foot wounds. It provides an ideal solution for the clinical problem in which the original operation scheme is abandoned due to the existence of only multiple slender perforators of caliber smaller than 0.40 mm, and only a single perforator in the operative field that cannot satisfy the needs of the flap blood supply and recipient area.