BACKGROUND:Studies have reported that puerarin can reduce bone resorption and promote bone formation. OBJECTIVE: To further explore the effects of puerarin at different doses on the proliferation of osteoblasts culturedin vitro. METHODS: Osteoblasts isolated from the cranium of newborn rats were culturedin vitro, and then passage 3 cels were cultured in DMEM medium with the presence of 0, 20, 40, 80 and 100 μmol/L puerarin+10% fetal bovine serum for 48 hours. The viability of osteoblasts, alkaline phosphatase activity and mineral node formation were determined using MTT, alkaline phosphatase kit and alizarin red staining, respectively. RESULTS AND CONCLUSION:The viability of osteoblasts treated with puerarin at either 40 or 80 μmol/L was significantly higher than that of the control group. Alkaline phosphatase activity and the number of mineral modules were significantly increased in osteoblasts cultured with puerarin at 40 or 80 μmol/L when compared with the untreated cels. These findings demonstrate that puerarin is able to promote osteoblast proliferation in vitro.

Objective To assess the efficacy of laparoscopic salpingostomy in infertile women with distal tube obstruction. Methods 109 women with infertility underwent laparoscopic salpingostomy and the results were retrospectively analyzed. Results The total cumulative pregnancy rate was 32.9%.The cumulative pregnancy rate was 44.4% in the unilateral distal tube obstruction group after laparoscopic salpingostomy,while it was 20% in the bilateral distal tube obstruction patients(p

Objective To investigate the clinical value of the YSZ-1 uterine manipulator in total laparoscopic hysterectomy.Methods Total laparoscopic hysterectomy was employed in a total of 78 women by using the LigaSure,ultrasonic scalpel,and the YSZ-1 uterine manipulator.The anterior fornix was opened and the manipulator was placed on the lip of the cervix.Then the uterus was completely removed.Results The operation was completed smoothly in all the 78 patients.The operating time was 45~90 min(mean,63 min),and the intraoperative blood loss was 40~100 ml(mean,65 ml).No urinary tract injuries,vaginal bleeding,or other postoperative complications occurred.The length of postoperative hospital stay was 4~7 d(mean,5.3 d).Follow-up examinations in the 78 patients for 3~6 months(mean,4 months) showed that the vaginal wound normally healed up. Conclusions Use of the YSZ-1 uterine manipulator in total laparoscopic hysterectomy makes the procedure easier and safer.The YSZ-1 uterine manipulator is a useful laparoscopic instrument.

Objective To explore the potential applications of a chamber for in vivo tissue engineering,and to establish a novel model for in vivo tissue-engineered cartilage.Methods Auricular chondrocytes were isolated,cultured and identified from the ears cartilages of New Zealand white rab bits; rabbit auricular chondrocytes(RACs) were seeded into the scaffolds:(1) RACs were seeded into collagen gel scaffold; (2) RACs were seeded into PLGA/collagen gel scaffold in vitro,and the compos ites were placed into the chambers and implanted in the donor rabbit.As control groups,the composites were implanted directly subcutaneously in the donor rabbit without using chambers,and the contents were harvested at 8 weeks after implantation.Gross examination,histologic and immunohistochemical staining and RT-PCR test were performed to evaluate the harvested contents.Results Under the same conditions inside the chambers,the contents formed into new cartilage-like tissue by histo logical and immunohistochemical staining and RT-PCR.In contrast,in the control groups without chambers displayed vascular invasion and inflammatory reaction in the subcutaneous layer of skin,which eventually led to fibrous tissue or absorption.Conclusions Cartilage is successfully constructed in an immunocompetent animal model using a bioinert perforated chamber.This method is effective in creating a relatively favorable environment for cartilage regeneration,which may provide a valuable reference for the clinical application of tissue regeneration.

Objective To understand acute rejection differences between untreated recipients and rapmycin-treated recipients in a rat free flap allotransplantation model. Methods Brown groin free flaps were transplanted to Lewis recipients. In the treated group, recipients were treated with rapamycin at the dose of 4 mg/kg every day from day 0 to day 14 after transplantation. In the untreated group, recipients didn't receive any treatment. Allografts were evaluated clinically and histologically. Results Allografts in the treated group showed epidermolysis as sign of rejection.Rejection sign of untreated grafts was ischemic necrosis of whole skin. In histological evaluation, the treated grafts showed "band-like" lymphocytes infiltration in the upper dermis when rejection occurred, while the untreated grafts showed thrombosis in the subdermal vessels. Conclusion The differences between the two groups implied that embolization may be responsible for the rejection of free flap allotransplantation in rat model.

Objective To evaluate the safety and the feasibility of hypophysin injection for hemostasis during laparoscopic stripping of ovarian endometrioma.Methods Retrospective analysis of 86 cases with ovarian endometrionm.Forty-two patients with prophylactic dilute hypophysin injection into cornua uterus and mesosalpinx(study group),and 44 patients without vasopressin(control group).The operative time,intraoperative blood loss,the highest postoperative temperature,postoperative stay and the rate of recurrence were compared.Results The operative time in the study group and the control group were(51.24±22.58)min and(67.02±25.14)min,the intraoperative blood loss were(42.16±26.10)ml and (68.23±28.21)ml,respectively.There was significantly different between two groups(P＜0.01).The rates of recurrence in the study group and the control group were 4.76% and 11.36%,respectively.But it was no significantly different between two groups(P＞0.05).Both the highest postoperative temperature and postoperative stay was no difference between two groups(P＞0.05).Conclusions Hypophysin can reduce the operative time and intraoperative blood loss during laparoscopic stripping of ovarian endometrioma,and it is possible to reduce the rate of recurrence.It is a safe,feasible,cheapand convenient method,and worth using in clinic.

BACKGROUND: The functional gene fragments integrate into gene vector, which is then transfacted into target cells or joint cavity, through the transgenic target cells continue to secrete a large number of functional gene product, local therapeutic concentrations could be maintained within a long period of time, thus repairing articular cartilage injury. OBJECTIVE: To transfect rabbit articular chondrocytes using recombinant retroviral vector-mediated transforming growth factor-βl (TGFβ_1) in vitro, and to observe its expression and its effect on biological characters of chondrocytes. METHODS: Rabbit chondrocytas were isolated by use of trypsin digestion method. Vector was PLNCX_2 Hind Ⅲ/Not Ⅰ doubly digested and dephosphorylated, connected with some multiple cloning sites and RFP gene following pDsRed_2 double digestion, to build PLNCX_2-RFP. TGFβ_1 gene was amplified from the PGEMT-TGF and connected with PLNCX_2-RFP following double digestion, to build PLNCX_2-TGFβ_1-RFP. Subsequent to packaging retroviral vector, viral supernatant titer was detected. The cultured and transfected chondrocytes in rabbit knee joint were divided into 3 groups: control group (without any transfection), transfected PLNCX_2 group and transfected PLNCX_2-TGFβ_1-RFP group, continued screening 2 weeks to observe the cellular changes. Cell supematant transfected stably were collected for detecting the effect of gene transfection on the chondrocytes with NO detection kit, ELISA assay was applied to determine human TGFβ_1 expression in cell culture supernatant. RESULTS AND CONCLUSION: The recombinant gene PLNCX_2-TGFβ_1-RFP was identified correct sequence by the enzyme digestion sequencing TGFβ_1 and RFP, which showed that the eukaryotic expression vector PLNCX_2-TGFβ_1-RFP had been successfully built as expectation. They were then transfected into packaging calls and cultured, the virus titer was defined as 1×10~6 CFU. Following stable transfection of cartilage cells, red fluorescence can be observed, proving successful transfection. After continuous screening 2 weeks, the scattered adherent calls formed positive clones, and gradually diffusely integrated, cell clusters appeared with common dual cores, the calls proliferated actively. NO concentration in the transfected PLNCX_2-TGFβ_1-RFP group was higher than that of transfected PLNCX_2 group (P < 0.05), no difference was significant between control group and transfected PLNCX_2 group. The control group and the group transfected PLNCX_2 showed no TGFβ_1 expression, while TGFβ_1 concentration was (28.08±3.73) ng/L in the transfected PLNCX_2-TGFβ_1-RFP group. PLNCX_2 ratroviral vector-mediated human TGFβ_1 can be effectively transfected into rabbit knee joint cartilage cells and obtain stable expression, while the transfected cartilage calls proliferate actively.

Objective:To construct the siRNA expression vector of focal adhesion kinase(FAK) gene and inhibit the expression of FAK gene in tongue cancer cell line Tca8113 by RNA interfering technique. Methods:According to the encoding sequence of FAK mRNA, two pieces of oligonucleotide sequences were designed and synthesized. The annealed oligonucleotide fragments were subcloned into pGCSilencerTM-U6/Neo siRNA expression vector. After being identified by restriction enzyme method, the recombinant pSilencer-FAK plasmids were transfected into Tca8113 cells. The transfected cells were selected by G418 method. Immuocytochemistry and Western blotting were used to evaluate FAK gene silencing efficiency. Results:The oligonucleotide fragments were correctly inserted into pGCSilencerTM-U6/Neo vector. FAK expression of the transfected cells was significantly down-regulated by pSilencer-FAK. Conclusion:The siRNA expression vector of FAK is successfully constructed and FAK expression of Tca8113 cells can be inhibited by RNA interfering technique.

Objective To evaluate the efficacy and quality of life of segmental bowel resection for bowel endometriosis. Methods Totally 62 symptomatic patients with bowel endometriosis undergoing segmental bowel resection were recruited. A visual analogue scale (VAS) and the 36-item short form health survey (SF-36) questionnaire were administered before and at least 1 year after surgery, respectively. Pregnancy rates were also recorded. Results Sixty-two patients in total underwent follow-up ranging from 12 to 74 months. All patients complained of obvious pain symptoms, including dysmenorrhea, dyspareunia, pain on defecation and chronic pelvic pain. The relief of dysmenorrhea (2.9 ± 2.2 versus 7.5 ± 2.9), dyspareunia (0.7 ± 0.5 versus 4.3 ± 2.2) and pain on defecation (1.6 ± 0.7 versus 7.3 ± 1.9) after surgery was statistically significant (all P<0.01). The scores for all 8 domains of the SF-36 questionnaire were significant improved after segmental bowel resection (all P<0.01). The complication rate was 45% (28/62), including 18 cases of urinary retention, 4 rectovaginal fistulas, 2 cases of vaginal dehiscence, and 1 case each of thrombogenesis, pelvic abscess and general peritonitis. All of the patients with complications recovered well throughout follow-up. The postoperative pregnancy rate of the previous infertile patients was 6/10. Among the 6 gestational cases, 2 had labour, 2 underwent caesarean sections, one had a spontaneous natural abortion, and one underwent uterine curettage. Conclusion Segmental bowel resection could significantly relieve pain and improve quality of life for patients with bowel endometriosis.

Objective To investigate the effect of radiofrequency ablation(RFA) in the treatment of small liver cancer.Methods In recent 6 years,130 patients with primary or secondary liver cancer(≤5cm in diameter)were treated by RFA,among which 86 cases were primary,and 44 cases were secondary liver cancer.Among the 130 cases,18 received RFA plus TAEC,and 20 had RFA combined with PEI.Results Overall 1-and 3-year survival rate was 91.3% and 77.7% respectively.Complications occurred in 7 cases,including biliary fistula,intestinal fistula,and slight burn of skin.There was no death in this series.Conclusions RFA is an effective and safe therapeutic approach for small liver cancer ≤5cm in diameter.Its effect may be similar to that of surgical resection.