BACKGROUND: Under special conditions, bone marrow mesenchymal stem cells (BMSCs) can differentiate into osteoblasts and chondroblasts. However, MSCs are few in bone marrow. How to harvest, purity and rapidly proliferate in vitro is a foundation of application in tissue engineering technique. OBJECTIVE: To optimize, collect, purity, assess rabbit BMSCs and to observe the biological character of BMSCs. DESIGN, TIME AND SETTING: The observational study was performed at the Animal Experimental Center of Tongji Medical College from September 2005 to July 2006. MATERIALS: One female New Zealand rabbits aged 2 months were used for MSC collection and primary culture. METHODS: Bone marrow solution was purified by density gradient centrifugation and adherence screening method. Culture solution was obtained. BMSCs were incubated in phosphate buffered solution (PBS), supplemented with 2.5 g/L trypsin (3.0 mL), and placed in an incubator at 37 ℃ for two or three minutes. Cell morphology was observed using an inverted microscope. The digestion was stopped when cytoplasm recovery, long and thin cells with large intercellular space, and few round cells appeared. Subsequently, BMSCs were incubated in serum-free L-DMEM, and placed in a plastic culture flask at 1.0×108/L. MAIN OUTCOME MEASURES: MSC morphology, ultrastructura and surface marker; Proliferation of the first, third, fifth, eighth and tenth passages of BMSCs; Cell growth curve was drawn. RESULTS: BMSCs was pure following density gradient centrifugation and adherence screening method. The third and fifth passage of cells had typical whirlpool-shape. Transmission electron microscope demonstrated that round or oval MSCs possessed large nuclei, big nucleus proportion, a few cellular organ. These were low-differentiated cells. Growth curve of cultured MSCs was "S" shape. The first, third and fifth passage cells had strong reproductive capability. The eighth and tenth passage of cells had significantly reduced proliferation. Cells isolated were positive for CD44 and CD90, but negative for CD34. These were low-differentiated cells under the electron microscope. CONCLUSION: Isolated cells are MSCs, with the property of stem cells. The third and fifth passage cells are pure, with strong reproductive capability.

Objective To investigate the effect of fat-1 gene encoding n-3 fatty acid desaturase on the proliferation and apoptosis of colon cancer cell HT-29. Methods fat-1 gene was transfected into HT-29 cells by liposomal reagent. The expression of fat-1 gene was detected by fluorescent micrographs and RT-PCR. Gas chromatography, MTT and flow cytometry were used to examine the change in n-6/n-3 PUFAs ratio, proliferation, cell cycle and apoptosis, respectively. Results After transfection of fat-1 gene, n-6/n-3 PUFAs ratio decreased significantly. Apoptosis of HT-29 cells was induced and cell cycle was changed. Apoptosis mainly appeared in the synthesis phase. Conclusion fat-1 gene encoding n-3 fatty acid desaturase can significantly decrease n-6/n-3 ratio. Consequently, apoptosis was triggered and cell cycle was changed. Tranfection of fat-1 gene into HT-29 cells may be a new potential treatment for colon cancer.

Objective To explore the clinical features and feasibility of renal transplantation for end-stage renal disease (ESRD) following hematopoietic stem cell transplantation (HSCT).Method A retrospective study was done in one case of renal transplantation for ERSD following HSCT.Clinical manifestations were summarized and prognosis was described.The 22-year-old male recipient had received HLA allele matched related bone marrow transplantation from his sister in 2001 and accepted renal transplantation 14 years after HSCT because of delayed renal dysfunction.Donor was a cardiac death patient,the preoperative Panel Reactive Antibody Testing was negative and there were 1.5 HLA antigen mismatches of 6 HLA-A,B,DR antigens of donor and recipient.The recipient received immunosuppressive therapy of tacrolimus + mycophenolate mofetil + steroid after renal transplantation.Result The patient's renal function remained stable and serum creatinine level was 65 μmol/L.The outcome of the patient was fairly good during the follow-up period of short-term.Conclusion Renal transplant is a feasible alternative for patients with ESRD following HSCT.If the transplanted kidney and abbr.hematopoietic stem cells are from different donors,irnmunosuppressive treatment is essential after renal transplantation.Long-term follow-up and adjustment of immunosuppression treatment are needed to prevent and treat postoperative complications.

OBJECTIVETo explore the impact of killer cell immunoglobulin-like receptor (KIR) gene mismatch on the outcomes of renal transplantation.

METHODSWe collected the data from 111 donor-recipient pairs of kidney transplant and analyzed the status of KIR gene matching, acute rejection (AR), and 1-year and 3-year survival of the recipients who were followed continuously for over 37 months.

RESULTSSeventeen KIR genes were expressed in both recipient and donor groups, and the frequency of KIR3DS1 was significantly higher in the recipients than in the donors (38.75% vs 24.66%, OR=2.17, χ² = 3.94, P<0.05). The average rate of donor-recipient KIR matching was 82.53%. The donor-recipient KIR2DS1 matching rate was significantly higher in AR group than in no-AR group (85.00% vs 54.95%, χ² = 6.19, P<0.05). The rate of donor-recipient KIR AB-AB genotype was significantly higher in AR group than in no-AR group (33.33% vs 8.00%, P<0.05). The 1- and 3-year survival rates was 94.59% and 82.88% in these recipients, respectively. The frequency of donor KIR-AB genotpye was significantly higher in recipients with poor outcomes (57.89% vs 29.63%, χ² = 8.19, P<0.05); the frequency of both donor and recipient KIR-AB genotype was also significantly higher in recipients with poor prognoses (36.84% vs 9.78%, χ² = 14.87, P<0.05).

CONCLUSIONSKIR3DS1 may be the susceptible gene associated with uremia. A KIR-AB genotype of either the donor or the recipient can increase the risk of AR and reduce the 1- and 3-year survival rate. This finding can be of ethically importance in choosing a living related donor.

Genotype ; Humans ; Kidney Transplantation ; Receptors, KIR ; genetics ; Survival Rate ; Tissue Donors ; Treatment Outcome

Objective To investigate the association between serum betatrophin levels and insulin resistance in newly diagnosed type 2 diabetes patients. Methods A total of 31 healthy subjects (control group) and 35 newly-diagnosed type 2 diabetes patients(new T2DM group) were recruited. Serum betatrophin was detected with the methods of ELISA, insulin resistance was assessed with homeostasis model(HOMA-IR). Results The levels of betatrophin, free fatty acids, HOMA-IR, fasting plasma glucose, fasting serum insulin, high-density lipoprotein cholesterol and low-density lipoprotein cholesterol were significantly higher in group B than those in group A (P<0.05). Conclusion Insulin resis-tance leads to elevated serum levels of betatrophin in patients with newly diagnosed type 2 diabetes.

ObjectiveTo evaluate the clinical efficacy and safety of Tripterygium wilfordii polyglycoside tablets （TWP） in the treatment of rheumatoid arthritis （RA） in the real world. MethodDiagnosis and treatment data of patients with RA in Chinese medicine rheumatology registration research information platform information database （CERTAIN） from January 1，2019 to January， 2024 were collected. According to the inclusion and exclusion criteria， data were screened. The included data were divided into an exposure group and a control group according to the use of TWP or not. Propensity score matching （PSM） was used in both groups to keep the baseline balanced and comparable. The disease activity score （DAS28-ESR） of 28 joints based on the erythrocyte sedimentation rate （ESR）before and after treatment was compared between the two groups， as well as health assessment questionnaire （HAQ），visual analogue scale （VAS），tender joint count （TJC），swollen joint count （SJC）， patient's global assessment （PGA），evaluator's global assessment （EGA），laboratory indexes， clinical curative effect， and adverse reactions. ResultA total of 3 978 patients were included，including 929 in the exposure group and 3 049 in the control group. Before PSM，there were significant differences in demographic information，DAS28-ESR score，PGA，EGA，HAQ，VAS scores，SJC， and TJC between the two groups （P<0.05）. After successful PSM matching，922 patients in the exposure group and 922 patients in the control group were included. There was no significant difference in demographic information and DAS28-ESR between the two groups before treatment，and the differences in other indexes between the two groups decreased to varying degrees. After treatment，the DAS28-ESR，PGA，EGA，HAQ，SJC，TJC，VAS scores， ESR，and IgG immune index of the two groups were significantly lower （P<0.01）. Compared with those in the control group after treatment，the DAS28-ESR，PGA，EGA，HAQ，VAS scores， and ESR in the exposure group after treatment decreased more significantly （P<0.05，P<0.01）. There was no significant difference in TJC in the exposure group after treatment. However， TJC in the exposure group was significantly higher than that in the control group before treatment （P<0.05）. In terms of TJC reduction，the exposure group performed better than the control group. There was no significant difference in SJC and IgG between the exposure group and the control group after treatment. After treatment，the clinical symptoms of poor appetite，insomnia and many dreams，upset，fatigue，and fear of wind and cold in the two groups were improved. Except that the proportion of women in the exposure group was higher than that in the control group （P<0.01），there was no significant difference in the incidence of other adverse reactions between the two groups after treatment. ConclusionTWP to treat RA can effectively reduce DAS28-ESR，PGA，EGA，HAQ，TJC，and VAS scores and improve the general symptoms. Except for the women at childbearing age with fertility requirements that TWP is not applicable，it shows good security.