BACKGROUND:We attempt to explore a low-cost, simple and effective way to cryopreserve bone marrow mesenchymal stem cels at-80℃.
OBJECTIVE:To screen the optimal cryopreservation fluid for bone marrow mesenchymal stem cels and to verify the biological features of bone marrow mesenchymal stem cels after long-term cryopreservation.
METHODS: Bone marrow mesenchymal stem cels were cultured using adherent method and the biological features and purity of cels were detected using immunofluorescence method. Bone marrow mesenchymal stem cels were cryopreserved in the cryoprotectant medium containing low-sugar DMEM, fetal bovine serum and dimethyl sulfoxide at different proportions at-80℃ for a short term. Then, the optimal cryoprotectant was selected to storage the bone marrow mesenchymal stem cels. After 1, 3, 6 months of cryopreservation, the cels were resuscitated, cultured and passaged. Passage cels were identified immunofluorescence method to determine the biological features of bone marrow mesenchymal stem cels cryopreserved at-80℃.
RESULTS AND CONCLUSION:Cryoprotectant medium of 80% DMEM+10% fetal bovine serum+10% dimethyl sulfoxide was suitable for cryopreserving MSCs at -80℃, and resuscitated cels were able to proliferate in vitro, and passage normaly, indicating the cryopreserved bone marrow mesenchymal stem cels stil maintain the original biological activity.

Objective: To estimate the clinical efficacy of combination of chemotherapy with osteoblast promoting and anti-osteolys-is agents in bone metastatic carcinoma. Methods: Seventy-one patients with bone metastatic carcinoma were divided into 2groups and treated with protocol A or B, respectively. The protocol A was a combination of chemotherapy with osteoblastpromoting and anti-osteolysis agents, the protocol B was a combination of chemotherapy with antiosteolysis agent only. Theefficacies of the 2 protocols were compared and studied. Results:The effect of A was more remarkable than that of B in re-lieving bone pain and enhancing bone repair. No severe adverse effect of these agents was found in therapy. Conclusion: Theclinical efficacy of combination of chemotherapy with osteoblast promoting and anti-osteolysis agents in bone metastatic car-cinoma was definite, the osteoblast promoting agent as an effective adjtmctive agent can be used not only in osteoprosis butalso in bone metastatic carcinoma.

Objective: In order to reveal the mechanisms of rh-endostatin and its suppression effect on HUVECs at different activating levels. Methods: Three types of HUVEC were prepared. We detected the ex-pression of CD105 and CD62E on the surface of adhering HUVEC, HUVEC of exponential phase of growth, and HUVEC activated with TNF-alpha. The effect of endostatin on these three types of HUVEC was indicated by the variation in the expression of CD105 and CD62E on the cells. The relationship of the changes in cell markers with the time and the concentration of endostatin was analyzed by SPSS 11.5. Results: The expres-sion of CD105 was correlated with time and dose of endostar in all groups. The expression of CD62E was cor-related with the dose of endostar in HUVEC treated with TNF-α for 24h. Conclusion: Endostatin of a certain dose can inhibit the activation of endothelial cells but it cannot induce cell apoptosis.

Objective To survey the clinical effect of combination therapy of Ninggan-Tiaozhong decoction and psychological intervention for chronic atrophic gastritis (CAG).Methods 80 cases CAG were randomly divided into a treatment group (treated with Ninggan-Tiaozhong decoction and psychological intervention) and a control group (treated with Ninggan-Tiaozhong decoction),with 40 cases in each group and performed a therapy for 12 weeks.The clinical efficacy and the change of Symptom Checklist (SCL-90)and Self-Rating Anxiety Scale(SAS) before and after treatment were evaluated.Results After 12 weeks of treatment,the total effective rate was 90.0％ in the treatment group,and 74.4％ in the control group.The treatment group was better than the control group(x2=4.255,P＜0.05).After psychological intervention,SCL-90 and SAS score (39.5±4.3,1.3± 0.6)was significantly reduced in the treatment group than the control group (51.4±3.2,1.5±0.3),(P＜0.05).Conclusion The combination therapy of Ninggan-Tiaozhong decoction and psychological intervention used in CAG is effective to relieve symptoms and to improve life quality of patients.

Objective:To study the direct effects of gradient concentration inteiferon-a(IFN-a) on lymphoma cells line Daudi and Jur-kat and a group of 15 cases refractory lymphoma. Methods: The effects of IFN-a on the growth of tumor cells were assayed by MTT, apoptosis were measured by flow cytometty, TUNEL and electron-microscope, 15 refractory lymphoma cases were treated by local injection of IFN-a combination chemotherapy.Results: Low-dose IFN-a did not have any significant effects on growth of Daudi and Jurkat cells, high-dose IFN-a (10 000U/ml) not only have significant anti-proliferative effects with dependence of time and dosage but also can induce apoptosis on both Daudi and Jurkat cells. 5 patients achieved a complete remission and 7 patients achieved a partial remission, the overall response rate was 80% .Conclusion:IFN-a( 10 000 U/ml) had significant anti-proliferative effects and inducing apoptosis on lymphoma cells.Local injection of IFN-a is one of effective therapy on refractory lymphoma.

Adult ; Back Muscles ; physiopathology ; Fasciitis ; physiopathology ; therapy ; Female ; Humans ; Male ; Moxibustion ; Muscular Diseases ; physiopathology ; therapy

Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases. However, due to its complex pathogenesis, there are no officially approved drugs for NAFLD treatment currently. Therefore, it is extremely urgent to find safe and effective anti-NAFLD drugs. Nowadays, lipid-lowering drugs are the main option for NAFLD therapy, but the clinical efficacy of chemical drugs is also very limited, as well as the frequent side effects or adverse reactions. Traditional Chinese medicine (TCM) has attracted more and more attention in the treatment of NAFLD due to its unique advantages through multiple targets and pathways with few side effects. In recent years, numerous studies have demonstrated that the imbalance of gut microbiota plays an important role in the occurrence and development of NAFLD. This review systematically summarizes the experimental and clinical evidences of TCM active compounds and TCM prescription involved in the regulation of intestinal flora in the treatment of NAFLD in recent years, so as to provide a reference for further exploring the pathogenesis of NAFLD and exploring TCM treatment methods.

Fetal atrioventricular block (AVB) is a type of fetal bradyarrhythmias. The reported incidence of fetal complete atrioventricular block (CAVB) and mortality of perinatal fetuses and neonates are signiifcantly higher in pregnancies of anti-SSA/Ro-positive mothers than that of anti-SSA/Ro-negative mothers. The auto-antibodies in maternal serum that can be transported into fetal circulation through placenta may damage fetal cardiac conductive system and eventually result in fetal AVB. There are evidences that early diagnosis and proper treatment can improve the prognosis and survival rate of affected fetuses. In this article, the pathogenesis, risk factors, prenatal diagnosis, treatment and prognosis of fetal immune mediated AVB is reviewed.

Objective To analyze high alumina deformation Micro-CT findings of bone in patients with skeletal fluorosis in Shuicheng Guizhou Province.Methods Bone deformation children,youth and middle-aged patients with fluorosis in Goumi and Zhichang Townships Shuicheng County,coal-burning pollution endemic fluorosis areas,were selected as case group,and non-bone deformation children,youth,and children from non-fluorosis endemic areas as controls.Tibia and the anterior superior iliac spine tissue were obtained through orthopedic surgery and etiology examination,and resin embedded without decalcification.Resin-embedded bone tissue was scanned using micro-CT; relevant parameters were analyzed with ABA special bone analysis software INVEON Research Workplace and three dimensional reconstruction processing software Micview.Results ①The anterior superior iliac spine cancellous bone:compared between bone deformation children and bone non-deformation children in the diseased areas,there was an increasing tendency of the following items:relative volume of trabecular bone(0.337％ vs 0.229％),absolute thickness (μm:139 vs 133),quantities within a unit length (number/mm:2.44 vs 1.72),density woven degree of trabecular bone(number/mm:2.22 vs 1.54) and bone mineral density(mg/cm3:1 033 vs 918),while relative bone area of trabecular bone(mm2/mm3:14.5 vs 15.1) and space pitch (μm:0.274 vs 0.567) declined.Compared between bone deformation youth and bone non-deformation youth in the diseased areas,relative volume of trabecular bone was lower(0.217％ vs 0.437％),relative area increased (mm2/mm3:16.9 vs 11.6),absolute thickness reduced(μm:118 vs 172),trabecular number reduced (number/mm:1.83 vs 2.54),and space pitch increased (μm:0.427 vs 0.222),but density woven degree of trabecular bone increased (number/mm:4.61 vs 1.54),bone mineral density decreased(mg/cm3:977 vs 1 108),osteopenia,osteoporosis,bone mineral decreased,and an increase in the number of trabeculae crossing number.② Tibia bone tissue:compared between bone deformation children and bone non-deformation children in the diseased areas,relative volume of tibia trabecular bone increased(0.435％ vs 0.206％),relative volume of trabecular bone (mm2/mm3:12.3vs 12.4),and thickness (μm:188 vs 161) not changed obviously,trabecular number increased (number/mm:2.43 vs 1.28),space pitch reduced(μm:0.238 vs 0.621),density woven degree of trabecular bone decreased(number/mm:2.40 vs 3.48),bone mineral density increased(mg/cm3:1 047 vs 952),in general presented trabecular thickening,increased number and increased bone mineral.Compared between middle-aged patients with fluorosis in the diseased areas and children in non-fluorosis endemic areas,relative volume of trabecular bone (0.346％ vs 0.206％) and area (mm2/mm3:13.8 vs 12.4) increased,thickness of the trabecular bone reduced (μm:144 vs 161),trabecular number increased (number/mm:1.98 vs 1.28),space pitch decreased (μm:0.318 vs 0.621),and density woven degree of trabecular bone decreased (number/mm:2.60 vs 3.48).Conclusions The results of trabecular bone microstructure and bone mineral density have showed that the combined effects of aluminum and fluorine on human bone tissue at different developmental stages are different.High aluminum and fluorine load before the sexual development of children for trabecular bone thick dense,shows an increasing in bone mass and bone mineral deposition of bone sclerosis image.Bone deformation youth shows osteopenia osteoporosis and bone mineral deposition is reduced.Bone volume is slightly increased,the number of trabecular bone is increasing,trabecular structure is fine in middle-aged patients with skeletal fluorosis.

Objective To explore the effect of electroacupuncture at Hegu (LI 4) on the expressions of P2X2 and P2X3 receptors in experimental pulpalgia rats.Method Forty-two male SD rats were randomized into a normal group (group N), a control group (group C), a pulpalgia model group (group M), an antagonist group (group A), an electroacupuncture group (group E), and an antagonist+electroacupuncture group (group AE), 7 rats in each group. Group N didn’t receive any interventions; group C received injection of normal saline into pulp cavity of the same dose as the injection in group M, and the cavity was then blocked by dental fillings 5-6 min later; in group M, maxillary first and second molar teeth were drilled (drill bit of 1 mm in diameter) to expose pulp and lipopolysaccharide (LPS) solution 5μg/μL was injected into the holes (1~3μL for each hole), and the holes were then covered by dental fillings 5-6 min later; group A received the same modeling method as that in group M, but A-317491 was injected together with LPS (0.5 mg/kg); group E received electroacupuncture at bilateral Hegu (LI 4) with needles retained for 30 min, once a day, totally for 3 times; group AE received the same electroacupuncture intervention after receiving the same treatments as that in group A. The rats’ behaviors and weight were observed for 30 min after intervention each day. The rats were sacrificed on the 4th day, and the mRNA expressions of P2X2 and P2X3 receptors andβ-actin in trigeminal ganglion were detected by using RT-PCR. The mRNA expressions were then compared among the groups.Result The behavioral changes in group M, E, and AE were more significant than that in group C and N (P<0.01); the behavioral changes in group A, E, and AE were less significant than that in group M (P<0.01). The weight in group C was significantly lower than that in group N (P<0.01); the weights in group M, A, E and AE were significantly lower than that in group C and N (P<0.01); the weights in group E and AE were significantly higher than that in group M and A (P<0.01,P<0.05); the weight in group A was slightly higher than that in group M (P<0.05); the weights in group AE was significantly higher than that in group E (P<0.01). The mRNA expressions of P2X2 receptor in group M, A, and AE were significantly higher than that in group N and C (P<0.01); the mRNA expressions of P2X2 receptor in group A, E, and AE were lower than that in group M (P<0.05); the mRNA expression of P2X2 receptor in group A was lower than that in group E (P<0.05); the mRNA expression of P2X2 receptor in group E was higher than that in group AE (P<0.05). The mRNA expressions of P2X3 receptor in group M, A, and E were significantly higher than that in group C (P<0.05) and group N (P<0.01); the mRNA expressions of P2X3 receptor in group A, E, and AE were significantly lower than that in group M (P<0.01); the mRNA expression of P2X3 receptor in group AE was markedly lower than that in group E (P<0.01).Conclusion The expressions of P2X2 and P2X3 receptors in trigeminal ganglion were increased in LPS-induced pulpalgia rats. Electroacupuncture at Hegu (LI 4) and injection of A-317491 both can down-regulate the mRNA expressions of P2X2 and P2X3 receptors, which is plausibly the action mechanism of electroacupuncture at Hegu (LI 4) in analgesia.