Objective:To isolate,culture and identify the human bone marrow mesenchymal stem cells(hBMSC). Methods:Human bone-marrow mononuclear cells(MNC) were isolated by centrifugation over Percoll(1.073 kg/L)and cultured in a BMSC culture system in vitro using their adherent characteristics.Many kinds of cellular molecules were measured to identify the cultured cells.Results:The cells show fibroblast-like irregular appearance,many cytoplasm ecptoma,puff and large nucleus with obvious nucleoli.The flow cytometry shows that 2.09% of the cells are CD34 positive,94.46% of the CD34 negative cells are CD29 positive,and CD54 and CD105 are possibly positive.The cell cycle observation and transmission electron microscopy show that the cells are in the primitive status.The immunocytochemistry detection shows that CD14,CD45 and collagen protein II are negative;CD106,CD166,fibronectin,vimentin and collagen fibers acidic protein(GFAP) are positive;staining for glycogen is strongly positive;staining for alkaline phosphatase is negative.The cells can be induced into osteoblast in proper inductive and differentiating culture circumstances.All the above results indicate that the isolation and culture of hBMSC are successful.Although the hBMSC can be serially subcultivated,the cells appear to be aging and they can not be kept by frozen. Conclusion:The steady ex vivo expansion system in which hBMSC can be obtained has been established.

Objective:To observe clinical therapeutic effect of internal administration of Tongxie Yaofang combined with scalding abdomen with dregs of Shiyi Fang on irritable bowel syndrome(IBS).Methods:75 cases of IBS were treated with internal administration of Tongxie Yaofang combined with scalding abdomen with dregs of Shiyi Fang,and 60 cases treated with western medicine were used as control group.Results:The total effective rate was 80.00% in the treatment group and 51.67% in the control group with a significant difference between the two groups(P

After the research of PLA2R1 and its antibodies, Thrombospondin type-1 domain-containing 7A and its antibodies to membranous nephropathy (MN) has made a new understanding.Some researches have reported that the antibodies of PLA2R1 and THSD7A were mutually exclusive in MN, because THSD7A was found in PLA2R1-negative MN patients.But the latest researcher showed that these antibodies can be both positive in MN patients.Similar to the function of PLA2R1, THSD7A can assist clinical diagnosis, treatment, and monitor of MN.In contrast to PLA2R1, THSD7A was also highly expressed on both human and murine podocytes.We can use the mice model to study the pathogenesis of THSD7A-associated MN in the future.In this review, we describe the structure and function of Thrombospondin type-1 domain-containing 7A and its autoantibodies, highlight its role in MN and suggest possible aspects of its future clinical application.

Objective To explore the curative effects of color Doppter ultrasound guided interventional therapy in pelvic cyst. Methods From July 2006 to June 2008,the color Doppler ultrasound guided transabdominal or transvaginal puncture was performed in 34 pelvic cysts among 32 diagnosed cases,absolute alcohol was injected into the cysts for sclerotherapy.Results Among 34 cysts,the treatment only failed in one cyst of one case because of the over ropy hydatid fluid,which was hard to be aspirated.All of the cases were followed up for three months.Twenty-seven cysts were cured after the first treatment,the cure rate was 79.4%(27/34).The maximum diameter of other 4 cysts decreased more than 1/2.The effective rate was 91.2%(31/34) after the first puncture.Finally,33 cases showed curative effect after the puncture therapy,the total effective rate was 97.1%(33/34).No serious complications were observed.Conclusion This therapy is a simple,accurately,safe and effective treatment for many indications.It's worthy to be recommended in the therapy of pelvic cyst.

With the development of peaple's life and the change of diet,the incidence rate of colorectal cancer is increasing. There are Twenty-five percent of patients were found liver metastases in the first diago-sis. Surgical resection of liver metastases from colorectal cancer is known to be associated with long term sur-vival. So it is the key to increase the resection rate for colorectal cancer patients with liver metastases. The in-curruence of neoadjuvant therapy may be useful in therapy of colorectal cancer patients with liver mtastases.

Objective To evaluate the efficacy of dynamic moxibustion plus acupuncture in treating cervical vertigo by clinical observation of vertebrobasilar artery blood flow velocity using transcranial color Doppler before and after treatment. Method Sixty-one patients diagnosed with cervical vertigo were randomly allocated to two groups. The treatment group of 31 patients received dynamic moxibustion plus acupuncture and the control group of 30 patients, acupuncture alone. Vertebrobasilar artery blood flow velocity was observed using transcranial color Doppler before and after treatment. Result The total efficacy rate was 96.8% in the treatment group and 73.3% in the control group. Vertebrobasilar artery blood flow velocity was improved in both groups, but the effect was more marked in the treatment group (P<0.05). Conclusion Dynamic moxibustion plus acupuncture can more effectively improve vertebrobasilar artery blood flow velocity than acupuncture alone.

Ca2+signaling is fundamental for information process-ing in the peripheral nervous system,which regulates a variety of physiological activities.Ca2+signaling and calcium homeostasis are directly associated with neuropathology.Recently,studies on Ca2+signaling contribute to a deeper comprehension of the path-ogenesis of diabetic peripheral neuropathies,which provide a new research direction for the treatment of diabetic peripheralneuropathies.This review aims to highlight the relationship be-tween calcium signaling,sensory neurones and neuroglial cells in the context of diabetic peripheral neuropathies.

ObjectiveTo observe the treatment possibility and effects on rats models with experimental femoral artery obliterans(EFAO) by transplantation of endothelial progenitor cells(EPC) from bone marrow. MethodsEFAO rat models were successfully made. Bone marrow mesenchymal stem cells(MSCs) were isolated from allogeneic male Wistar rat,cultured in DMEM in vitro. EGF,bFGF and IGF-1 were added into culture medium on the 10th day and the active EPC were labled with BrdU. 5×106 of the above cells were transplanted into the right hindlimbs i.m. as Group A in the 16 rats,and the same volume of normal saline into the opposite as Group B as control. Laser Doppler perfusion imaging was taken on the 0 day、the 14th day and the 28th day after transplantation. On the 14 day and the 28 day 8 rats were sacrificed respectively and muscles of all hindlimbs were extracted for immunohistochemical examinations (by FⅧ and BrdU).Results1. In group A the skin blood perfusion in hindlimb were significantly increased as compared with group B(P<0.05). 2. Some positive stained EPC by BrdU were found in Group A but not in Group B. 3. The numbers of blood vessels with positive staining of FⅧ in hindlimb on the 28 day in Group A were more obviously than that in Group B (P<0.01). ConclusionTransplantation of EPC from bone marrow can significantly increase the skin blood perfusion and capillary density in ischemic hindlimbs in EFAOrats.

Background Inflammation and adipogenesis are two parallel processes with increasing activity in severe thyroid-associated ophthalmopathy(TAO),and mevastatin was proved to have the inhibiting effect on the differentiation of adipose.Objective The aim of this work was to investigate the effects of mevastatin on the expression of cyclooxygenase-2(COX-2)and peroxisome proliferator activated receptor-γ(PPAR-γ)and differentiation of TAO-derived orbital preadipocytes,and explore its modulation effects on lipopolysaccharide(LPS)induced inflammation and the differentiation of TAO-derived orbital preadipocytes in vitro.Methods The retroorbital adipose tissue was obtained from 4 TAO patients during the surgery.The orbital fibroblasts were cultured from orbital adipose tissues using explant culture method.To study the suppressing effect of mevastatin on inflammatory response,cultured cells were divided into 5 groups.The 1000 μg/L LPS orbital fibroblasts were stimulated for 8 hours in group A,and 1000 μg/L LPS combined with 5 μmol/L,10 μmoL/L or 20 μmoL/L mevastatin were used respectively for the substitute in the group B,group C and group D.The orbital fibroblasts in group E were cultured routinely without any intervention as control.To observe the inhibiting effect of mevastatin on the differentiation of adipose,the group A were then subdivided into group A1-A6.After 1000 μg/L LPS was used to treat the cells for 8 hours,the ceils were induced to differentiate into adipocytes.All orbital preadipocytes from A1 to A6 were stimulated to differentiate into mature adipocytes with cocktail differentiation medium for a 10-day duration.During the procedure,group A2,A3 and A4 were interfered with 5,10 or 20 μmol/L mevastatin,and in the group A5 and A6,10 μmol/L mevastatin were added at the fourth day or eighth day.Intracellular fat accumulation in differentiated adipocytes was determined by oil red O staining.The absorption(A492 nm)was measured in the ceils by enzyme-linked immunosorbent assay(ELISA).Expression of COX-2 and PPAR-γ mRNA was detected by reverse transcription polymerase chain reaction(RT-PCR),and the expression of COX-2 and PPAR-γ protein was detected by Westernblot.The level of PGE2 in the supernatant was detected by ELISA.Results The expression of COX-2 protein and mRNA as well as the PGE2 levels in B,C,D group decreased markedly in comparison with those in A group(P＜0.05).With the increase of mevastatin concentration,the expression of COX-2 protein and mRNA as well as the PGE2 levels in B,C,D groups decreased successively(F =228.380,101.745,1586.881,P＜0.05).The expression of COX-2 protein and mRNA and PGE2 levels in E group were lower significantly than those in A,B and C groups(P＜0.05),but no significant differences were found between E group and D group(P＞0.05).The A492 value and the expressions of PPAR-γ protein and mRNA in differentiated cells showed the successively decrease in A1-A4 group with the elevation of mevastatin concentration(P＜0.05),and the evidently decreased A492 value and the expressions of PPAR-γ protein and mRNA also were seen in A1 and A5 groups compared with A3 group(P ＜ 0.05).Conclusions Mevastatin inhibits LPS-induced COX-2 expression,PPAR-γ expression,PGE2 secretion and differentiation of TAO-derived orbital fibroblasts in vitro in dose-dependent manner.Mevastatin plays these effect more prominently in early stage of adipocytes differentiation.

Background The orbital fibroblasts (OFs) in thyroid associated ophthalmopathy (TAO) play important roles in the proliferative and inflammatory response.Seeking the drug which inhibit OFs growth is of a vital significance for the prevention and treatment of TAO.Research documented that colchicine has an anti-fibrosis effect.But its influence on OFs of TAO patient is few known.Objective This study was to investigate the effect of colchicine on growth and apoptosis of OFs in vitro.Methods The retroobital connective tissue was obtained form 3 TAO patients and cultured using explant method.OFs were passaged and identified by immunochemistry,and 3-8 genetaions of cells were used in the study.Colchicine at the concentrations of 1 × 10-8,1 × 10-7,1 × 10-6,1 × 10-5,1 ×10-4 mol/L was added into the RPMI 1640 with 10％ fetal bovine serum(FBS) to incubated the cells for 24,48 and 72 hours respectively,and only RPMI 1640 was used to culture the cells as the control group.Cell counting kit-8 (CCK-8)was used to detect the absorbance value (A450) of OFs for the evaluatuion of OFs and the inhibitory rate of colchicine to OFs.The colchicine of 1 ×10-6,1 ×10-5,1 × 10-4 mol/L was added into the culture medium for 48 hours,and then the apoptotic rate of the cells and the cell percentage in various cellular cycle was assayed by flow cytometry(FCM).The expression of transforming growth factor-β (TGF-β)in the cells was detected by immunochemistry to assess the influence of colchicine on the serection of the cells.Results Cultured cells showed the spindle-like in shape and the cell number was significantly increased with the incubation time.After incubated with 1 × 10-4,1 × 10 5,1 × 10-6,1 ×10-7,1 × 10-8 mol/L colchicines,the A450 values were gradually reduced with the increase of the concentrations of colchicine(F ion =62.004,P＜0.05),and significant differences were found between different contrations of colchicine groups(all P＜0.05).Aslo,gradually declined A450 values of the cells were seen with the lapse of culture time among the groups(Ftime =459.582,P＜0.05).The inbitory rate of colchicine to the cells was elevated with the increase of concentrations.The apoptotic rates of the cells were (1.73 ± 0.15) ％,(21.04 ± 4.56) ％,(31.84 ±6.21)％and(35.32±5.56)％ in the control group and 1 × 10-6,1 × 10-5,1 × 10 4 mol/L colchicine groups respectively,with statistically significant difference among the 4 groups (F =83.905,P＜0.05).With the increase of concentrations of colchicines,the cell percentage in G2 +M phase lessened gradually,showing significant difference among the control group and the 1 × 10-6,1 × 10-5,1 × 10-4 mol/L colchicine groups (F =20.443,P＜0.05).The expression of the TGF-β in the cells was (97.60± 2.09) ％ in the control group,and that in the 1 × 10-4 mol/L colchicine group was (44.43 ± 3.96) ％,presenting a significant difference between them (t =65.330,P ＜ 0.05).Conclusions Colchicine can induce apoptosis of OFs and inhibit the prolilferation of OFs in a time-and dose-dependent manner probably by decreasing the TGF-β secretion