OBJECTIVE:To provide approaches and avenues for the Chinese drugs preparations manufacturers in the GMP depuratory authentication.METHODS:The features and difficulties in the GMP depuratory authentication of the Chinese drugs preparations such as the optimizing of depuratory regulations,the determination of the acceptable standards,the choice of testing methods,clarification of expiration date and re-authentication were analyzed.RESULT&CONCLUSION:The depuratory regulations of the manufacturing facilities for the Chinese drugs preparations must be optimized.

OBJECTIVE:To provide references for the improvement of our country's current Good Manufacturing Practice(GMP).METHODS:In consideration of GMP standards and problems in its implementation,the system,customers,suppliers,business leaders and documentation etc.were analyzed.RESULTS&CONCLUSION:Problems in the GMP standards has influenced the practice of GMP.In order to improve and optimize the GMP standards,it was recommended that process system be integrated to make the design quality in conformity with the manufacturing quality,open quality management system be established,business leader's leading effect be emphasized and clear and universal documentation system be established.

Nanomaterials appear to be promising for a number of applications in in-vitro diagnostics, mainly due to the biocompatibility, specific surface and conduction activity. The use of nanostructures as diagnostic tools has the advantage of very low limit of detection achievable, high sensitivity, strong selectivity and the possibility to fabricate point-of-care diagnostic devices. With the problems of nanotechnology solved, nanomaterials had already put a broad impact in the field of clinical medical examination.
Nanostructures ; Nanotechnology

BACKGROUND:Diabetic foot ulcers threaten the patients’ health and even survival seriously. It is an international difficult problem and lacks an effective treatment. But gene therapy and stem celltherapy possess special advantages and potential in wound healing. OBJECTIVE:To assess the therapeutic effect of transplantation of bone marrow mesenchymal stem cells transfected by human vascular endothelial growth factor 165 (hVEGF165) gene on foot wound healing in diabetic rats. METHODS:Recombinant adenovirus was established in vitro which expressed hVEGF165 gene and transfected into the third generation of bone marrow mesenchymal stem cells. Total y 120 male Wistar rats were divided into five groups:group A (non-diabetic controls), group B (diabetic controls), group C (Ad-hVEGF165 therapy), group D (stem celltherapy) and group E (transplantation of bone marrow mesenchymal stem cells transfected by Ad-hVEGF165 gene). Rats in the latter four groups were intraperitoneal y injected with streptozotocin to induce diabetic models. In al rats, a 3 mm×7 mm rectangular ful-thickness skin sample was cut from the instep of the hind foot to make a model of foot wound. The rats were subcutaneously injected at equidistant six points 5 mm distal to the wound edge on the dorsum of the foot:50μL PBS per point for group A, 50μL adenovirus suspension (1×1013 pfu/L) per point for group C, 50μL stem cellsuspension (1×1010/L) per point for group D, and 50μL adenovirus suspension+50μL stem cellsuspension per point for group E. RESULTS AND CONCLUSION:After injection, the rate of wound healing, the expression of VEGF and the qualities of capil aries in group E were higher when compared with groups B, C, D (P<0.05), but were lower than those in group A (P<0.05). Transplantation of bone marrow mesenchymal stem cells transfected by hVEGF165 gene can promote foot wound healing, angiogenesis and expression of VEGF in diabetic rats.

Objective To investigate the clinical efficacy for different treatments of stage Ⅳ right colorectal cancer and its prognostic factors.Methods The clinical data of 106 patients with stage Ⅳ right colorectal cancer who were admitted to the First People's Hospital of Chenzhou from January 2008 to December 2013 were retrospectively analyzed.Among the 106 patients,42 patients receiving palliative resection were allocated to the palliative resection group,30 patients receiving colostomy were allocated to the colostomy group,20 patients receiving bypass surgery were allocated to the bypass group and 14 patients without treatment were allocated to the non-treatment group.Fluorouracil + leucovorin (5-FU/LV) were used as postoperative chemotherapeutics,and the time of chemotherapy was 2 to 6 months.The follow-up was applied to the patients by outpatient examination and telephone interview till February 2014.The non-normal distribution data were described as median and range.The survival curve was drawn by Kaplan-Meier method,and the survival rate was analyzed using the Log-rank test.The continuous variables were cut into the categorical variables.The univariate analysis of categorical variables was done using chi-square test,and the multivariate analysis was done using the COX regression model.Results The incidences of postoperative complications in the palliative resection group,in the colostomy group and in the bypass group were 47.6％ (20/42),40.0％ (12/30) and 65.0％ (13/20),respectively,with no significant difference (x2=3.053,P ＞ 0.05).One-hundred patients were followed up for 14.0 months (range,3.0-40.0 months),with overall median survival time of 10.3 months (range,2.6-27.0 months) and the 1-,2-year survival rates of 36.8％ and 6.7％.The median survival time and 1-,2-year survival rates were 11.5 months (range,4.3-27.0 months),47.6％,16.7％ in the palliative resection group,8.5 months (range,3.5-18.0 months),20.0％,0 in the colostomy group,9.0 months (range,3.0-13.0 months),15.0％,0 in the bypass surgery and 5.0 months (range,2.6-10.0 months),0,0 in the non-treatment group,showing a significant difference in the prognosis of patients among the 4 groups (x2 =42.395,P ＜ 0.05).The prognosis of patients in the palliative resection group were significantly different from those in the other 3 groups (x2 =5.786,6.178,10.378,P ＜0.05),there was no significant difference in the prognosis of patients between the colostomy group and the bypass surgery group (x2 =0.203,P ＞ 0.05).The results of univariate analysis showed that T stage,N stage,tumor differentiation,preoperative obstruction,peritoneal implantation,methods of treatment,chemotherapy and postoperative complications were related factors affecting the prognosis of patients with stage Ⅳ right colorectal cancer (x2=37.428,48.586,32.550,22.739,33.562,42.395,21.517,11.530,P＜0.05).T4 stage,N2 stage,poor-differentiated tumors and peritoneal implantation were independent risk factors affecting the poor prognosis of patients with stage Ⅳ right colorectal cancer (RR =2.336,2.945,2.182,3.500,95％ confidence interval:1.102-4.953,1.156-7.501,1.003-4.749,1.573-7.787,P ＜0.05).The postoperative chemotherapy was an independent factor affecting the good prognosis of patients with stage Ⅳ right colorectal cancer (RR =0.495,95％ confidence interval:0.271-0.904,P ＜ 0.05).Conclusion Palliative resection can improve the prognosis of patients with stage Ⅳ right colorectal cancer.T4 stage,N2 stage,poor-differentiated tumor,and peritoneal implantation were independent risk factors affecting the poor prognosis of patients with stage Ⅳ right colorectal cancer,while postoperative chemotherapy was an independent factor affecting the good prognosis of patients with stage Ⅳ right colorectal cancer.

OBJECTIVE: To assess the therapeutic effect of treating diabetic foot ulcers (DFUs) in rats by subcutaneously transplanting around the wounds and intramuscularly into the leg with bone marrow derived mesenchymal stem cells (BM-MSCs). METHODS: BM-MSCs from male Wistar rats were cultured by the whole bone marrow adherence method until the third generation. The BM-MSCs were labeled by 4,6-diamino-2-phenylindole (DAPI) in vitro. Forty-eight male Wistar rats were randomly divided into 4 groups: group A (n=12), rats with DFUs receiving BM-MSCs subcutaneous transplantation; group B (n=12), rats with DFUs receiving BM-MSCs intramuscular transplantation; group C (n=12), nondiabetic rats with foot ulcers; and group D (n=12), rats with DFUs receiving no BM-MSCs. A diabetic rat model was induced by intraperitoneally injecting streptozotocin (STZ). Then DFU model was established by removing a 3 mm × 7 mm rectangular full-thickness skin on the 2 back dorsum pedis surfaces. On day 2, 5, 8 and 11 after the transplantation, the rate of wound closure was raised; trace of DAPI labeled-BM-MSCs in the wound tissues was observed on frozen sections and the thickness of granulation tissues was detected by HE stain. Immunohistochemistry was performed to detect the expression of CD31 and Ki-67. The expression of vascular endothelial growth factor (VEGF) in the wound tissues was detected by ELISA and RT-PCR. RESULTS: The wound closure in group C was faster than in other groups (P<0.05). The rate of wound healing in group B was higher than group A on day 11 (P<0.05). The intensity and area of the fluorescence in group B were higher than group A on day 2 and 5 on the frozen sections. HE stain showed that the granulation tissue formation in group B was thicker than group A on day 5. Immunohistochemistry of CD31 demonstrated that the mean number of small blood vessels in group B was more than in group A on day 5 and 8 (P<0.05). Immunohistochemistry of Ki-67 showed it had no difference between group A and group B. ELISA and RT-PCR revealed that the expression level of VEGF in the wound tissues in group B was higher than in group A on day 8 and 11 (P<0.05, P<0.001, respectively). CONCLUSION Both transplantations promote the wound healing of DFUs in rats. The intramuscular transplantation into the leg shows a better persistence and a higher expression level of VEGF in the wound tissues at later stages.
Animals ; Bone Marrow Cells ; cytology ; Diabetes Mellitus, Experimental ; complications ; Diabetic Foot ; therapy ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Rats ; Rats, Wistar ; Vascular Endothelial Growth Factor A ; metabolism ; Wound Healing

OBJECTIVETo assess the therapeutic effect of intramuscular transplantation of bone-marrow derived mesenchymal stem cells (BM-MSCs) in the leg for treatment of diabetic foot ulcers (DFUs) in rats.

METHODSThirty-six male Wistar rats were randomly allocated into 3 equal groups, namely group A with DFUs on the bilateral hindlimb dorsum pedis and intramuscular transplantation of 4,6-diamino-2-phenylindole (DAPI)-labeled third-passage BM-MSCs from male Wistar rats into the leg, group B with nondiabetic foot ulcers, and group C with DFUs but without BM-MSC transplantation. On days 2, 5, 8 and 11 posttransplantation, the rate of wound healing was evaluated, the labeled BM-MSCs in the wound tissues were traced on frozen sections, and the thickness of granulation tissues was measured with HE staining. Immunohistochemistry was performed to detect the expression of CD31 and Ki-67, and the expression of vascular endothelial growth factor (VEGF) in the wound tissues was detected by ELISA and RT-PCR.

RESULTSThe rats in group B showed significantly faster wound closure than those in the other two groups (P<0.05). The rate of wound healing was greater in group A than in group C on days 8 and 11 (P<0.05). The fluorescence intensity and area on the frozen sections were the highest on day 5 posttransplantation in group A, which had a granulation tissue thickness comparable with group B but greater than group C. The mean numbers of small blood vessels and cells positive for CD31 and Ki-67 expressions were similar between groups A and B, which showed significant differences from group C (P<0.05). On day 11, group A showed the highest VEGF expression in the wound tissues (P<0.001).

CONCLUSIONIntramuscular transplantation of BM-MSCs can significantly promote wound healing of DFUs in rats possibly as a result of increased expression of VEGF in the wound tissues.

Animals ; Bone Marrow Cells ; Diabetic Foot ; surgery ; Lower Extremity ; Male ; Mesenchymal Stem Cell Transplantation ; Muscle, Skeletal ; transplantation ; Rats ; Rats, Wistar ; Treatment Outcome ; Wound Healing

Nanomaterials appear to be promising for a number of applications in in-vitro diagnostics, mainly due to the biocompatibility,specific surface and conduction activity. The use of nanostructures as diagnostic tools has the advantage of very low limit of detection achievable, high sensitivity,strong selectivity and the possibility to fabricate point-of-care diagnostic devices. With the problems of nanotechnology solved,nanomaterials had already put a broad impact in the field of clinical medical examination.

AIM: To investigate the role and possible mechanism of gastrodin combined with dexamethasone in myocardial cell injury induced by oxygen-glucose deprivation. METHODS: Oxygen-glucose deprivation (OGD) model was established. The cells were divided into 5 groups: normal control group, OGD group, DEX group, GAS group and DEX+GAS group. The activity of myocardial cells was detected by CCK-8 test in each group. The activity of LDH was detected by colorimetry in each group. The apoptosis of myocardial cells was detected by TUNEL method in each group. The ELISA assay was used to detect the inflammatory factors in culture medium of myocardial cells in each group. Western blot was used to detect the expression of Notch1, Bax, Bcl-2 and Beclin1 in myocardial cells in each group.RESULTS: The results showed that GAS combined with DEX could significantly increase the activity of myocardial cells and decrease the apoptosis, reduce production of TNF-α, IL-6, IL-1β and promote production of IL-10, decrease the release of LDH significantly of myocardial cells induced by OGD. The results of Western blot showed that GAS combined with DEX increased the expression of Notch1, Bcl-2 and autophagy-related gene Beclin1, but decreased the expression of Bax of myocardial cells induced by OGD. CONCLUSION: The combination of GAS and DEX may promote autophagy and increase cell activity, inhibit apoptosis and inflammatory reaction by activating Notch signaling pathway, thereby reducing OGD-induced myocardial cells damage.