Objective To optimize prescription for double-layered Erhuang sustained-release suppository. Methods Amounts of PEG400, PEG4000, HPMC were selected as influence factors for L9(34) orthogonal experiment. A comprehensive assessment was conducted by setting the cumulative release degree at three different time points as index, and the inner and outer layers of double-layered Erhuang sustained-release suppository were optimized. Results The best prescription was the inner HPMC∶PEG4000∶PEG400=1.5∶10∶4;outer HPMC∶PEG4000∶PEG400=0.5∶10∶4. Conclusion Prescription for double-layered Erhuang sustained-release suppository has good forming property and a good sustained-release effect according to the optimized prescription, which has certain reference value for researches and development of TCM suppository.

In order to breed and spread a new cultivar of Curcuma wenyujin, the C. wenyujin germplasm resources were investigated in authentic regions. Better varieties were chosen by comparing the yield, economic characters and quality differences between different cultivars. The results showed that the character of new selected cultivar was stable, the yield of zedoary, turmeric and curcuma was reached 313.7, 177.9, 91.2 kg per 667 m2, respectively, it increased 11.6%, 10.2%, 14.2% comparing with farmer varieties. The volatile oil contents in zedoary and turmeric was 4.0%, 3.0%, respectively. The target ingredients (germacrone) content was stable. It is demonstrated that the new cultivar "Wenyujin No. 1" has value for extension at authentic regions.
Breeding ; China ; Curcuma ; chemistry ; growth & development ; Oils, Volatile ; analysis ; Plant Extracts ; analysis

Emerging evidence has indicated that circular RNAs (circRNAs) play pivotal roles in the regulation of cellular processes and are found to be aberrantly expressed in a variety of tumors.However,the clinical role of circRNAs in bladder cancer (BC) and the molecular mechanisms have yet to be fully understood.In this study,the clinical specimens were obtained and the expression level of a circRNA BCRC4 was detected by real-time PCR in both BC tissues and cell line.The circular RNA over-expression plasmid was constructed and transfected into BC cells and related cell line.The cell cycles and apoptosis were observed using inverted microscope and flow cytometry.Western blotting was used to compare the relative protein expression of groups with different treatments.It was found that circRNA BCRC4 expression was lower in BC tissues than in adjacent normal tissues.Furthermore,consequences of fomed-expression of BCRC4 promoted apoptosis and inhibited viability of T24T and UMUC3 cells,and up-regulated BCRC4-inereased miR-101 level,which suppressed EZH2 expression in both RNA and protein levels.In addition,gambogic acid (GA) is a promising natural anticancer compound for BC therapy,and GA treatment increased the BCRC4 expression in T24T and UMUC3 cells in a dose-dependent manner.Altogether,our findings suggest that BCRC4 functions as a tumor suppressor in BC,and mediates anticancer function,at least in part,by up-regulating the expression of miR-101.Targeting this newly identified circRNA may help us develop a novel strategy for treating human BC.

Objective To explore the possibility of basic fibroblast growth factor(bFGF)and epidermal growth factor(EGF)combined with striatal conditioned medium promoting the directional differentiation of rat bone marrow mesenchymal stem cells(BMMSCs)into dopaminergie neurons.Methods 1.Separation and culture of BMMSCs:BMMSCs were harvested from healthy adult Wistar rats for serial subcultivation.2.Preparation of Striatal conditioned medium:newborn Wistar rats within 24 hours were selected,and their brain tissues were removed to prepare striatal conditioned medium.3.Induced differentiation of BMMSCs:the 5th passage BMMSCs were collected and pre-induced in low glucose-Dulbecco's modified eagle medium(L-DMEM)containing bFGF and EGF.Twenty-four hours later,pre-induction liquor was replaced with striatal conditioned medium for further induced differentiation.4.Result assessment:the morphological changes of stem cells were observed under inverted phase microscope.The expression of neuron specific enolage(NSE)and tyrosine hydmxylage(TH)were identified by immunocytochemical technique.Results The cell body of rat BMMSCs contracted into round and spindle shape after induction by bFGF and EGF combined with striatal conditioned medium.Partial neuron-like cells with prominence could be found.Immunocytochemieal detection showed that the percentages of NSE and TH positive cells were(72.70±14.81)% and(34.50±15.93)%,respectively.Conclusion BMMSCs can be induced directionally into dopaminergiC neurons by bFGF and EGF combined with striatal conditioned medium in vitro.

Objective　To investigate the molecular regulation of G1 arrest of mouse thymocytes induced by ionizing radiation. Methods　Cell cycle was analyzed by flow cytometry (FCM) following staining of cells with proidium iodide. Fluorescent staining and flow cytometry analysis were employed for measurement of protein expression. Results　It was demonstrated that G1 phase of mouse thymocytes increased significantly at 12h after whole body irradiation (WBI) with the doses of 0.5, 1.0 and 2.0 Gy, and at 24h following 2.0Gy exposure, measured by FCM. In the time course experiment, it was found that G1 phase of thymocytes increased significantly at 4h, reached a peak level at 24h and came down toward 48h after WBI with 2.0Gy X-rays. The results also showed that after 2.0Gy exposure, the expression of proteins in mouse thymocytes increased significantlty from 1h to 8h for p53, for p21 from 4h to 48h, and for MDM2 at 4h and 8h, measured by FCM. But no change was found for GADD45 protein expression. Conclusion　These results suggest that G1 arrest could be induced by a single dose of 0.5 Gy, 1.0Gy or 2.0Gy, and its molecular control might be established through the p53-p21 pathway.

AIM: To investigate the immunologic mechanism of CXC chemokine ligand 10(CXCL10) and its receptor CXC chemokine receptor 3 (CXCR3 ) involved in the process of endometriosis (EM). METHODS: Serum samples were collected from 3 groups; EM patients without operation (n = 76) , EM patients with operation (n = 10) and the normal control persons (n =76). CXCL10 and CA12S concentrations were detected by means of ELISA and chemilumino-metry. Cell surface antigens on the activated PBMC - CD3 and CXCR3, as well as CXCR3 subgene - CXCR3A and CX-CR3B were tested by flow cytometry (FC) and RT - PCR when PBMC was separated from women with EM ( n = 10) and without EM (n = 10), and then activated. RESULTS: Serum CXCL10 concentrations between three groups were signifi-canly different (P < 0.05). Compared to normal control group, although the supernatant CXCL10 concentration and CD3~+ /CXCR3~+ PBMC number in EM group has no significant difference (P >0.05) , highly expressed CXCR3B in EM group rather than CXCR3A was observed. CONCLUSION: CXCL10 in women with EM is low, indicating that it plays a vital role in the process of EM and immune system of the women with EM is defected and impaired. The immunoreactivity of PBMC from both EM patients and normal person is same to activated signal, but the productions are different: PBMC in EM group mainly express CXCR3B but PBMC in normal person mainly express CXCR3A after activation, which may be one of the immune mechanisms that EM escapes from immunological lethal effect of the infected host.

OBJECTIVETo study the effect of gene radiotherapy combining injection of recombinant plasmid pNEgr-mIL-12 with local X-irradiation on cancer growth and to elucidate the mechanisms of tumor inhibition.

METHODSAlkaline lysis was used to extract the plasmid and polyethylene glycol 8000 (PEG 8000) was applied for further purification of plasmids. Enzyme-linked immunosorbent assay (ELISA) was used to detect the expression of IL-12 protein. C57BL/6J mice were subcutaneously inoculated with B16 melanoma cells and the plasmid was injected directly into the tumor. Gene-radiotherapy combining pNEgr-mIL-12 recombinant plasmid with X-irradiation was given three times to C57BL/6J mice bearing B16 melanoma. Changes in immunologic parameters of tumor-bearing mice were detected with relevant immunologic assays.

RESULTSResults showed a significant decrease in tumor growth rate (P<0.05-0.001) after 3 times of gene-radiotherapy with IL-12 and X-irradiation. Immunologic studies showed a significant increase in CTL and NK cytolytic activity (P<0.05-0.001) and an up-regulated secretion of IFN-gamma and TNF-alpha (P<0.01-0.001). Moreover, the expression of mIL-12 in B16 melanoma cells of the treated tumor-bearing mice was found to be higher than that of control.

CONCLUSIONpNEgr-mIL-12 plasmid combined with X-irradiation can increase tumor control and the mechanism of increased tumor inhibition is related to the enhancement of anticancer immunity in tumor-bearing mice.

Animals ; Combined Modality Therapy ; Enzyme-Linked Immunosorbent Assay ; Female ; Genetic Therapy ; Interferon-gamma ; immunology ; Interleukin-12 ; biosynthesis ; genetics ; therapeutic use ; Killer Cells, Natural ; immunology ; Macrophages ; immunology ; Melanoma, Experimental ; immunology ; radiotherapy ; therapy ; Mice ; Mice, Inbred C57BL ; Plasmids ; Spleen ; immunology ; T-Lymphocytes, Cytotoxic ; immunology ; Tumor Necrosis Factor-alpha ; immunology ; X-Rays

Objective To provide vascular feature of neurocutaneous vascular flap and anatomical details about how to design the flap.Methods Ten fresh human body sample with twenty limb were perfused.The clinical anatomy of lateral antebrachial cutaneous nerve,medial antebrachial cutaneous nerve,sural nerve,superficial peroneal nerve,saphenous nerve and their nutrient vessels were studied.The distribution of their nutrient perforators were observed.Results Neurocutaneous nutrient vessels or nutrition artery with large diameter were accompanying nervus cutaneus by a long distance; Or longitudinal vascular chains were formed by ramus communicans with the ascending branches and descending branches from multiple segmental vessels.Medial antebrachial cutaneous nerve,lateral antebrachial cutaneous nerve,sural nerve,superficial peroneal nerve,saphenous nerve has the ulnar artery perforating branches,radial artery perforating branches,anteriolateral supra malleolar perforating branches,posterolateral supra malleolar perforating branches,medial supra malleolar perforating branches,accompanied separately,and the occurrence rate were 100％,95％,80％,90％,100％ respectively.Conclusion Cutaneous branch from the main deep artery is the anatomical basis of neurocutaneous nutrient vessel.Its distribution also accord to pressure balance rule.Mostly nervus cutaneus had constant perforator attending to suply its nutrition.Actually neurocutaneous nutrient vessel is a predictable and reliable vascular chain.

Objective To explore the use of CA199,CEA,CA50 and ALP combined with magnetic resonance imaging (MRI) in diagnosis of intrahepatic bile duct stones with early bile duct carcinoma.Methods The clinical diagnosis of 36 cases of patients with hepatolithasis-associated intrahepatic cholangiocarcinoma (HICC) and 118 cases with intrahepatic duct stone (IHDS) were analyzed retrospectively.Serum CA199,CEA,CA50,ALP and magnetic resonance (MRI,MRCP) were performed and the results were analyzed.Results Abdominal pain discomfort in Hepatolithasis-associated intrahepatic cholangiocarcinoma showed multiple symptoms.Three typical Charcot syndromes were rare.On the gender distribution,intrahepatic bile duct stones was frequently found in women,on the contrary,Hepatolithasis-associated intrahepatic cholangiocarcinoma was frequently more found in men (P ＜0.05) Abdominal pain and fever in patients of Hepatolithasis-associated intrahepatic cholangiocarcinoma was more than that of patients with Intrahepatic bile duct stone (P＜0.05) When alkaline phosphatase (ALP) was more than 169 U/L,significant difference was seen between two groups (P＜ 0.05) According to the diagnosis of HICC,the accuracy of CA199,CEA,CA50 combined with ALP was 88.6％,the accuracy of magnetic resonance examination alone was 90.2％,and the accuracy of multiple serological markers and magnetic resonance was 95.5％.Conclusion MRI combined with serum CA199,CEA,CA50 and ALP can effectively improve the HICC preoperative diagnosis rate.

Objective To investigate the potential application of targeting at vascular endothelial growh factor (VEGF) induced apoptosis in leukemic cell lines by combined use of Bevacizumab and chemotherapeutic drug. Methods Leukemic cells were treated with several drugs at different concentrations in culture. The effect of VEGF, Bevacizumab and co-treated with Ara-C on leukemic cells proliferation were evaluated by CCK-8 and apoptosis and cell cycle were detected by flow cytometry (FCM). Results VEGF could enhance the proliferation of leukemic cells and caused a dose-dependent manner on U937 cell. It also increased the percentage of cells in S phase, tested by, and Bevacizumab group was decreased. Apoptotic rate of cells treated with Bevacizumab or co-treated with Bevacizumab and Ara-C for 48 h were significantly higher when compared with control or Ara-C group, respectively (P<0.05), but the apoptotic rate of VEGF group or VEGF and Ara-C group was lower (P>0.05). There was no significant difference in apoptotic rate between control and combined use of VEGF, Bevacizumab and Ara-C group(P>0.05). Conclusion VEGF could enhance the proliferation of some leukemic cells, and may contribute to leukemic cells survival and a resultant resistance to chemotherapy-triggered cell death. The study also showed that leukemic cells growth was significantly inhibited by Bevacizumab through directly against VEGF, and the sensitivity of leukemic cells for chemotherapeutic drug was increased.