OBJECTIVE: To establish a highly effective HPLC method for analyzing 1-chlorine-2,4-dinitrobenzene (CDNB), and to determine the activity of GST enzyme in rat liver microsomes (RLM) by use of CDNB as probe and study its enzyme kinetics. METHODS: Welch Materials Ultimate™ XB C18 column (4.6 mm × 250 mm, 5 μm) was used, the mobile phase was acetonitrile-water(7:3), the flow rate was 0.8 mL · min-1, and the detection wavelength was 238 nm. CDNB was incubated with 0.02 mg · mL-1 protein in RLM at 37°C for 7 min and the reaction was terminated by cool acetonitrile. The reaction solution was centrifugated and the supernatant was filtered and analyzed by HPLC. The Vmax, Km and CLint were calculated by Sigma Plot. RESULTS: The retention time of CDNB was about 6 min without any interference. The lowest detection limit of CDNB was 1.0 μmol · L-1, and the calibration curve was linear from 2.5 to 100.0 μmol · L-1. The intra-day and inter-day relative standard deviations were less than 10% respectively. The method recoveries ranged from 99.38% to 108%. The reaction catalyzed by GST was terminated by acetonitrile and the incubation time was 7 min. The kinetic parameters, Vmax, Km, and CLint were 85.45 nmol · min-1 · (mg protein)-1, 15.09 μmol · L-1, and 5.73 mL · min-1 · (mg protein)-1, respectively. CONCLUSION: This method is reliable and the results can ac-curatelly indicate GST enzyme activity, which can be used for the kinetics study of GST in RLM.

·AIM:To investigate the effect of β1-integrin overexpression on the adhesion and migration of rabbit corneal epithelial (RCE) cells.·METHODS:Eukaryotic expression vector encoding β1-integrin-GFP fusion DNA was transfected into RCE cells,and the β1-integrin-GFP fusion gene was examined by RT-PCR and Western blot.The adhesion to Matrigel and the migration of the transfected cells were determined by adhesion and mobility assays.The phosphorylation of focal adhesion kinase (FAK) was examined by Western blot.·RESULTS:The overexpression of β1-integrin-GFP fusion gene by RCE cells was successfully established.β1-integrin transfection significantly promoted the adhesion of RCE cells to Matrigel ( P < 0.05 ).β1-integrin overexpression also promoted the migration ability of RCE cells and induced FAK phosphorylation in them (P < 0.05).·CONCLUSION:These data suggest that overexpression of β1-integrin promotes the adhesion and migration of RCE cells and that the FAK pathway may play an important role in this process.

Objective To study the relationships between hepatocellular carcinoma (HCC) and the polymorphisms,promoter methylation,and expression of glutathione S-transferases P1 gene (GST) P1 gene.Methods Using methylation-special PCR (MSP),the methylated status of CpG islands of GSTP1 gene in tumor tissues of 53 HCC and its adjacent nontumor tissues were studied.The enzyme activities of GSTP1 were evaluated by ultraviolet colormetry.And using PCR-RFLP,the genetic polymorphisms of the GSTP1 genes of 74 healthy controls and 53 HCC patients were studied.Results The diffe-rences of the frequency of GSTP1 Ile/Ile,Ile/Val and Val/Val genotypes between HCC patients and the normal controls did not reach statistical significance (X~2=0.84,v=2,P=0.656).The frequency of methyla- tion of CpG islands of GSTP1 gene was significantly higher among the HCC tumor tissues when com- pared to the corresponding nontumor tissues (X~2=19.08,P＜0.001),and significantly higher in stageⅢ-Ⅳcases when compared to the stageⅠ-Ⅱcases (X~2=4.84,P=0.028).GSTP1 enzyme activities of cytoplasm in tumor cells were lower significantly than that in the adjacent nontumor tissues (t=2.49, P=0.014),and significantly higher in stageⅠ-Ⅱcases when compared to the stageⅢ-Ⅳcases (t= 2.31,P=0.025).On the other hand,the GSTP1 enzyme activities of cytoplasm in tumor cells with methylated status of GSTP1 gene were significantly lower than that in tumor cells with unmethylation (t=3.50,P=0.001).Conclusion GSTP1 inactivation via CpG island hypermethylation may contrib- ute to the pathogenesis of HCC.

Under the influence of Chinese medicine scientization, Chengjiang Acupuncture School, which was originated in the Republic of China (1912 - 1949), has also undergone a series of scientific practice. The Chengjiang School established scientific research, education and medical organizations such as the Research Society of China Acupuncture-Moxibustion, China Acupuncture-Moxibustion School, Training School of China Acupuncture-Moxibustion and Sanatorium of Acupuncture-Moxibustion. Zhenjiu Zazhi (Journal of Acupuncture-Moxibustion) was established, and monographs and textbooks such as Zhongguo Zhenjiu Zhiliaoxue (Therapeutics of Chinese Acupuncture-Moxibustion) and Zhongguo Zhenjiuxue Jiangyi (Lectures on Chinese Acupuncture-Moxibustion) were published. Meanwhile, based on clinical practice, large members of acupuncture personnel with scientific literacy emerged. Therefore, it is held that the scientization of Chengjiang School has performed a profound influence on the development of modern acupuncture-moxibustion.
Acupuncture ; education ; history ; Acupuncture Therapy ; history ; China ; History, 20th Century ; History, 21st Century ; Humans ; Moxibustion ; history ; Schools ; history

Designing of natural product-like compounds using natural products as template structures is an important strategy for the discovery of new drugs. Gambogic acid (GA), which is a Garcinia natural product with a unique caged xanthone scaffold, inhibits potent antitumor activity both in vitro and in vivo. This review summarized the researches on the identification of the antitumor pharmacophore of GA, and the design, structural optimization and structure-activity relationship (SAR) of natural product-like caged xanthones based on it.
Antineoplastic Agents ; chemical synthesis ; chemistry ; pharmacology ; Biological Products ; chemical synthesis ; chemistry ; isolation & purification ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drug Screening Assays, Antitumor ; Garcinia ; chemistry ; Humans ; Molecular Structure ; Structure-Activity Relationship ; Xanthones ; chemical synthesis ; chemistry ; isolation & purification ; pharmacology

OBJECTIVETo investigate the efficacy of Chinese medicine Huangban Bianxing Recipe (HBR) for treatment of choroidal neovascularization (CNV).

METHODSEighty patients (97 eyes) suffered from CNV were assigned to two groups, the treated group treated by HBR orally taken one dose every day and the control group treated with ginkgo leaf tablet 1 tablet (19.2 mg) thrice a day, all for 3 months. The best corrected visual acuity (BCVA), fundus hemorrhagic area were measured; the neo-genetic vascular leakage area determined by fluorescent fundus angiography (FFA); and the out-layer high reflective band thickness (OHRBT) estimated by optical coherent tomography (OCT) were recorded before and after treatment.

RESULTSAfter treatment, in the treated group, BCVA was improved, with an effective rate of 67.12%, while in the control group, it was 33.33%, the former was better than the latter significantly (P < 0.05). The effective rate in reducing fundus hemorrhagic area in the two groups (85.11% vs 47.62%) and that in the CNV closure showed by FFA (67.50% vs 37.50%) were also superior in the treated group (P < 0.05). Besides, OCT showed that the OHRBT reduced significantly after treatment in the treated group (P < 0.01).

CONCLUSIONHBR can improve the vision, reduce the CNV leakage in the macular region, and advance the absorption of fundus hemorrhage in CNV patients.

Adult ; Aged ; Aged, 80 and over ; Choroidal Neovascularization ; drug therapy ; etiology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Macular Degeneration ; complications ; drug therapy ; Male ; Middle Aged ; Phytotherapy ; Visual Acuity ; drug effects ; Young Adult

· AIM: To investigate the effect of β1-integrin overexpression on the apoptosis of rabbit corneal epithelial cells and the related mechanism. · METHODS: The plasmid expressing β1-integrin-GFP fusion protein was constructed by polymerase chain reaction (PCR), and this plasmid (β1 group) or the empty vector (mock group) was transfected into rabbit corneal epithelial cells, respectively. The expression of β1-integrin-GFP fusion gene was confirmed by reverse transcription-polymerase chain reaction (RT-PCR) and Western blot. The adhesion of transfected cells to extracellular matrix (ECM) proteins was determined by adhesion assay. The apoptosis of rabbit corneal epithelial cells was assayed by Hoechst 33342 staining and DNA ladder. The phosphorylation of mitogen-activated protein (MAP) kinase was examined by Western blot. · RESULTS: Rabbit corneal epithelial cells overexpressing β1 -integrin-GFP fusion gene were successfully established. Compared with mock group, β1-integrin transfection significantly promoted the adhesive of rabbit corneal epithelial cells to ECM proteins such as laminin, fibronectin, collagen Ⅰ and collagen Ⅳ. Β1-integrin overexpression inhibited apoptosis and induced MAP kinase phosphorylation in rabbit corneal epithelial cells (P<0.05).· CONCLUSION: These data suggest that overexpression of β1-integrin confers resistance to apoptosis in rabbit corneal epithelial cells, and MAP kinase pathway may play an important role in this process.

Objective To construct multiple myeloma mucin MUC1-2VNTR gene eukaryotic expressing vector,which provided the basic material for further study of multiple myeloma DNA vaccine.Methods MUC1-2VNTR coding gene as target gene,and a KOZAK sequence was inserted before it.Hind Ⅲ and Xba Ⅰ restriction enzyme site were inserted on both ends.Then the whole sequence was synthesized and cloned into pcDNA3.1/myc-his B vector,and the recombinant vector was identified by restriction enzyme digestion and DNA sequencing.Results Synthesized MUC1-2VNTR gene was 140 bp.Restriction enzyme digestion and DNA sequencing confirmed pcDNA3.1/MUC1-2VNTR/myc-his B including the whole exact translation frame region and MUC1-2VNTR gene.Condnsion The pcDNA3.1/MUC1-2VNTR/myc-his B has been successfully constructed,which provides the basic material for further studies of MUC1 mucin function and multiple myloma DNA vaccine.

Objective To evaluate the clinical effect of popliteal artery perforator-based sural neurovascular flap for repairing soft tissue defects in middle and distal upper leg.Methods Between December,2011 to September,2015,18 cases with skin soft tissue defects on the middle and distal upper leg were treated with popliteal artery perforator-based sural neurovascular flap,in which 12 cases were males,and 6 cases were females.The age was from 24 to 55 years,with the average age of 35.9 years.The size of tissue defects ranged from 3.0 cm × 3.0 cm to 7.0 cm × 9.0 cm.Results All flaps survived completely in 18 cases,1 case of marginal infection heated after dressing,and the rest cases in Ⅰ healing.The outline and function of survived flap were satisfactory during 3-16 months follow-up,with two-point discrimination of 4.0-10.0 mm.Conclusion The anterograde sural neurovascular flap based on the popliteal artery perforator provides a practical option for covering tissue defects in middle and distal upper leg.This flap is characteristiced by reliable blood supply without sacrificing main vesses,good contour and texture,in addition,the operation is easy of handling.

Objective To explore the influence of intravenous injection combined with oral of metoprolol tartrate on left ventricular function and adverse cardiovascular events of patients with acute anterior myocardial infarction.Methods84 cases of Patients with acute anterior myocardial infarction treated in the First Hospital of Hebei Medical University were selected as the study objects, and were divided into vein group and combination group according to drugs-taking modes, 42 cases in each groups.The vein group were treated with intravenous injection of metoprolol tartrate, and the combination group were treated with intravenous injection combined with oral of metoprolol tartrate.Clinical effect, left ventricular function, BP and HR levels, and incidence of adverse cardiovascular events were observed in the two groups.ResultsThe total effective rate of the combination group was 95.24% significantly higher than that of 80.95% in the vein group(P<0.05).After treatment, LVEF was significantly higher than that before treatment in the two groups (P<0.05), which of the combination group was significantly higher than that of the vein group (P<0.05).The levels of LVESD and LVEDD were significantly lower than those before treatment in the two groups (P<0.05), which of the combination group were significantly lower than those in the vein group(P<0.05).After treatment, DBP, SBP and HR were significantly lower than those before treatment in the two groups (P<0.05), which of the combination group were significantly lower than those in the vein group(P<0.05).The incidence of adverse cardiovascular disease was 11.90% in the combination group, and it was no significantly different from that of 16.67% in the vein group (P>0.05).ConclusionIntravenous injection combined with oral of metoprolol tartrate can effectively improve left ventricular function of patients with acute anterior myocardial infarction, and reduce incidence of adverse cardiovascular events.