Objective: To investigate the relationship between G505A polymorphism in the encoding region of thrombin-activated fibrinolysis inhibitor (TAFI) and cerebral infarction in Chinese Han population. Methods: A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was used to detect G505A polymorphism in the encoding region of TAFI in 130 patients with cerebral infarction and 118 healthy individuals. Results: The GG genotype of TAFI G505A accounted for 35.4% (46/130) and GA or AA genotype accounted for 64.6% (84/130) in the cerebral infarction group; and they were 49.2% (58/118) and 50.8% (60/118) respectively in the control group. The difference was statistically significant (P = 0.028). The G and A allele frequencies were 60.4% (157/260) and 39.6% (103/260) respectively in the cerebral infarction group, and they were 69.9% (165/236) and 30.1% (71/236) respectively in the control group. The difference was statistically significant (P = 0.026). Multivariate logistic regression analysis showed that G505A polymorphism in the encoding region of TAFI was an independent risk factor for cerebral infarction (OR = 2.660, 95% CI 1.330-5.317, P = 0.006). Conclusion: The TAFI G505A polymorphism may be one of the risk factors for cerebral infarction.

Objective To express human thyroid peroxidase (hTPO) epitopes gene and apply its products in clinical assay. Methods hTPO epitopes gene was cloned into expression vector pGEX 4T 3 then transformed into E. coli BL21. Expression of hTPO gene was induced by isopropyl ? D thiogalactoside, expressed product (GST hTPO) was purified by affinity chromatography and their immunoactivity was demonstrated. ELISA technique using GST hTPO as antigen was established for determining TPOAb. Serum TPOAb level was determined, and HLA DR antigen, dendritic cells and lymphocytes in the thyroid gland tissue were observed in these same AITD patients.Results GST hTPO acquired from procaryotic expression had high purity and good immunoactivity. The CVs of the ELISA technique established with GST hTPO were between 5.93%～7.59%. A significantly positive correlation was found between the TPOAb levels determined respectively by ELISA and RIA method. Serum TPOAb level and distribution of HLA DR antigen and dendritic cells showed the same ascending tendency following the aggravation of lymphocyte infiltration. Conclusion Product of genetic engineering, GSH hTPO, can be used to establish a clinical assay for TPOAb. The correlation between the level of serum TPOAb and the detriment of thyroid tissue is demonstrated.

The effects of betulinic acid (BA), a pentacyclic lupane-type triterpene, on the cell viability, cell cycle and apoptosis in human leukemia K562 cells were investigated. The effects of BA on the growth of K562 cells were studied by MTT assay. Apoptosis was assayed through Annexin V/propidium iodide (PI) double-labeled cytometry. The effects of BA on the cell cycle of K562 cells were studied by a PI method. The expression of Bax and capase-3 was detected by using Western blot. The results showed that BA was cytotoxic to K562 cells with an IC(50) of 21.26 mug/mL at 24 h. After treating K562 cells with 10 mug/mL BA for 72 h, the number of cells was reduced by 58%. BA induced apoptosis of K562 cells in a time- and dose-dependent manner. The proportion of cells in G(0)/G(1) and G(2)/M phases was decreased and that in S phase was increased after K562 cells were treated with BA for 24 h. BA treatment also increased the expression of the pro-apoptotic proteins Bax and caspase-3. It suggested that BA could inhibit the proliferation of K562 cells through the induction of cell cycle arrest and apoptosis. The antitumor effects of BA were related with up-regulation of the expression of Bax and caspase-3 proteins. BA may qualify for the development of new therapies for leukemia.

The isolated 24 strains-producing hydantoinase & carbamoylase were first identified by Biolog microbial identification system and 16S rDNA sequence analysis.The results suggested that the hydantoinase & carbamoyalse-producing bacteria belonged to Bacillus,Geobacillus,Brevibacillus,Aneurinibacillus,Microbacterium,Pseudomonas,Kurthia and Empedobacter,and so on.Especially,Kurthia and Empedobacter were new hydantoinase & carbamoylase-producing genera.Furthuremore,it was found that D-hydatoinase & carbamoyalse-producing bacteria belonged to Pseudomonas and Agrobacterium,while most of L-hydantoinase & carbamoyalse-producing bacterial belonged to Bacillus,Geobacillus and Microbacterium.The distribution feature of D-hydantoinase & carbamoyalse-producing bacteria and L-hydantoinase & carbamoyalse-producing bacteria showed some genera tendency.This research work will provide the biomaterial of different hydantoinase and carbamoylase and contribute to study the structure and function,molecular evolution of the two enzymes.

OBJECTIVETo verify the regulatory effects of acupuncture on exercise tolerance in patients with chronic obstructive pulmonary disease (COPD) at stable phase.

METHODSThirty cases of COPD were randomly divided into a treatment group (16 cases) and a placebo group (14 cases). Based on specified aerobic exercise, acupuncture was applied in the treatment group and placebo acupuncture was used in the placebo group. The acupoints included Danzhong (CV 17), Rugen (ST 18), Guanyuan (CV 4), Zhongwan (CV 12), Tianshu (ST 25) and so on. The needle did not penetrate into the skin for the placebo group. The treatment was required for 2 to 3 times per week for totally 5 weeks. The indices of exercise tolerance, including 6-min walking distance (6-MWD), exercise time, maximum oxygen uptake (VO2max) forced expiratory volume in 1 second (FEV1)/forced vital capacity (FVC), maximum ventilatory volume (MVV), St. George respiratory questionnaire (SGRQ) were observed in two groups before and after treatment.

RESULTS(1) Exercise tolerance: the differences of 6-MWD and exercise time were statistically significant between groups, which were more superior in the treatment group (both P<0.01); the VO2max was significantly increased after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05). (2) Pulmonary ventilation function: the differences of FEV1%, FEV1/FVC and MVV% were statistically significant between groups, which were more superior in the treatment group (P<0.05, P<0.01); (3) SGRQ: the SGRQ was significantly improved after treatment in the treatment group (P<0.05), but there was no difference between two groups (P>0.05).

CONCLUSIONThe acupuncture could improve the exercise tolerance in patients with chronic obstructive pulmonary disease at stable phase, and shorten the onset time of aerobic exercise. Besides, acupuncture combined with aerobic exercise could effectively improve the pulmonary function.

Acupuncture Points ; Acupuncture Therapy ; Aged ; Exercise Tolerance ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Disease, Chronic Obstructive ; physiopathology ; therapy

Objective To investigate the effect of modified electroconvulsive therapy (MECT) with different anesthetics on efficacy of patients with treatment-resistant depression (TRD). Methods Ninety patients with TRD were enrolled in this study to receive a standard 8 times MECT. The HAMD-17 scale was evaluated before MECT and after the completion of the first, second, third, forth, sixth and eighth MECT session. The TESS scale was evaluated before MECT and after the completion of the last MECT session. Results Scores of HAMD-17 after the completion of the first, second, third, fourth, sixth and eighth MECT session were significantly reduced (P<0.05). There were significant differences of HAMD scales among the three groups since the first MECT session (P < 0.05). The remission rate of ketamine group, propofol group and mixed group was 96.7%, 43.3%, and 73.3% (P < 0.05). Conclusion MECT of ketamine anesthetic might contribute to the best effect of TRD.

Objective To construct a recombinant plasmid pGEX-TSO45W-4B-TSOL18 of Taenia solium,to obtain purified recombinant protein TSO45W-4B-TSOL18 and to prepare rabbit antiserum against the recombinant fusion protein.Methods TSO45W-4B and TSOL18 encoding genes were connected with hydrophobic (Gly4Ser)3 linker.TSO45W-4B-TSOL18 fusion gene was synthesized and cloned into an expression vector pGEX-1λT to construct a recombinant plasmid pGEX-TSO45W-4B-TSOL18.The positive recombinants were transformed into Escherichia(E.) coli ArcticExpress (DE3),and the expression of recombinant protein was induced with isopropylβ-D-thiogalactopyranoside (IPTG) and analyzed by sodium dodecyl sulfate polyacrylamide gelelectrophoresis (SDS-PAGE).The expressed recombinant fusion protein was purified through affinity chromatography.The purified recombinant protein was mixed with complete Freunds adjuvant at a dosage of 0.5 mg.Rabbit was intramuscularly and subcutaneously immunized in the hind leg and the back,respectively.After 2 weeks the rabbit was boosted with purified recombinant protein which was mixed with incomplete Freunds adjuvant at the same dosage.Rabbit can be boosted every 10 days until an adequate response was achieved.At the 6th day after the last immunization,blood was collected from the rabbit heart.Serum was separated to purify and prepare the antiserum.ELISA was applied to determine the titer of the antiserum and Western blot assay was used to determine the specificity of the antiserum.Results The size of the synthesized fusion gene TSO45W-4B-TSOL18 was 789 bp.The results of restriction enzyme digestion showed that the recombinant plasmid pGEX-TSO45W-4B-TSOL18 was successfully constructed.DNA sequencing showed that the size of the fusion gene TSO45W-4B-TSOL18 was 789 bp,which was consistent with expected result.As demonstrated by SDS-PAGE,relative molecular mass of the expressed recombinant fusion protein was approximately 55 × 103,and its purity was 85％ after purification with affinity chromatography.Titer of the antiserum against the recombinant protein was 1 ∶ 512 000 in ELISA assay and the specific rabbit antiserum against the purified recombinant protein TSO45W-4B-TSOL18 could specifically bind to the recombinant fusion protein in Western blot assay.The relative molecular mass of the specific band was about 55 × 103,which was consistent with expected size.Conclusions The recombinant plasmid pGEX-TSO45W-4BTSOL18 of Taenia solium is successfully constructed.High quality recombinant fusion protein TSO45W-4B-TSOL18 and high titer rabbit antiserum are successfully prepared.

OBJECTIVE:To study the anti-inflammatory effect of Xinfeng capsule on adjuvant arthritis (AA) in rats. METH-ODS:70 SD rats were randomized into a normal control group(isovolumetric normal saline),a model control group(isovolumet-ric normal saline),a positive control TCM group [Qufeng zhitong capsule 0.4 g(crude drug)/kg],a positive control chemical medi-cine group(Leflunomide tablet 2.1 mg/kg)and groups of low,middle and high doses of Xinfeng capsule,which were respectively marked as groups A,B,C,D,E,F and G. All groups of rats except for Group A were given freund’s complete adjuvant intracu-taneous injection to establishe AA models,and from the 12th day after the inflammation was induced,were given the correspond-ing drug,ig,for 28 consecutive days. The degree of toe swelling and arthritis index were determined and calculated for all groups of rats before and after the administration. The contents of interleukin 1β(IL-1β) and tumor necrosis factor α(TNF-α) in serum and synovial membrane of all groups of rats were determined 24 h after the last administration,and pathomorphological changes of their ankles were observed by light microscope. RESULTS:Compared to group A,group B demonstrated higher degree of toe swelling,arthritis index and contents of IL-1β and TNF-α in serum and synovial membrane,with statistical significance(P<0.01). After the administration,the degree of toe swelling and arthritis index of groups C,D,E,F and G reduced,with statistical signifi-cance(P<0.05). Compared to group B,groups C,D,E,F and G showed lower degree of toe swelling,arthritis index and con-tents of IL-1β and TNF-α in serum and synovial membrane,with statistical significance(P<0.01 or P<0.05). Synovial membrane hyperplasia and inflammatory cell infiltration were noted in the ankle tissue of the rats in group B,and such symptoms were not so serious in groups C,D,F and G. CONCLUSIONS:Xinfeng capsule has anti-inflammatory effect for AA rats by a mechanism which may be related to accelerating synoviocyte apoptosis and inhibiting synovial membrane hyperplasia.

Objective To study the clinical characteristicsand perioperative managementof complicated placenta increta, effectively reduce the maternal adverse perinatal outcomes. Methods Retrospective analysis 25 cases of complicated placenta increta between January 2013 and December 2015 in the Third Affiliated Hospital Of Guangzhou Medical University. Grouped into preoperative line 9 cases of ureteral catheter group and without catheter group 16 cases; Conventional hysterectomy group of 17 cases and the posterior hysterectomy group of 8 cases , compare the operation time , postpartum hemorrhage , blood transfusion amount , bladder injury or ureteral injury rate , rate of transferred to the ICU and hospital stay. Results 76% appear repeatedly painless vaginal bleeding during pregnancy , 56% appear bleeding before delivery. Prenatal diagnosis of 17 cases (68%). The preoperative line cystoscopy + bilateral retrograde ureteral catheter or after the posterior hysterectomy , shorter operation time , less postpartum hemorrhage , reduce blood transfusion volume , no urinary tract injury rate, transferred to the ICU rate is low, the difference was statistically significant (P < 0.05). Conclusions We should attach importance to repeated painless vaginal bleeding , improve prenatal diagnostic rate of complicated placenta increta. The perioperative managementis more comprehensive , effective and standard participation , preoperative ureteral catheter and the posterior hysterectomy can effectively reduce the maternal adverse perinatal outcomes.

OBJECTIVE To investigates the effects of imperatorin on the oxidative stress in the cerebral cortex and hippocampus after focal cerebral ischemia/reperfusion injury. METHODS Transient focal cerebral ischemia/reperfusion model in male Sprague-Dawley rats was induced by 2 h middle cerebral artery occlusion followed by 24 h reperfusion. Imperatorin (1.25 and 2.5 mg·kg- 1) or vehicle were administered intraperitoneally at 1, 5 and 9 h after the onset of ischemia. At 24 h after reperfusion, the biomarkers of oxidative stress such as the levels of reactive oxygen species (ROS), lipid peroxidation products malondialdehyde (MDA), nitric oxide (NO) and total antioxidant capacity (T-AOC), the activities of inducible nitric oxide synthase (iNOS), superoxide dismutase (SOD) and catalase (CAT) in the cerebral cortex and hippocampus were observed. We also assessed the nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1), and the NAD(P)H-quinone oxidoreductase 1 (NQO-1) protein expression by Western blot. RESULTS As compared to vehicle-treated animals, imperatorin treatment significantly reduced the ROS, MDA, NO levels and iNOS activity, increased T-AOC and the activities of SOD and CAT. Furthermore, imperatorin treatment also significantly induced the nuclear translocation of Nrf2, enhanced the protein expression of HO-1 and NQO-1 in the cerebral cortex and hippocampus. CONCLUSION Our findings indicate that imperatorin can protect the brain against the excessive oxidative stress induced by cerebral ischemia/reperfusion through activation of Nrf2 signaling pathway.