The animals have made huge contribution for the medical research of human life, and researching on animal welfare is crucial for developing biomedicine and enhancing human health. However, during the research of medical experiment, abusing laboratory animals happens a lot, many of them have suffered great damage and pain, even lose lives. Consequently, the legal issues related to the protection of laboratory animals become the focus con-cerned by people. Through analyzing the legislation status and existing problems in our country, elaborating the rel-evant legal institutions in foreign countries, as well as using the successful legislative experiences from abroad, this paper puts forward the countermeasures and suggestions for the laboratory animal welfare act that accords with our national condition.

Objective To evaluate the curative effects of venlafaxine in patients with depressive disorder between therapeutic modes of inpatient and outpatient. Methods Seventy-two patients with depressive disorder including 36 inpatients and 36 outpatients were measured respectively with HAMD24 after treatment for 1 week, 2 weeks, 4 weeks and 8 weeks, and then the curative effects were compared between two groups. Results It was found that all patients in the inpatient group completed the observation and two patients in the outpatient group didn’t complete the observation. The HAMD24 score was significantly decreased in the inpatient group than that of the outpatient group from the second week to the end of the eighth week (P<0.05). The total effective rates were 80.6% for inpatient group and 64.7% for outpatient group, no significant difference between them. Conclusion Venlafaxine shows a higher therapeutic value and outcome in the therapeutic mode of inpatient.

Abstract: Objective　To investigate the influences of notoginsenoside R1 (NGR1) on cell injury and Janus kinase 2 (JAK2)/signal transducer and activator of transcription 3 (STAT3) signaling pathway of alveolar epithelial cells infected by Klebsiella pneumoniae (Kp). Methods　A549 cells were grouped into five groups: control group (C group), infection group (Infect group), infection + low NGR1 group (Infect + L-NGR1 group), infection + high NGR1 group (Infect + H-NGR1 group), and infection+high NGR1+JAK2/STAT3 pathway inhibitor group (Infect+H-NGR1+SD-1029 group). Cell proliferation was measured using CCK8; ELISA kits were applied to detect the contents of interleukin-1β (IL-1β), tumor necrosis factor-α (TNF-α) and interferon γ (IFN-γ) in the culture medium; flow cytometry was applied to detect apoptosis; RT-qPCR was applied to detect the expressions of JAK2/STAT3; Western blot was applied to detect JAK2/STAT3 pathway, autophagy protein microtubule-associated protein 1 light chain 3 (LC3), autophagy-relatedgene5 (Atg5), autophagy-related gene (Atg) 6 (Beclin-1), apoptosis protein B-cell lymphoma 2 (Bcl-2), Bcl-2-accociated protein (Bax), cysteinyl aspartate specific proteinase (cleaved-caspase-3) proteins expression. Results　Compared with the C group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect group were obviously decreased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Compared with Infect group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the mRNA relative expressions and protein phosphorylation levels of JAK2, STAT3 in the Infect+L-NGR1 group and Infect+H-NGR1 group were obviously increased (P<0.05); the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-Caspase-3 were obviously decreased (P<0.05). Compared with Infect+H-NGR1 group, the 72 h cell viability, the protein levels of Bcl-2, LC3-II/I, Atg5, Beclin-1, the protein phosphorylation levels of JAK2, STAT3 in the Infect+H-NGR1+SD-1029 group were obviously decreased (P<0.05), and the contents of IL-1β, TNF-α, IFN-γ, apoptosis rate, the protein levels of Bax and cleaved-caspase-3 were obviously increased (P<0.05). Conclusions NGR1 can activate the JAK2/STAT3 signaling pathway, promote autophagy of alveolar epithelial cells, and inhibit Kp-induced inflammatory injury and apoptosis of alveolar epithelial cells.

Objective To investigate the therapeutic effect of Tiaozhi Decoction for the treatment of coronary heart disease induced stable angina complicated with hyperlipemia, and to observe its effect on serum inflammatory factors. Results One hundred qualified patients were evenly randomized into treatment group and control group. Both groups were given conventional western medical treatment with reference to Guide for Diagnosis and Treatment of Chronic Stable Angina, and additionally, the control group was given oral use of Simvastatin and the treatment group was given Tiaozhi Decoction orally. The treatment of the two groups covered 8 weeks. Before and after treatment, blood lipid levels of total cholesterol ( TC) , triglyceride ( TG) , low-density lipoprotein cholesterol ( LDL-C) , high- density lipoprotein cholesterol ( HDL-C) , apolipoprotein A (ApoA) and apolipoprotein B (ApoB) were observed. The frequency of angina pectoris attack and the dosage of Nitroglycerin Tablets per week in both groups were recorded during the treatment. Therapeutic effect on lowering blood lipid and on improving electrocardiogram was evaluated after treatment. Serum levels of hypersensitivity C-reactive protein (hs-CRP), interleukin-6 (IL-6), homocysteic acid (Hcy), adiponectin (APN), and oxidized low-density lipoprotein (ox-LDL) were detected before and after treatment. Results(1) The results of Ridit analysis showed that the treatment group had better therapeutic effect on lowering blood lipid and on improving electrocardiogram than the control group ( P<0.05) . ( 2) After treatment, TG, HDL-C, ApoB and ApoA levels were much improved in the treatment group compared with those in the control group (P<0.01) . ( 3) The frequency of angina pectoris attack and the dosage of Nitroglycerin Tablets per week were reduced in the treatment group compared with those in the control group (P<0.01) . (4) After treatment, the treatment group had lower hs-CRP, IL-6, Hcy and ox-LDL levels, and higher APN level than the control group (P<0.01) . Conclusion Tiaozhi Decoction is effective for the treatment of coronary heart disease induced stable angina complicated with hyperlipemia through improving lipid metabolism and reducing angina pectoris attack, and the mechanism may be related with the regulation of inflammatory factors.

Objective To evaluate the efficacy and safety of oral ginkgo biloba extract EGB761 in patients with mild cognitive impairment.Methods They searched PubMed,Embase,the Cochrane Library ,CNKI,Wanfang and VIP databases for randomly controlled trials of oral ginkgo biloba extract for mild cognitive impairment.After assessed the quality of studies included,RevMan5.2 software was used to analyze data.Results Seven studies which including 815 patients were involved by our inclusion criteria.The results of meta-analysis showed,compared with the control group,ginkgo biloba was superior in improving mild cognitive impairment patients' MMSE level[MD=1.81,95%CI(0.02,3.60),P=0.05;MD=1.96,95%CI(1.48,2.43),P<0.000 01;MD=1.79,95%CI(0.99,2.58),P<0.000 1] after treated three months、six months and twelve months.Ginkgo biloba was also superior in improving mild cognitive impaimant patients.CDT level[MD=0.43,95%CI(0.30,0.57),P<0.000 01;MD=0.57,95%CI(0.39,0.75),P<0.000 01] after treated six months and twelve months.The effect of preventing MCI patients into dementia was better than that of the control group[RR=0.27,95%CI(0.06,1.27),P=0.10;RR=0.32,95%CI(0.16,0.63),P=0.001]after treated six months and twelve months.Conclusion Oral preparation of ginkgo biloba extract in the treatment of MCI clinical efficacy and prevention of dementia occurrence rate was better than that of blank control group.

Transcatheter arterial chemoembolization (TACE) has already been a mature and an effective treatment for advanced hepatocellular carcinoma (HCC).Clinically,it is very important to quickly and accurately evaluate the postoperative curative effect with minimally invasive technique so as to determine the next treatment options.At present,postoperative conventional CT and MRI are the main means to assess the curative effect of TACE,but it is a pity that after the treatment the functional changes of the tumor occur earlier than the morphological changes.In recent years,functional MRI techniques,such as diffusionweighted imaging (DWI),multi-b value DWI,dynamic contrast-enhanced (DCE) imaging,etc.have been more and more used for quantitative evaluation of the diffusion of water molecules and the blood microcirculation perfusion within the tumor tissue,and some progresses have been achieved in the evaluation of curative efficacy for tumor.This paper aims to make a comprehensive review about the research progress of the above mentioned functional imaging methods as well as their current application status in evaluation of the curative effect of TACE.

Background It has been proved that as an important adhesion protein of extracellular matrix,osteopontion (OPN) can affect tumor neovascularization.Some new researches showed that anti-OPN antibody plays a role in regulating the neovascular vessel formation.Choroidal neovascularization (CNV) has the same structure with tumor neovascularization,but whether anti-OPN antibody restricts new vessel formation is unclear.Objective This study was to investigate the inhibitory effect of anti-OPN antibody on CNV.Methods Laser-induced CNV models were created in 40 eyes of 40 male SPF C57BL/6J mice by Argon laser photocoagulation of retinas,with the wavelength 514 nm.Thirty-six successful models were randomly divided into anti-OPN antibody group,mouse-IgG group and PBS group by the randomized number table.On the second day after photocoagulation,anti-OPN antibody of 400 μg was intraperitoneally injected in the anti-OPN antibody group,and the equivalent amount of mouse IgG and PBS were used in the same way in the mouse IgG group and PBS group.The CNV was evaluated by fundus fluorescein angiography (FFA) on the seventh days after photocoagulation.The mice were immediately sacrificed and the eyeballs were enucleated on the fourteenth day after photocoagulation,and 4 eyeballs in each group were used to observe the areas of CNV on the retinal pigmental epithelium-choroid-sclera fiat mounts,and the other 8 eyeballs of each groups were used to analyze the expression levels of OPN mRNA and vascular endothelial growth factor(VEGF) mRNA using quantitative fluorescence-PCR (QF-PCR).Results FFA showed fluorescein leakage areas around laser spots 7 days after photocoagulation,indicating that CNV appeared.The CNV areas were ([16.98±0.70] × 103) μm2,([27.13 ± 0.81] × 103) μm2 and ([35.39±2.14] ×103) μm2 respectively in the anti-OPN antibody group,mouse IgG group and PBS group,with a significant difference among the 3 groups (F =533.76,P =0.00),and the CNV area was significantly smaller in the anti-OPN antibody group compared with those of the mouse IgG group and PBS group (q =-3.95,-4.40,both at P＜0.05).No significant difference was found in the OPN mRNA expression between the antiOPN antibody group and mouse IgG group (t =-5.26,P =0.66).However,the expression of VEGF mRNA in choroidal tissue was significantly declined in the anti-OPN antibody group than that in the mouse IgG group (t =-6.74,P＜0.01).Conclusions Anti-OPN antibody suppresses the formation of CNV in laser-induced mouse model by down-regulating VEGF.

ObjectiveTo investigate the risk factors of preterm birth, as well as the clinical characteristics in dichorionic diamniotic (DCDA) twins and monochorionic diamniotic (MCDA) twins. MethodsA retrospective study was conducted on 290 premature cases out of 363 twin pregnancies who delivered alive babies in the First Affiliated Hospital, Sun Yat-sen University from September 2012 to March 2015. The selected cases, including 219 cases of DCDA and 71 cases of MCDA,were divided into three groups according to their gestational age at delivery: 28-31+6, 32-33+6 and 34-36+6 weeks. The clinical features, causes and risk factors were described between these three groups. Analysis of variance,Chi-square test and multi-variant Logistic regression were used for statistical analysis.ResultsThe incidence of premature delivery in twin pregnancies was 79.9% (290/363), while this figure was lower in DCDA twins than in MCDA [76.3%(219/287) vs 93.4%(71/76),χ2=10.955,P=0.001]. The three leading causes of preterm birth in DCDA twins were gestational age≥36 weeks (33.8%, 74/219), preterm labor (30.6%, 67/219) and preterm premature rupture of membrane (PPROM) (8.7%, 19/219), while in MCDA twins were preterm labor (31.0%, 22/71), selective intrauterine growth restriction (21.1%, 15/71) and gestational age≥36 weeks (19.7%, 14/71). Logistic regression analysis showed that the independent risk factors of preterm birth in twins at 28-31+6 weeks was PPROM (OR=2.390, 95%CI: 1.006-5.872,P=0.043), and for those twins at 32-33+6 weeks, the independent risk factors were MCDA (OR=2.758, 95%CI: 1.243-6.118,P=0.013), preeclampsia (OR=12.176, 95%CI:4.685-31.642,P=0.000), PPROM (OR=5.348, 95%CI: 2.151-13.294,P=0.000) and preterm labor (OR=3.274, 95%CI:1.453-7.375,P=0.004). MCDA (OR=3.666, 95%CI: 1.364-9.585,P=0.010) and preeclampsia (OR=8.086, 95%CI:1.044-62.617,P=0.045) were the risk factors in the group of 34-36+6 weeks.ConclusionsAlthough preterm birth in MCDA and DCDA twins is due to different reasons, the former has a higher incidence than the latter. The risk factors of premature delivery at different gestations are also different.

Objective To obtain recombinant human stanniocalcin 1 ( STC1 ) with biological activity in Escheri.coli cells expression.Methods The gene was cloned into pET32b( +) vector by fused with thioredoxin and His tag .E.coli BL21(DE3) competent cells were transfomed by the recombinant vector .After renaturation, the fusion protein was digested with thrombin and intact STC1 protein was purified from the digested protein using Ni ion affinity chromatography .Recombi-nant humanSTC1 protein was confirmed by Western blot analysis using goat anti-STC1 antibody.The biological activity of STC1 in rat was assayed using standard method for assessment of renal function .Results The recombinant human STC 1 fu-sion protein is successfully expressed in Escherichia coli, the fusion protein was purified by affinity chromatography from the inclusion body and renaturated .Intact hSTC1 protein was released by thrombin digestion and purified by Ni ion affinity col-umn.The intact STC1 proteins was confirmed by Western blot analysis .Rat bioassay revealed that STC1 boosted phosphate reabsorption.Conclusion Recombinant STC1 protein was successfully expressed and has native biological activities .This protein could be used as an antigen for the preparation of monoclonal antibody against humanSTC 1.

Objective To investigate the influence of prepubertal exposure to estradiol benzoate (EB)in the male reproductive system of the rats and the natural process of tissue repair,and to clarify the possible mechanism of the reproductive toxicity of exogenous estrogen.Methods Ninety 2 1-day-old male Wistar rats were randomly divided into 2 experimental groups (low dose of EB group and high dose of EB group,n=30)and control group (n=30). The rats in the experimental groups were injected with EB dissolved in peanut oil at 15(low dose of EB group)and 15 000μg·kg-1 (high dose of EB group)respectively,the rats in control group received equal vehicle injection only,once every other day for two weeks from postnatal day(PND)21 to 34.All of them were normally fed after the drug usage was stopped.The testes were harvested at the stages of PND 60 and PND 125(n=15 at each stage).The serum levels testosterone of (T),follicle-stimulating hormone(FSH),luteinizing hormone(LH),prolactin (PRL)and estradiol(E2)of the rats in various groups were detected with radioimmunology method and the weights of the rats in various groups were recorded;the histological changes of the testes tissue were observed with light microscope.Results On PND60,compared with control group,the T levels in low dose of EB group and high dose of EB group were decreased(P<0.05 or P<0.01);the FSH,LH and E2 levels were increased (P<0.05 or P<0.01),and the PRL levels had no change(P>0.05);the weights of testes were decreased(P<0.01);the histological changes of the testes of the rats in experimental groups included seminiferous tubules maldevelopment, decreased cell number of seminiferous epithelia.Compared with low dose of EB group,the T and FSH levels in high dose of EB group were decreased (P<0.05 or P<0.01),the E2 and LH levels were increased(P<0.01),the PRL level had no change (P>0.05 ), and the weight of testes was decreased (P<0.01 );the diameters of seminiferous tubules were smaller,there was no sperm in high dose of EB group while there were a few sperms in low dose of EB group.On PND125,compared with control group,the T,FSH and PRL levels in low dose of EB group and high dose of EB group were decreased(P<0.01),the E2 levels were increased (P<0.01);the LH level in low dose of EB group was increased(P<0.05),the LH level in high dose of EB group was decreased(P<0.01), and the weights of testes in high dose of EB group were decreased(P<0.01);the diameters of seminiferous tubules and the cell number of seminiferous epithelia were increased but not apparent change.Compared with low dose of EB group,the T,LH levels in high dose of EB group were decreased (P<0.01),the E2 and FSH levels were increased(P<0.01),the PRL level had no change(P>0.05),and the weight of testes was decreased(P<0.01);there was still no sperm in high dose of EB group, the number of sperms was increased in low dose of EB group, but it was still lower than that in control group.Conclusion EB is harmful to the reproductive system and can change the normal serum sex hormone levels,even induces the irreversible injury.