Governor vessel moxibustion combined with western medication for mild to moderate depression with kidney-yang deficiency: a randomized controlled trial / 中国针灸

OBJECTIVE@#To compare the therapeutic efficacy of governor vessel moxibustion combined with fluoxetine hydrochloride capsule, simple fluoxetine hydrochloride capsule and placebo moxibustion combined with fluoxetine hydrochloride capsule for mild to moderate depression with kidney-yang deficiency.@*METHODS@#A total of 126 patients with mild to moderate depression with kidney-yang deficiency were randomized into a governor vessel moxibustion group (42 cases, 2 cases dropped off), a western medication group (42 cases, 1 case dropped off) and a placebo moxibustion group (42 cases, 1 case dropped off). The western medication group was given fluoxetine hydrochloride capsule orally, 20 mg a time, once a day. On the basis of the treatment in the western medication group, governor vessel moxibustion was applied from Dazhui (GV 14) to Yaoshu (GV 2) in the governor vessel moxibustion group, once a week; placebo moxibustion was applied in the placebo moxibustion group, once a week. Treatment of 8 weeks was required in the 3 groups. Before and after treatment, the scores of Hamilton depression scale-17 (HAMD-17), Asberg's rating scale for side effects (SERS) and TCM clinical symptom were compared, and the clinical efficacy was evaluated.@*RESULTS@#After treatment, the scores of HAMD-17, SERS and TCM clinical symptom were decreased compared before treatment in the 3 groups (P<0.05), the decrease ranges of above scores in the governor vessel moxibustion group were larger than those in the western medication group and the placebo moxibustion group (P<0.05). The total effective rate was 92.5% (37/40) in the governor vessel moxibustion group, which was higher than 75.6% (31/41) in the western medication group and 80.5% (33/41) in the placebo moxibustion group (P<0.05).@*CONCLUSION@#Governor vessel moxibustion combined with fluoxetine hydrochloride capsule can improve the degree of depression and relieve the clinical symptoms in mild to moderate depression patients with kidney-yang deficiency, the efficacy is superior to simple fluoxetine hydrochloride capsule, and can reduce the fluoxetine hydrochloride capsule-induced adverse effect to a certain extent.

LC-MS fingerprint and multi-indicator components analysis of classical formula Gualou Xiebai Banxia Decoction / 中国中药杂志

This study developed an optimal pre-processing technique for the reference substance of the classic formula Gualou Xiebai Banxia Decoction(GXBD) and established a comprehensive quality control method for GXBD reference substance to provide a reference for its overall quality evaluation. The authors prepared 15 batches of GXBD samples and innovatively used the extracted ion chromatogram under the base peak chromatogram mode to establish a liquid chromatography-mass spectrometry(LC-MS) fingerprint, identify characteristic peaks, and perform quantitative analysis of indicator components. The yield of the 15 batches of GXBD samples ranged from 50.28% to 76.20%. In the positive ion mode, 12 common characteristic peaks were detected in the LC-MS fingerprint, and the structures of five common peaks were identified by comparison with reference standards. The similarity between the fingerprint profiles of different batches of samples and the reference fingerprint profile ranged from 0.920 to 0.984. Finally, liquid chromatography-triple quadrupole mass spectrometry(LC-QQQ/MS) in multiple reaction monitoring(MRM) mode was used to determine the content of eight indicator components in GXBD, including loliolide, chrysoeriol, rutin, cucurbitacin D, macrostemonoside Ⅰ, 25S-timosaponin B Ⅱ, 25R-timosaponin B Ⅱ, and peptide proline-tryptophan-valine-proline-glycine(PWVPG). The method established in this study can reduce matrix interference in the compound, and it has good accuracy, stability, and practical value. It effectively reflects the quality attributes of GXBD samples and can be used for the comprehensive quality control of GXBD.

Location Analysis of Volatile Oil in Angelicae Sinensis Radix by Frozen Section and Fluorescence Imaging / 中国实验方剂学杂志

ObjectiveTo establish a simple， fast and accurate method for locating the volatile oil in Angelicae Sinensis Radix based on frozen section and fluorescence imaging technology， and to reveal the distribution and accumulation of volatile oil in the roots of this herbal medicine. MethodAngelicae Sinensis Radix was used as the research material， the best frozen section conditions for the research material were established by comparing the effects of different cryoprotectants on the quality of frozen sections of Angelicae Sinensis Radix. The suitability of Sudan Ⅲ chemical staining and fluorescence localization for positioning the volatile oil were compared according to the loss of volatile oil and the complexity of operation process. ResultA new method for evaluating the quality of frozen sections of Angelicae Sinensis Radix was established. According to the evaluation equation， it was found that the highest score was obtained when the head， body and tail positions of Angelicae Sinensis Radix were treated with 20% glycerol， 15% glycerol and 20% sucrose， respectively. There was yellowish-brown oily substance in the oil chambers of phelloderm and secondary phloem， and oil canal of the secondary xylem of Angelicae Sinensis Radix， which could be stained orange red or orange yellow by Sudan Ⅲ， and there was green spontaneous fluorescence in the same part under the fluorescence microscope. ConclusionThe relatively complete section of Angelicae Sinensis Radix can be obtained after being treated with cryoprotectant. The volatile oil exists in the oil chambers of phelloderm and secondary phloem， and oil canal of the secondary xylem of Angelicae Sinensis Radix. This study can provide reference for observation of the accumulation sites of volatile oil in other plants.

Comparison of clinical efficacy of two different pin arrangements for external fixation of femoral neck fracture / 中国骨伤

OBJECTIVE@#To retrospectively analyze the clinical efficacy of external fixation in the treatment of femoral neck fracture with two different pin layout.@*METHODS@#From April 2000 to April 2018, 140 cases of femoral neck fracture were treated with closed reduction and percutaneous pin external fixation, among them 121 cases were followed up for more than 1 year, including 31 cases in traditional group, 12 males and 19 females, aged 45 to 74(65.4±8.4) years;90 cases in modified group, 39 males and 51 females, aged 12 to 75 (64.5±7.8) years. In traditional group, the first needle was put on the femoral talus, the second and third needles were put under the tension line, and the three needles were not on the same line in the lateral phase; in modified group, the first needle was drilled into the lateralcortex of the femur, obliquely penetrating the distal and proximal end of the femoral talus fracture, and the other two needles were drilled into the medial cortex of the femoral neck and the femoral talus, respectively. The operation time, hospital stay, postoperative ambulation time, femoral neck shortening rate, fracture healing time, fracture healing rate and femoral head necrosis rate of the two groups were observed and compared. Harris hip function score was used one year after operation.@*RESULTS@#These 121 patients were followed-up, the follow up time of traditional group was 13 to 45(30.5±11.4) months;the follow-up time of modified group was 14 to 120(34.5±12.5) months. There was no significant difference in operation time, hospital stay and femoral head necrosis rate between two groups (@*CONCLUSION@#Compared with the traditional group, the modified group has the advantages of lower femoral neck shortening rate, shorter fracture healing time, higher fracture healing rate and higher Harris hip function score.

Prevalence of myopia among primary school students in mainland China：a Meta-analysis / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To estimate the pooled prevalence of myopia among primary school students in mainland China during 1980- 2013. Myopia had become a growing public health issue, with high prevalence rates in mainland China, particularly among children. However, we still had no population-based nationwide studies of the prevalence of myopia among primary school students in recent years. METHODS: Wanfang, Chinese National Knowledge Infrastructure and PubMed databases were searched independently until Dec. 31, 2013 to identify relevant articles. Data from the eligible articles were extracted by two reviewers. All of the data analyses were conducted using Meta-Analyst software (version 3. 13, USA). RESULTS: Thirty - seven eligible studies published between 1980 and 2013 were selected with a total of 245248 individuals. The pooled prevalence of myopia among the included individuals was 26. 5% (95% CI: 21. 8% -31. 7%). The prevalence of myopia increased with age (from 8. 4%at 6-8y to 57. 4% at 12-14y). CONCLUSION: The pooled prevalence of myopia among primary school students in mainland China was much higher than that of western countries or regions. The prevalence of myopia increased with age among primary school students. This study should be valuable for myopia prevention and treatment in mainland China.

Effect of asragaloside IV on vascular smooth muscle cells and neointima hyperplasia in rat carotid artery and their mechanism / 中草药

Objective: To investigate the effects of asragaloside IV (AS-IV) on cultured vascular smooth muscle cells (VSMCs) and neointima hyperplasia in the carotid artery of rats after ballon injury and explore the inhibitory mechanisms. Methods: For next related researches, primary cultures of VSMCs were prepared from the thoracoabdominal aorta of rats using explant method. Taking the recombinant rat tumor necrosis factor (TNF-α, 100 ng/mL ) as the stimulating factor, the model of VSMCs proliferation was established by TNF-α inducer in vitro and the effects of AS-IV (0, 0.5, 5, 25, and 50 μg/mL) on the VSMCs proliferation induced by TNF-α were determined by CCK-8 method. Rat carotid artery balloon injury model was prepared by Fogarty (2F) balloon catheter. Fifty healthy male Sprague-Dawley rats were randomly divided into five groups: a Sham-operation group (Sham), a model group (model), and three AS-IV-treated (20, 40, and 60 mg/kg) groups. Three days before the surgery, 1% CMC, AS-IV 20, 40, and 60 mg/kg were ig administered to each group once daily for continuous 17 d. Fifteen days after the surgery, rats were killed, and the carotid arterys were harvested. Hematoxylin-elsin staining was carried out to observe the pathomorphological change in vascular intima. The measurement of lumen area, intimal area, intimal area/medium film area were measured by computer image analysis system; Immunohistochemistry staining was performed to measure the expression of proliferating cell nuclear antigen (PCNA) and platelet-derived growth factor-BB (PDGF-BB). Results: The proliferative activity of VSMCs was obviously increased in the TNF-α group under the stimulation of TNF-α. There was a significant difference compared with the control group (P < 0.01). However, when pretreated with AS-IV ahead of time, we found that AS-IV significantly inhibited TNF-α-induced VSMCs proliferation in a dose- and time-dependent manner compared to the TNF-α group. Compared with the model group, the area of intima, the ratio of intima to media (I/M), and the expression of PCNA and PDGF-BB decreased significantly (P < 0.05) in AS-IV groups. Conclusion: AS-IV exerts the inhibitory effects on VSMCs proliferation in a dose- and time-dependent manner. AS-IV significantly inhibits the neointimal hyperplasia in rat carotid artery. It might be partially attributed to the inhibition of proliferation. It may be one of the mechanisms of inhibiting the proliferation of carotid intima in rats induced by balloon injury that AS-IV inhibits the expression of growth factor PDGF-BB.

Isolation and identification of Streptococcus mutans strains with different genotype from clinical samples / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To identify Streptococcus mutans (S. mutans) strains from clinical samples.</p><p><b>METHODS</b>Plaque samples from caries-active and caries-free sites on enamel surfaces were obtained and cultivated for S. mutans isolation. Morphology, biochemistry, automatic microorganism analysis system and polymerase chain reaction using primers homologous to surface protein antigen I/II (spaP), glucosyltransferase B (gtfB) and dextranase (dexA) were used to identify S. mutans. Genotype of isolated S. mutans was determined by arbitrarily primed polymerase chain reaction.</p><p><b>RESULTS</b>Forty-six strains of S. mutans were obtained from the 32 subjects and were identified as S. mutans by biochemistry, automatic microorganism analysis system and polymerase chain reaction. Five identical genotypes were found by arbitrarily primed polymerase chain reaction.</p><p><b>CONCLUSION</b>Forty-one strains of S. mutans with different genotype were obtained from clinical samples.</p>

Pharmacokinetics of diltiazem hydrochloride delay-onset sustained-release pellet capsules in healthy volunteers

The pharmacokinetics (PK) of ordinary tablets and sustained release capsules of diltiazem hydrochloride in human clinical trials had been studied. The PK of diltiazem hydrochloride delay-onset sustained-release pellet capsules, a new dosage form, has not been reported, although it is very important to clinical use. In this paper, we investigated the PK of diltiazem hydrochloride delay-onset sustained-release pellet capsules and the food influence in Chinese healthy volunteers. The PK parameters indicated that the diltiazem hydrochloride delay-onset sustained-release pellet capsules appeared marked characteristics of delayed and controlled release. An opened-label, randomized and parallel clinical trial was conducted in 36 Chinese healthy volunteers with single oral dose (90 mg, 180 mg or 270 mg) and a multiple oral dose (90 mg d-1×6 d) administration. The effect of food on the PK of one single oral dose (360 mg) was investigated in 24 healthy Chinese volunteers. Plasma diltiazem concentration was determined by reversed-phase high-performance liquid chromatography (RP-HPLC) and the main pharmacokinetic parameters were analyzed by PKSolver (Ver 2.0). All clinical studies were conducted in the Clinical Pharmacological Center (No. JDX1999064) of Xiangya Hospital Affiliated Central South University, China. The PK parameters suggested that the new formulation had marked characteristics of delayed and controlled release of diltiazem, and food intake did not alter significantly diltiazem pharmacokinetic parameters.

Embora a farmacocinética (PK) do cloridrato de diltiazem nas formas de comprimidos de liberação imediata e cápsulas de liberação modificada em ensaios clínicos já tenha sido relatada, a pesquisa da PK do cloridrato de diltiazem na forma de cápsulas com peletes de liberação retardada e sustentada ainda é muito importante. Neste trabalho, propusemos avaliar a farmacocinética do cloridrato de diltiazem administrado através desta nova forma farmacêutica em voluntários chineses sadios, assim como a influência da ingestão de alimentos neste perfil farmacocinético. Foi realizado um ensaio clínico aberto, randomizado e paralelo em 36 voluntários, que receberam dose oral única de 90 mg, 180 mg ou 270 mg e dose múltiplas (90 mg/d × 6 d) pela mesma via de administração. Para avaliar o efeito da ingestão de alimentos sobre a PK do diltiazem foi realizada a administração de dose única (360 mg) em 24 voluntários chineses sadios. A concentração plasmática do diltiazem foi determinada por Cromatografia Liquida de Alta Eficiência em fase reversa (CLAE-FR) e os principais parâmetros farmacocinéticos foram analisados através do emprego do software PKSolver (Ver 2.0). O ensaio de farmacocinética clínica foi conduzido na clínica Pharmacological Center (No.JDX1999064) do Hospital de Xiangya, Central South University, China. Os parâmetros PK obtidos indicaram que a nova formulação de cápsulas de liberação retardada e sustentada de cloridrato de diltiazem possue marcantes características de liberação retardada e controlada do fármaco.

Clinical features and differentiations of Parkinson plus syndromes / 中华神经医学杂志

Objective To explore the clinical and imaging features of Parkinson plus syndromes and its differentiation points. Methods Seventy-three patients with idiopathic Parkinson's disease (IPD),68 patients with multiple system atrophy (MSA),10 patients with dementia of Lewy bodies (DLB),15 patients with progressive supranuclear palsy (PSP) and 6 patients with corticobasal degegnration (CBD) were recruited between January,2004 and April,2009 from our hospitals.All patients were given detailed investigation, physical examination, mini-mental status examination and brain CT/MRI examination.Part of patients were performed 18F-FDG PE.Statistical analysis was performed with SPSS 11.0 software. Results Except for postural abnormity, all the other main clinical features differed significantly between each 2 groups (P＜0.05).Brain MRI examination showed that Olivopontocerebellar atrophy was seen in 48 MSA patients (48/59,81.4％),putaminal hypointensities on T2-weighted images were seen in 4 MSA patients (4/59,6.8％) and the "hot cross bun" signal in pons was seen in 30 MSA patients (30/59, 50.8％); hummingbird-like changes were noted in midsagittal view of MRI in 3 PSP patients (3/15, 20.0％); all 6 CBD patients presented asymmetric cortical atrophy, especially in the frontoparietal areas,and 1 also presented putaminal hypointensities on T2-weighted image.Brain 18F-FDG PET indicated that 18F-FDG intake presented different distribution among groups. Conclusion Each atypical Parkinsonian syndrome has its specific clinical features which attribute to rule it out from either IPD or other Parkinson plus syndromes; brain MRI examination and 18F-PET scan can help to diagnose and differentiate Parkinson plus syndromes.

Effects of angiotensin II and its receptor blockers on migration and endothelin-1 expression of rat vascular adventitial fibroblast subpopulations / 药学学报

The study is to investigate the effect of angiotensin II (Ang II) and its receptor blockers on migration and endothelin-1 (ET-1) expression of rat vascular adventitial fibroblast subpopulations. Vascular adventitial fibroblasts were individually expanded by using cloning rings, and the effects of Ang II on the migration of adventitial fibroblast subpopulations were evaluated by Transwell. Fluorescence quantitative-PCR detected the expression of preproET-1 mRNA induced by Ang II, and its receptor antagonists losartan and PD-123319. The concentration of ET-1 was determined by ELISA. It showed that spindle shaped and epithelioid shaped cells were isolated by using cloning rings, named as spindle cells and round cells. RT-PCR showed that fibroblast subpopulations did not have leukocytes, endothelial cells and smooth muscle cells, namely pure cell lines. Compared with respective control cells, two subpopulations had transferring ability. Ang II significantly improved round cells migration in a concentration-dependent manner, and had no obvious influence on spindle cells migration. Ang II (1 x 10(-8) - 1 x 10(-6) mol x L(-1)) significantly increased the expression of preproET-1 mRNA in round cells (P < 0.01), and had no significant effect on the expression of preproET-1 mRNA in spindle cells. Losartan blocked the expression of preproET-1 mRNA induced by Ang II in round cells, and had no significant effect on the expression of preproET-1 mRNA in spindle cells. The effects of Ang II and ET-1 receptor inhibitors on the release of ET-1 were similar to the expression of preproET-1 mRNA. The results indicate that there are two cell subpopulations: round cells and spindle cells in rat vascular adventitial fibroblasts. Ang II significantly improved cells migration, and increased the expression of ET-1 in round cell subpopulation. It suggested that there may be different migratory mechanisms in two cell subpopulations, and the two subpopulations may play a different role in vascular remodeling and reparative process.

A family-based transmission disequilibrium test of polymorphisms of endothelial nitric oxide synthase gene in children with orthostatic intolerance / 中国当代儿科杂志

<p><b>OBJECTIVE</b>Recent studies have found that the variation of G894T on the region of T786C and 7th exon promoted by endothelial nitric oxide synthase (eNOS) gene is associated with cardiovascular disease. This research explored possible correlations between eNOS gene polymorphisms and orthostatic intolerance (OI) in children through linkage disequilibrium analysis between eNOS genes T786C and G894T and OI.</p><p><b>METHODS</b>PCR, Macrorestriction Map and other molecular biotechnology were used to determine the genotypes of eNOS/T786C and G894T in 60 OI probands and their parents. Correlation analysis and transmission disequilibrium test (TDT) between T786C, G894T and OI were performed.</p><p><b>RESULTS</b>There was linkage disequilibrium of eNOS/T786C and G894T gene polymorphisms in the occurrence of childhood OI (P<0.05).</p><p><b>CONCLUSIONS</b>eNOS genes T786C and G894T may be associated with the pathogenesis of OI.</p>

Microgravity-induced cardiovascular deconditioning: mechanisms and countermeasures / 中国应用生理学杂志

It has been demonstrated that individuals exposed to actual or simulated microgravity often experience cardiovascular dysfunctions when returning to Earth. The underlying mechanisms of orthostatic intolerance and countermeasure treatment are still poorly understood. In this paper, the progress in the mechanism of cardiovascular deconditioning from the view of vascular remodeling, increased venous compliance in the lower limbs, cellular proliferation and differentiation, and cell signaling pathway was reviewed. Meanwhile, an overview of the countermeasures including exercise, lower body negative pressure, thigh cuffs, traditional Chinese herb medicine and artificial gravity was presented.

Gene transfection of NGF and/or Noggin affecting the differentiation of bone marrow-derived mesenchymal stem cells / 中华神经医学杂志

Objective To explore the feasibility of exogenous NGF and/or Noggin gene transfecting into the bone marrow-derived mesenchymal stem cells (BMSCs) and observe the differentiation of BMSCs modified by NGF and/or Noggin.Methods BMSCs were isolated from SD rat and purified by adherent method and these cells were identified by their phenotypical properties and their abilities of differentiating into adipocytes.Ad-GFP-NGF and/or Ad-GFP-Noggin were transfected into BMSCs.The protein expressions of NGF and/or Noggin were detected using immunocytochemistry and Western blotting.The differentiations of gene modified BMSCs were observed by immunohistochemistry.Result The cells selected by adherent method had basic phenotypical properties of BMSCs and could differentiate into adipocytes.BMSCs without transfection and those transfected with Ad-GFP expressed low level of NGF without Noggin expression.All gene modified BMSCs could express NGF and/or Noggin.After transfection, BMSCs could differentiate into cells having neuronal morphology and expressing NF (H).The combined transfection group had the highest ratio of NF(H)+ cells among all the groups.Conclusion BMSCs can be isolated and purified from rat bone marrow by adherent method.Ad-GFP-NGF and/or Ad-GFP-Noggin can transfect BMSCs safely and the transfected cells can express those proteins persistently and efficiently.NGF and Noggin can induce BMSCs into neuron-like cells in vitro and when they exist simultaneously,the differentiation is further enhanced.

Comparative study of recurrent colon cancer and recurrent rectal cancer after radical resection / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To explore the difference in tumor biological behaviors and prognosis between recurrent colon cancer and recurrent rectal cancer after radical operation.</p><p><b>METHODS</b>Complete clinical and follow-up data of 132 patients with colorectal cancer developed recurrence,including 36 colon cancers and 96 rectal cancers, after curative resection were retrospectively analyzed and compared with respect of clinical pathological features and prognosis between colon and rectal cancer.</p><p><b>RESULTS</b>Significant differences were found in primary tumor gross type, histological type, tumor differentiation and lymph node metastasis between colon and rectal cancer(P<0.05). Colon cancer recurred earlier than rectal cancer after radical surgery with the median time to recurrence being 14.0 months and 21.5 months, respectively(P=0.028). The difference in multiple sites recurrence was also found between colon(n=16, 44.4%) and rectal cancer(n=65, 67.7%)(P=0.014). The 3-year survival rate of recurrent rectal cancer was better than that of colon cancer (24.8% vs 15.6%, P=0.026).</p><p><b>CONCLUSION</b>There are some differences in tumor biological behaviors between colon and rectal cancer, and the prognosis of rectal cancer with recurrence is better than that of colon cancer.</p>

Effect of intracerebroventricular injection of rAAV-HIF-1α on hippocampal neuronal apoptosis in a rat model of Alzheimer disease / 南方医科大学学报

<p><b>OBJECTIVE</b>To observe the effect of intracerebroventricular injection of rAAV-HIF-1α on hippocampal neuronal apoptosis in a rat model of Alzheimer disease (AD).</p><p><b>METHODS</b>Thirty-two male SD rats (250-300 g) were randomized into 4 groups (n=8), including the normal control group without any treatment, AD model group with right intracerebroventricular injection of 2 µl Aβ25-35 (10 mg/m1), sham-operated group with right intracerebroventricular injection of 2 µl normal saline, and AD+ rAAV-HIF-1α group with right intracerebroventricular injection of 10 µl rAAV-HIF-1a (1×10¹² v.g./m1) one week after Aβ25-35 injection. The rats were sacrificed to detect the expression of HIF-1α and apoptosis of hippocampal neurons 5 weeks after Aβ25-35 or saline injection.</p><p><b>RESULTS</b>Western blotting showed that the expression of HIF-1α was significantly higher in AD+rAAV-HIF-1α group (451.59±34.39) than in normal control group (229.05±41.28) and sham-operated group (216.29±37.08) (P<0.05) without significant difference between the latter two groups. The apoptotic ratio of the hippocampal neurons was significantly higher in AD model group ([19.49±2.59]%) than in normal control group ([5.41±0.75]%) and sham-operated group ([5.28±0.66]%) in (P<0.05), and intracerebroventricular injection of rAAV-HIF-1α resulted in a significant reduction of the apoptotic ratio in the AD rats ([12.07±2.06]%) (P<0.05).</p><p><b>CONCLUSION</b>Intracerebroventricular injection of rAAV-HIF-1α can inhibit hippocampal neuronal apoptosis in the rat model of AD.</p>

Apoptosis in myocyte after repeated + Gz exposures in rats / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To observe the effects of repeated + Gz exposures on the apoptosis of myocyte in rats.</p><p><b>METHODS</b>Twelve male Sprague-Dawley rats were randomly divided into three groups: Control group, + 6Gz group and + 10Gz group. The rats of + Gz groups were exposed to + 6Gz for 3 min, + 10Gz for 3 min respectively, 1 b/d, 1 week. Four control rats were kept at the Earth gravity (1G) in the room with the centrifuge. All animals were anaesthetized and anatomies 1 day after the last exposure. Ventricular myocardium was studied by electron microscopy and terminal deoxynucleotide transferase-mediated dUTP nick end labeling (TUNEL) method.</p><p><b>RESULTS</b>The apoptosis of myocyte in control group and + 6Gz group were scarcely observed by electron microscopy, while heterochromatin concentration and margination were observed in + 10Gz group. The apoptotic index of myocardium increased significantly in + 6Gz and + 10Gz group compared with that of the control group (P < 0.05) and showed the largest value in the + 10Gz group (P < 0.05).</p><p><b>CONCLUSION</b>Repeated + Gz exposures may induce apoptosis in myocyte, and the number of apoptosis in myocyte increases gradually with the increase of G value.</p>

Construction and expression of adeno-associated virus vector harboring human hypoxia inducible factor-1α gene / 第二军医大学学报

Objective: To construct a recombinant adeno-associated virus (AAV) vector harboring human hypoxia inducible factor-1α (HIF-1α) gene and to express it in primarily cultured neurons in the hippocampus. Methods: Human HIF-1α gene was inserted into AAV vector plasmid pSNAV 2.0 to generate a recombinant plasmid pSNAV-HIF-1α, then pSNAV-HIF-1α was transfected into BHK-21 cells and selected by G418. The construction of rAAV-HIF-1α was achieved by infecting G418-resistant BHK-21 cells with recombinant herpes simplex virus. The recombinant rAAV-HIF-1α was confirmed by polymerase chain reaction(PCR). The titer of rAAV-HIF-1α was detected by dot-blot with digoxin labelled cytomegalovirus probe. The purity of rAAV-HIF-1α was detected by SDS-PAGE. The expression of HIF-1α protein was determined by Western blot. Results: PCR demonstrated that human HIF-1α gene was successfully inserted into rAAV-HIF-1α. The Virus titer was 1 × 1012/ml and the virus purity exceeded 98%. Western nblot analysis demonstrated that rAAV-HIF1α infected hippocampal neurons expressed HIF-1α protein. Conclusion: rAAV-HIF-1α has been successfully constructed and it is capable of expressing HIF-1α in cultured hippocampal neurons.

Local hypothermia and optimal temperature for stroke therapy in rats / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Local hypothermia induced by intravascular infusion of cold saline solution effectively reduces brain damage in stroke. We further determined the optimal temperature of local hypothermia in our study.</p><p><b>METHODS</b>Seventy-eight adult male Sprague Dawley rats (260 - 300 g) were randomly divided into 3 groups: group A, ischemia/reperfusion without cold saline infusion (n = 26) (control group); group B, infusion with 20 degrees C saline before reperfusion (n = 26); group C: infusion with 10 degrees C saline before reperfusion (n = 26). In each group, we chose 15 rats for monitoring physical indexes and the temperature of the brain (cortex and striatum) and body (anus), measurement of brain infarction volume, assessment of neurological deficits and the survival rate of reperfusion at 48 hours. Another 8 rats from each group was chosen for examining brain edema, another 3 from each group for histological observation by electron microscopy (EM) and light microscopy (LM) at 48 hours after reperfusion.</p><p><b>RESULTS</b>There was no significant difference among the 3 groups for physical indexes during the examination (F((2, 45)) = 0.577, P = 0.568; F((2, 45)) = 0.42, P = 0.78 for blood pressure and blood gas analysis, respectively). The brain temperature was significantly reduced in the group C compared to the other groups (F((2, 45)) = 37.074, P = 0.000; F((2, 45)) = 32.983, P = 0.000, for cortex and striatum temperature respectively), while the difference in rectal temperature between group A and B or C after reperfusion was not significant (F((2, 45)) = 0.17115, P = 0.637). And the brain infarct volume was significantly reduced in group C (from 40% +/- 10% in group A, 26% +/- 8% in group B, to 12% +/- 6% in group C, F((2, 45)) = 43.465, P = 0.000) with the neurological deficits improving in group C (chi(2) = 27.626, P = 0.000). The survival rate at 48 hours after 10 degrees C and 20 degrees C saline reperfusion was increased by 132.5% and 150%, respectively, as compared to the control group (chi(2) = 10.489, P = 0.005). The extent of the brain edema showed no significant difference (F((2, 21)) = 0.547, P = 0.587) after cold saline infusion compared to the control group. No obvious vascular injury was found by electron or light microscopy in either infusion group.</p><p><b>CONCLUSIONS</b>Regional hypothermia with 10 degrees C cold saline infusion can significantly decrease the infarction volume, improve the neurological deficits, and 10 degrees C seems to be the optimal temperature in inducing a cerebral protection effect during stroke. This procedure could be adopted as a further treatment for acute stroke patients.</p>

Construction of mic2/CD99 gene vector and its transfection in Hodgkin lymphoma L428 cell line / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a eukaryotic expression vector of CD99 gene for transfection into Hodgkin lymphoma L428 cells.</p><p><b>METHODS</b>The full-length cDNA of CD99 gene was amplified from Jurkat cells by RT-PCR and cloned into the pcDNA3.1(+) vector and transfected into L428 cell line using Lipofextamine 2000. The sequence of CD99 mRNA in the transfected cells was confirmed by restriction endonuclease digestion and DNA sequencing, and the expression of CD99 protein was identified using immunocytochemistry.</p><p><b>RESULTS</b>A gene fragment of 558 bp was amplified from the transfected cells and the sequence was verified by DNA sequencing. Immunocytochemistry identified the presence of CD99 expression in the transfected cells.</p><p><b>CONCLUSION</b>A eukaryotic expression vector pcDNA3.1(+)-CD99 is successfully constructed and stably expressed in L428 cell line.</p>

Inhibitory effect of dexamethasone on myeloid differentiation factor 88 and tumor necrosis factor-alpha expressions in mouse peritoneal macrophages / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the inhibitory effect of dexamethasone (DEX) on myeloid differentiation factor 88 (MyD88) and tumor necrosis factor-alpha (TNF-alpha) expression in mouse peritoneal macrophages in innate immune response to Penicillium marneffei (PM).</p><p><b>METHODS</b>Mouse peritoneal macrophages were cultured in the presence of heat-inactivated yeast-phase PM with or without DEX, and the protein and mRNA expressions of MyD88 in the macrophages were detected using Western blotting and real-time PCR, respectively. TNF-alpha in the cell culture supernatant was measured with enzyme-linked immunosorbent assay.</p><p><b>RESULTS</b>DEX suppressed TNF-alpha production by the macrophages co-cultured with PM. The expressions of MyD88 were up-regulated by PM stimulation, whose effect was inhibited by the application of DEX.</p><p><b>CONCLUSION</b>The inhibitory effect of DEX on PM-induced proinflammatory responses of the macrophage is directly associated with the inhibition of MyD88 expression.</p>