Aim To study the expression of caspase-12 mRNA and protein following focal cerebral ischemia-reperfusion in rats,and explore the effect of endoplasmic reticulum pathway on neuronal apoptosis.Methods 60 male Wistar rats were randomly divided into sham-operated group and ischemic group.The middle cerebral artery occlusion(MCAO)models were established by using the intraluminal suture occlusion method,neuronal apoptosis was detected by TUNEL staining,the expression of caspase-12 protein was detected by immunohistochemical staining,the expression of caspase-12 mRNA was detected by RT-PCR method.Results In ischemic group,the number of apoptotic cells and the expression of caspase-12 mRNA and protein were gradually increased following prolonged cerebral reperfusion,reached the peak at 24 h.The number of apoptotic cells and the expression of caspase-12 mRNA and protein in ischemic group were significantly less than those of sham-operated group at all times(P

Diagnosis, Differential ; Humans ; Medicine, Chinese Traditional

Objective: To observe the therapeutic efficacy of music electric stimulation of points in treating anxiety. Methods: By adopting a design of multi-centered randomized controlled trial (RCT), a total of 270 patients with anxiety were randomized into a treatment group and a medication group. The treatment group was intervened by music electric stimulation of points, while the medication group was intervened by oral administration of doxepin. The two groups were evaluated by using Hamilton anxiety rating scale (HAMA) and Chinese revised edition of self-rating anxiety scale (SAS-CR) before and after the intervention. The therapeutic efficacies were also compared. Results: The total effective rate was 93.6% in the treatment group versus 92.3% in the medication group, and the between-group difference was statistically insignificant (P>0.05). After the treatment, the aggregate scores of HAMA and SAS-CR were significantly changed in both groups (both P<0.001), and the inter-group differences were statistically insignificant (P>0.05). Conclusion: Music electric stimulation of points can produce equivalent efficacy in treating anxiety compared to doxepin. Thus, it can be taken as a choice in the treatment of anxiety.

Objective To evaluate the risk of plague occurrence via surveying and analyzing indoor rat density and flea index in natural villages having previous plague experience. Methods During August to September 2007, 30 natural villages experiencing previous plague were selected based on the surveillance data, and then all households were coded with numbers and 20 households in each village were randomly selected via computer. Cages and sticky papers were set in 600 selected households to capture rats and fleas. Rat density, flea prevalence, flea index and median were estimated. Results One hundred thirty-three Rattus flavipectus and 33 Suncus murinus were caught and averaged rat density was 2.8 rats per one hundred cage. nights (166/6000), the median was 5 rats each village. One hundred and one mice infected fleas, flea prevalence on rats was 60.8% (101/166), 296 Xenopsylla cheopis and 48 Leptopsylla segnis were collected. Rat flea index was 2.1 fleas per rat (344/166). A total of 315 dissociated flea was caught, average dissociated flea index was 0.026 fleas per sticky paper (315/11888). The median was 5.5 dissociated fleas per village. Of dissociated fleas, Ctenocephalides felis felis (205) and Xenopsylla cheopis (103) accounted for 97.8% (308/315). The proportion for species of the rat flea and the dissociated flea was different(Fisher test: P < 0.01). The rat flea was significantly associated with the rat density(r = 0.68, P < 0.01), but the dissociated flea was significantly associated with neither the rat density(r = -yield than fried wheat batter(χ2 = 5.59, P < 0.05). Conclusions In these villages having previous plague experience of Lianghe County, Rattusflavipectus was dominant species of indoor rats, Xenopsylla cheopis and Ctenocephalides felis felis were dominant species of rat flea and dissociated flea, respectively. Mengsong, Bangdu, and Tangjiatun village had potential risk of plague emergence.

OBJECTIVETo establish a method for the determination of gossypol in cotton root bark.

METHODSamples were extracted with acetone, and determined on a C18 column with the mobile phase (Acetonitrile-0.2% phosphoric acid, 85:15) and UV-235 nm detector.

RESULTThe recovery rate was 97.6%, RSD 1.59% (n = 5).

CONCLUSIONThis method can be used for the determination of gossypol in cotton root bark and the content of gossypol in three different species has been determined.

Chromatography, High Pressure Liquid ; Gossypium ; chemistry ; Gossypol ; analysis ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the effect of triptolide (TPL) on the renal tissue of diabetic rats and its possible mechanisms.

METHODSSD rats were randomly divided into the normal control group (as the normal group), the diabetic model group (the model group), the low dose TPL treatment group (the low dose TPL group, TPL 0.2 mg/kg by gastrogavage), the high dose TPL treatment group (the high dose TPL group, TPL 0.4 mg/kg by gastrogavage). Equal volume of normal saline was given to rats in the normal group and the model group. Five rats were randomly selected from each group at week 4, 8, and 12 of the experiment to detect body weight, kidney weight, 24 h urinary albumin (24 h UAL), plasma glucose (FBG), total cholesterol (TC), total triglyeride (TG), alanine aminotransferase (ALT), aspartate aminotransferase (AST), white blood cell (WBC), and hemoglobin A1c (HbA1c). The mRNA and protein expression of regulated upon activation normal T-cell expressed and secreted (RANTES) in the renal tissue was assessed by reverse transcription-polymerase chain reaction (RT-PCR) and enzyme linked immunosorbent assay (ELISA). The renal tissue was pathologically stained by HE, PAS, and Masson staining. The glomerular and renal tubular interstitial lesions were observed at each time point. The glomerular sclerosis index (GSI) was observed by PAS staining, and the renal interstitial filrosis index (RIFI) was calcutated.

RESULTSCompared with the same group at week 4, the expression of 24 h UAL, RANTES, GSI, and RIFI at week 12 significantly decreased in two TPL groups (P <0.01). Compared with the same group at week 8, the expression of 24 h UAL, RANTES, GSI, and RIFI at week 12 also significantly decreased in the two TPL groups (P <0. 05, P <0.01). Compared with the normal group, body weight and the kidney weight obviously decreased at week 4, 8, and 12 in the model group (P <0. 01); 24 h UAL, FBG, TG, TC, HbA1c, RANTES, GSI, and RIFI were obviously elevated (P <0.01). Compared with the model group, 24 h UAL, RANTES, GSI, and RIFI also decreased in the two TPL treatment groups (P <0.01). Compared with the low dose TPL group, they were attenuated in the high dose TPL group (P <0. 05, P <0. 01).

CONCLUSIONTPL could not only inhibit the over-expression of RANTES, but also improve the glomerular sclerosis and renal interstitial fibrosis in the renal tissue of diabetic rats.

Animals ; Chemokine CCL5 ; drug effects ; metabolism ; Diabetes Mellitus, Experimental ; drug therapy ; Diabetic Nephropathies ; drug therapy ; Diterpenes ; pharmacology ; Drugs, Chinese Herbal ; metabolism ; Epoxy Compounds ; pharmacology ; Glycated Hemoglobin A ; metabolism ; Immunosuppressive Agents ; pharmacology ; Kidney ; drug effects ; Kidney Diseases ; drug therapy ; Kidney Glomerulus ; metabolism ; Kidney Tubules ; metabolism ; Phenanthrenes ; pharmacology ; RNA, Messenger ; genetics ; Rats

Objective To construct cDNA entry library and cDNA expression library of Armillifer agkistrodontis (A.) nymphs and make a preliminary immunoscreening for the cDNA expression library.Methods The nymphs were collected from the Kunming mice infected experimentally with A.agkistrodontis eggs and the total RNA were extracted from the nymphs using TRIzol Reagent.After purifying the mRNA,the synthesized cDNAs were cloned into the donor vector pDONR222 by BP reaction of Gateway technology and the recombinants were transformed into the DH10B cells by electroporation,the cDNA entry library was obtained.Next,the expression vector pDEST17 was ligated with entry clones by LR reaction,and the recombinants were transformed into the BL21 (DE3) cells.Hence,the cDNA expression library was constructed.Then,the expression library was immunoscreened with the mixed sera of mice infected with A.agkistrodontis,and the insertions of positive clones were sequenced.After that,the open reading frame(ORF) of positive slone sequence,the homology of the screened genes and their encoded proteins were analyzed by Finder and BLAST (basic local alignment search tool) program of National Center of Biotechnology Information(NCBI),and the discovered new genes were submitted into the GenBank.Besides,the physico-chemical properties,secondary structure and B cell epitopes of encoded proteins were also analyzed by bioinformatics software.Results The average titer and total clones of the cDNA entry library were 1.45 × 105 CFU/ml(colony-forming unit,CFU) and 1.74 × 106 CFU,respectively,and the range of fragment length of the inserted cDNA was between 0.2-4.0 kb,with an average of 1.4 kb.The total clones of cDNA expression library were 1.00 × 105 CFU,and the fragment length of the inserted cDNA was between 0.3-2.2 kb,with an average of 1.0 kb.Five positive clones,coded S1,S5,A1,D1 and F1,respectively,were obtained through preliminary immunoscreening.The sequence and homology of the five positive clones were sequenced and analyzed by BLAST program.No significant similarities were found in pentastomida species,which meant that they were all novel genes of A.agkistrodontis.The gene sequences were submitted to GenBank,with the accession number from JQ180451 to JQ180455.Also,results obtained by bioinformatics software showed that the predictive encoding proteins were all potential to be valuable recombinant diagnostic antigens.Conclusions The cDNA library of A.agkistrodontis nymphs is successfully constructed,and five new genes of A.agkistrodontis are discovered.The establishment of cDNA library and the discovery of the new genes will lay a foundation for further studying the gene functions and screening the immunodiagnostic antigens.

Objective: To summarize the influence patterns of related time factors on acupuncture effectiveness in different disease model rats/mice, and to provide reference for acupuncture clinical practice. Methods: Retrieved the relevant literatures on time-effect experimental studies of acupuncture in rats/mice in the recent 10 years. The correlations between the key time factors (such as different intervention timings of acupuncture, acupuncture moments, operation durations, needle-retaining times, intervals and treatment courses) and the acupuncture effect were analyzed and summarized. Results: From the mainstream perspective of quantification, the earlier the acupuncture intervention, the better. The proper time to implement acupuncture varied depending on disease models and points. The best operation time varied widely between different diseases. The most frequently needle-retaining time was 20-30 min. The frequency of acupuncture was usually 1 time/day; the length of the treatment course was determined according to practitioners' experience. Conclusion: Throughout the time-effect studies of acupuncture intervention in experimental rats/mice, conclusions are inconsistent, especially the lack of quantitative research on acupuncture operation time, acupuncture frequency, acupuncture treatment duration, and optimal stimulation amount. Future research should explore and determine the best time-quantity parameters that affect the effectiveness of acupuncture intervention, which is the key and goal of the acupuncture time-effect research. Independent intervention-time experiments throughout the entire course of a single disease (dominant disease) need to be done to guide clinical and disciplinary development.

96%).There are 14 nucleotide mutations between yak and cattle and 3 of that were predicted to alter the protein sequence,both of which occurred in the domain III.Construction of molecular phylogenetic tree shows:The cluster results coincided with the traditional taxonomy.

Objective:To investigate the expression of wild type estrogen receptor(wER)and the ex-on-5 deleted ER(variant ER,vER)in human hepatocellular carcinoma(HCC)samples,and thereafteranalyze the possibility of HCC treatment by endocrine therapy.Methods:The mRNA expressions of wERand vER were analysed from 28 cases of HCC by RT-PCR.The expression of ER at the protein level wasdetected by immunohistochemistry(IHC).Results:IHC results showed that 39.3% of the HCC speci-mens expressed ER.The mRNA of wER was detected in 89.3%(25/28)of the HCC specimens whilethat of vER was detected in 96.4%(27/28).Twenty four out of 28 HCC cases(85.7%)expressedboth wER and vER.One out of 28 patients(3.5%)expressed only wER whereas 3 patients out of 28(10.7%)expressed vER only.Conclusion:Ninety six percent(27/28)of the HCC patients expressedvER,which suggests that the expression of vER is an important event in the development of HCC.