Humans ; Polyomavirus ; classification ; Polyomavirus Infections ; virology

Objective To compare the effect of 4 different cold and hot properties of traditional Chinese medicine (TCM) on body temperature and related factors of yeast induced fever rats,and discuss the thermoregulatory mechanism of cold and hot properties of TCM.Methods 108 male SD rats were randomly divided into a normal group,a yeast-induced group,a R.palmatum treated group,a C.chinensis treated group,a Euodia ruticarpa treated group,and a Alpinia officinarum Hance treated group,with 18 rats in each group.Pyrexia model was induced by injecting yeast suspension subcutaneously on rat.At the 4h,8h and 12h after injection of yeast,the rats were sacrificed,and the blood and hypothalamus were collected.The levels of prostaglandin E2 (PGE2),cyclic adenosine 3',5'-monophosphate (cAMP) and arginine vasopressin (AVP) in hypothalamus and plasma were detected by ELISA assay.Results At the 4h after injection of yeast,the temperature of rats in the model group began to rise,and it reached the peak at 8h,while RheumpalmatumL and Coptis chinensis could significantly reduce the body temperature of yeast-induced rat (P＜ 0.01 or P＜ 0.05).At 8h,the levels of PGE2 and cAMP in hypothalamus increased significantly [respectively (31.55 ± 9.88) pg/mg and (0.17±0.03) pmol/mg] compared with the normal group,while the level of AVP (0.14±0.02) pmol/ml in plasma reduced (P＜0.05).Compared with model group,at 8h RheumpalmatumL and Coptis chinensis could significantly lowered PGE2 [respectively (113.65± 18.60) pg/mg and (127.72 ± 15.75) pg/mg,P＜ 0.01 or P＜0.05],and cAMP [respectively (0.69±0.08) pmol/mg and (0.74±0.10) pmol/mg,P＜0.05] in hypothalamus,and increased AVP levels [respectively (1.08 ± 0.12) pmol/ml and (0.91 ±0.01) pmol/ml,P＜0.05 or P＜0.01] in plasma.Euodia ruticarpa and Alpinia officinarum had no significant effect on both body temperature and the levels of inflammatory factors.Conclusion The two cold property traditional Chinese medicines,R.palmatum and C.chinensis,could significantly reduced the body temperature of yeast-induced rats,which may be related to its effective regulation on levels of PGE2 and cAMP in hypothalamus and AVP in plasma,however,the two hot property traditional Chinese medicine,Euodia ruticarpa and Alpinia officinarum Hance,had no related effects.

BackgroundRecent studies showed that diabetic retinal neuropathy is an earlier and more dangerous complication and neurotrophin has a protective effect on retina.ObjectiveThe present study was to observe the changes of brain derived neurotrophic factor (BDNF),its receptor TrkB,signal pathway protein phosphatized extracellular signal-regulated protein kinase1/2 (p-ERK1/2) and c-fos in the retina after injection of BDNF into the vitreous in STZ induced Wistar diabetic rats.MethodsWistar rats aged 9 weeks-old were randomly divided into BDNF injection group,diabetes mellitus (DM) control group and normal control group and 20 rats for each group.STZ was intraperitoneally injected in the rats of BDNF injection group and DM control group to create the experimental DM.BDNF was intravitreously injected in the rats of BDNF group 2 weeks after administration of STZ in three-day interval for 5 times,and BSS containing O.1％ bovine serum albumin (BSA) was used at the same way in the DM control group and normal control group.The retina was isolated for hybridization in situ for BDNF,and TrkB,p-ERK1/2 and c-fos.Levels in retina were detected using sandwich method ELISA.ResultsThe number of BDNF positive cells and the gray scale were lower obviously in the rat retina of DM control group than those of BDNF injection group and normal control group,showing significant differences among the 3 groups ( F =102.36,92.55 ;P＜0.05 ).ELISA assay showed that TrkB,p-ERK1/2 and c-fos values in retina were statistically significantly different among the 3 groups ( F =92.54,95.46,94.84,P＜0.05 ).The TrkB level in retina was statistically reduced,but the p-ERK1/2 and c-fos levels in retina were increased statistically in DM control group compared with BDNF injection group and normal control group( P＜0.05 ).No statistical difference was found in TrkB,p-ERK1/2 and c-fos values between the BDNF injection group and normal control group(P＞0.05).ConclusionsThe injection of BDNF into the vitreous cavity can protect retina from downregulating BDNF and TrkB levels and up-regulating the p-ERK1/2 and c-fos protein levels in the early stage of DM.

Objective:To explore the genetic polymorphism of cytochrome P450 enzyme 17(CYP17) and its relationship with the pathogenesis ofpolycystic ovary syndrome(PCOS).Methods:A total of 260 patients with PCOS of uyhan and han nationality in Xinjiang,who were admitted to Chinese Medicine Research Institute of Xinjiang Uygur Autonomous Region from January 2015 to February 2017,were chosen as PCOS group;the other 237 healthy fertile women,who were examined in the outpatient department during the same period,were chosen as control group.The CYP17 gene polymorphism in the two groups of subjects was detected by the restriction fragment length polymorphism polymerase chain reaction (PCR-RFLP) technique;the distribution of alleles and gene frequencies was compared between the two groups;combining with its clinical data,the relationship between CYP17 gene polymorphism and the pathogenesis of PCOS was analyzed.Results:Body Mass Index(BMI) and Follicle-stimulating hormone(FSH) levels in the PCOS group were lower than those in the control group;Luteotropic hormone(LH),Testosterone(TES) and LH/FSH levels in the PCOS group were higher than those in the control group,and the differences were statistically significant (P＜0.05).The frequencies of A1A1,A1A2,and A2A2 of CYP17 gene in the PCOS group were 34.62％,41.92％,23.46％,respectively,compared with 34.18％,43.88％,21.94％ in the control group,the difference was not statistically significant (P＞0.05).The allele frequency of A1 and A2 in the PCOS group were 55.58％,and 44.42％ respectively,compared with 56.12％ and 43.88％ in the control grouP,the difference was not statistically significant (P＞0.05).There was not statistical significance in FSH,LH and LH/FSH levels of different CYP17 genotypes in the PCOS group (P＞0.05).There was statistical significance in BMI level A2A2＞A1A2＞A1A1 and TES level A2A2＜A1A1＜A1A2 in the PCOS group (all P＜0.05).The BMI,FSH,LH,TES and LH/FSH levels of different CYP17 genotypes in the control group were not statistically significant (P＞0.05).Conclusion:CYP17 gene-34bp T/C nucleotide polymorphisms is a common base replacement in the population,which is not significantly related to the pathogenesis of PCOS.

The marine biological source of mineral drugs recorded in Chinese Pharmacopoeia (2010 version) mainly including pearl, nacre, clam shell, common oyster shell, ark shell, cuttle bone, and sea-ear shell are widely used in clinical. Calcium carbonate and a small amount of protein are the main components in this type of drugs. In this paper, a systematical and comparable study were carried out by determination of calcium carbonate by EDTA titration method, the crystal of calcium carbonate by X-Ray powder diffraction and the total amino acids (TAAs) of the hydrolyzed samples by ultraviolet spectrophotometry method. As a result, the crystal structure is calcite for common oyster shell, mixture of calcite and aragonite for nacre and sea-ear shell, aragonite for the other drugs. The content of calcium carbonate ranged from 86% to 96%. Cuttle bone has the highest amount of TAAs among the seven drugs which reached 1.7% while clam shell has the lowest content of 0.16% on average. In conclusion, an effective method was developed for the quality control of marine mineral drugs by comprehensive analysis of calcium carbonate and TAAs in the seven marine mineral drugs.
Amino Acids ; analysis ; chemistry ; Animal Shells ; chemistry ; Animals ; Calcium Carbonate ; analysis ; chemistry ; Crystallization ; Edetic Acid ; chemistry ; Mollusca ; chemistry ; classification ; Pharmaceutical Preparations ; analysis ; chemistry ; standards ; Quality Control ; Reproducibility of Results ; Seawater ; Species Specificity ; Spectrophotometry, Ultraviolet ; X-Ray Diffraction

Objective To observe effects of different extracts of Jianpi Huogu Formula (JPHGF) on proliferation and differentiation of bone marrow mesenchymal stem cell (BMSCs). Methods Whole bone marrow adherent was used to screen, culture, and isolate BMSCs. Extracts from different parts (water, chloroform, ethyl acetate and n-butanol parts) of JPHGF were administrated for a certain time. MTS was used to detent cell proliferation;ALP staining was used to detect ALP activity;ARS staining was used to detect the formation of calcium nodules;oil red O staining was used to detect fat cell formation. Results Extracts from different parts of JPHGF could promote cell proliferation of BMSCs in different levels, followed by its strength in water, chloroform, ethyl acetate, and n-butanol parts;ALP staining results showed that the intensity of ALP expression of the order is water, acetic acid ethyl, chloroform and n-butanol parts;in promoting the formation of calcium nodules, ARS staining results showed that its intensity were water, chloroform, ethyl acetate, and n-butanol parts;oil red O staining results showed that inhibition intensity of fat cells interaction strength was formed from ethyl acetate, water, chloroform to n-butanol parts. Conclusion Extracts from different parts of JPHGF have different effects on BMSCs proliferation and differentiation. Water extraction has the strongest osteogenic differentiation and proliferation, and ethyl acetate has the best effect on the inhibition of cell formation.

Objective To construct a recombinant bacterial vaccine which can express specific 10-23 deoxyribozyme(DZ) in macrophage, identify the intracellular production of specific 10-23DZ and detect the activity of this recombinant bacterial vaccine on inhibiting the expression of TACO gene in macrophage.Methods The pSDE02 was obtained by inserting the replicon of Mycobacterium into pSDE01, a plasmid which can express 10-23DZ in eukaryotic cells. The expression sequence of DZ1, a 10-23DZ targeting the TACO mRNA of macrophage designed in our previous study was synthesized and inserted into pSDE02. The resulted plasmid was named pDZM01. pDZM01 was then transferred into Mycobacterium smegmatis by electroperation. The recombinant M. smegmatis, named rMs-DZ1 was screened on low-salt LB medium containing Zeocin and identified by Colony PCR. The targeted delivery property of recombinant M. smegmatis was observed by Ziehl-Heelson stain and GFP expression observation via fluorescence microscope. rMs-DZ1 was used to infect RAW264.7 cells and the expression of DZ1 in macrophage was identified by dot-blot assay. At 24 h and 48 h after infection, total RNA and proteins were extracted and the TACO mRNA and protein expression level was assayed by RT-PCR and western-blot respectively. Results Restrictive analysis and sequencing data showed that the Mycobacterium-eukaryotic cell shuttle plasmid pSDE02 and pDZM01 was successfully constructed. rMs-DZ1 was confirmed by colony PCR. When engulfed by macrophage, rMs-DZ1 would express DZ1 in RAW264.7 cells and inhibit the expression of taco gene. When compared to uninfected macrophage, rMs-DZ1 significantly reduced the taco mRNA by 67.90% and 57.14% and down-regulated the expression of TACO protein by 53.85% and 68.92% at 24 h and 48 h respectively. Conclusion A recombinant M. smegmatis vaccine was successfully constructed which could generate specific 10-23DZ in macrophage and inhibit the expression of target gene of interest. To our knowledge, this is the first bacterial vector which can express intracellularly 10-23DRz in targeted manner. This study may further prompt the feasibility of using 10-23 DNAzyme to achieve effective and targeted gene silence.

Background There are a lot of studies about the carrier of corneal endothelial transplantation,but the best carrier has not been defined.Objective This study was to investigate the biocompatibility of chitosan carrier with rabbit corneal endothelium in vivo.Methods Fresh eye-balls were obtained from 10 New Zealand white rabbits.Rabbit corneal endothelial cells (CECs) were isolated and cultured on chitosan carrier in vitro.The morphology and density of rabbits CECs were observed every day,and the expressions of fibronectin (FN),collagen-1 (Coil-I) and Zonula occludens 1 (ZO-1) were detected by immunoinfluorescence.The morphology and ultrastructure of CECs were observed under the scanning and transmission electron microscope.Chitosan carrier with CECs was implanted into the anterior chamber of the left eyes in ten healthy New Zealand white rabbits,and only paracentesis of anterior chamber was performed in the right eyes as controls.The inflammation of ocular anterior segment was examined under the slit lamp microscope,and corneal thickness was measured 1 week,4 and 8 weeks after operation.Corneal endothelium cell density and morphology were examined under the corneal endothelial microscope at postoperative 2 weeks.Corneal samples were collected for the regular histopathological examination to observe the inflammatory reaction at postoperative 1 month and 3 months.Paired t test was used for statistical analyses between the control group (left eyes) and the experimental group (right eyes).The use and care of the animals followed the Statement of ARVO.Results CECs formed an intact monolayer of cells with the uniform shape and size on the chitosan membrane after incubated for 5 days.The cells reached confluence of 90％ 7 days after cultured with the 40％ hexagon cells.Under the scanning electron nicroscope,rabbit CECs showed the round or polygon in the shape with the microvillus on the cell surface.The cells connected closely by desmosome.The processes,pseudopodiums and microvillus on the cellular surface,vacuole in the cytoplasm,expanded endoplasmic reticulum with ribosome and abundant chromatin were exhibited under the transmission electron microscope.The immunofluorescence examination revealed the positive expressions of FN,Coll-Ⅰ and ZO-1 in the CECs on the chitosan carrier.In the in vivo experiment,the exudation in the anterior chamber and corneal edema were seen under the slit lamp microscope 3 days after implantation of chitosan carrier with CECs.However,the inflammation was gone 14 days after operation.The differences of the corneal thickness were no significant between the experimental group and the control group 1 week and 4,8 weeks after operation (t =1.377,P=0.265;t =1.795,P=0.165 ; t =0.390,P =0.760).In addition,no significant differences were found in the CECs density and the hexagon cells rate between the two groups(P =0.365,0.062).The histopathological examination showed that the inflammatory cells around the chitosan membrane were disappeared 3 months after operation and showed a good corneal structure.Conclusions Chitosan carrier has a good biocompatibility with rabbit CECs and anterior chamber,and it may be a potentially good carrier for CECs transplantation.

AIMTo explore the role of ligustrazin on dynamic changes of lipid peroxidation in hepatic ischemia/reperfusion injury (HIRI) and its mechanism.

METHODSThe HIRI model was used. Twenty rabbits were randomly divided into control group (n = 10) and ligustrazin group (n = 10). The xanthine oxidase (XO) activity, superoxide dismutase (SOD) activity,malondialdehyde (MDA) content and glutamic pyruvic transaminase (GPT) activity in plasma were observed before ischemia and at ischemia 25 min, reperfusion 25 min, reperfusion 60 min and reperfusion 120 min.

RESULTSThe XO activity, SOD activity, MDA content and GPT activity of ligustrazin group, as compared with control group, showed significant differences (P < 0.05 or P < 0.01) at total time points of reperfusion.

CONCLUSIONLigustrazin has notable anti-lipid peroxidation effect on HIRI, which is due to its inhibiting the generation of oxygen free radicals and its strengthening scavenging of oxygen free radicals.

Alanine Transaminase ; metabolism ; Animals ; Female ; Lipid Peroxidation ; drug effects ; Liver ; drug effects ; metabolism ; Male ; Malondialdehyde ; blood ; Pyrazines ; pharmacology ; Rabbits ; Reperfusion Injury ; metabolism ; Superoxide Dismutase ; metabolism ; Xanthine Oxidase ; metabolism