Abstract: Objective　To analyze the pathogenic characteristics and epidemiological significance of human plague related strains in Qinghai Province in recent 30 years, so as to provide scientific basis for on-the-spot disposal and prevention and control measures of plague outbreak in Qinghai Province. Methods　A total of 35 strains of Yersinia pestis isolated from 29 typical human plague outbreaks in Qinghai Province from 1980 to 2011 were selected and studied by biochemical fermentation experiments. Virulence factors detection of Fraction 1 antigen (Fra1), virulence antigen (VW), pigmentation (Pgm) and Yersinia pestis Ⅰ (PstⅠ), determinants and genotyping of differential regions (DFRs) were used to study the pathogenic characteristics. At the same time, according to the epidemic situation of human and animal plague in Qinghai Province in recent years, the current situation of plague prevention and control and epidemic characteristics were analyzed. Results　The biotypes of 35 strains of Yersinia pestis were classical, and the biotypes of 29 strains (82.86%) were of Qinghai-Tibet Plateau type, mainly distributed in southern Qinghai and around lake areas, 2 strains (5.71%) belonged to Qilian Mountains type, mainly distributed in Qilian mountains, and 6 genotypes were identified by DFR. Among them, 16 were type 5, 12 were type 8, 2 were type 10, 1 was type 36, 3 were type 30 and 1 was type 1b, the strains of type 5 and 1b were mainly distributed around the lake and the southern foot of Qilian Mountains, while the strains of type 8, 10, 36 and 30 were mainly distributed in the southern part of Qinghai. Conclusions　The pathogen of Yersinia pestis in Qinghai Plateau has complex biochemical types, the epidemic situation among animals is continuous year after year, the situation of prevention and control is serious, the occurrence and prevalence of plague seriously endanger people's health and social development, so it is necessary to do a solid job in the prevention and control of plague to ensure the safety of people's lives.

This study aimed to explore the triterpenic acid components in leaves of Ilex hainanensis. Alkaline water extraction, macroporous resin adsorption, and high performance liquid chromatography were used to separate and purify the triterpenic acid components in leaves of I. hainanensis. The physical and chemical property analysis, MS, NMR spectroscopy, and literature comparison were performed to identify the structures, and a new triterpene acid compound was discovered:(3S, 4R, 5R, 8R, 9R, 10R, 14S, 17S, 18S, 19R)-3,19-dihydroxyursa-12,20(30)-diene-24,28-dioic-acid, and named ilexhainanin F. In addition, according to its structural characteristics, the ~(19)F-NMR Mosher method was further employed to study its absolute configuration. By comparison of the ~(19)F-NMR chemical shifts of Mosher esters, it was determined that the absolute configuration of the 3-position chiral center of the compound was the S configuration.
Chromatography, High Pressure Liquid/methods* ; Ilex/chemistry* ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Leaves/chemistry* ; Triterpenes/analysis*

Accurate methods for identifying pelvic lymph node metastasis (LNM) of prostate cancer (PCa) prior to surgery are still lacking. We aimed to investigate the predictive value of peripheral monocyte count (PMC) for LNM of PCa in this study. Two hundred and ninety-eight patients from three centers were divided into a training set (n = 125) and a validation set (n = 173). In the training set, the independent predictors of LNM were analyzed using univariate and multivariate logistic regression analyses, and the optimal cutoff value was calculated by the receiver operating characteristic (ROC) curve. The sensitivity and specificity of the optimal cutoff were authenticated in the validation cohort. Finally, a nomogram based on the PMC was constructed for predicting LNM. Multivariate analyses of the training cohort demonstrated that clinical T stage, preoperative Gleason score, and PMC were independent risk factors for LNM. The subsequent ROC analysis showed that the optimal cutoff value of PMC for diagnosing LNM was 0.405 × 109 l

This work is to establish the fingerprint of Astragalus membranaceus var. mongholicus by HPLC-ELSD method, and to analyze the simulated wildness degree of A. membranaceus var. mongholicus in the genuine region of Inner Mongolia, Ningxia and Gansu. Compared with wild A. membranaceus var. mongholicus, the quality differences of A. membranaceus var. mongholicus in the genuine region were analyzed by identification of chromatographic peaks and similarity evaluation, cluster analysis(CA), principal components analysis(PCA) and orthogonal partial least squares discriminant analysis(OPLS-DA). HPLC fingerprints of A. membranaceus var. mongholicus in different genuine regions are established. The qualitative analysis of mass spectrometry identified 18 components. The similarity evaluation shows that the similarity of 32 batches of A. membranaceus var. mongholicus samples was 0.688-0.993. Among them, the similarity of samples in Shanxi, Inner Mongolia, Ningxia is 0.688-0.993, 0.835-0.989, 0.934-0.988, respectively and the similarity of samples in Gansu is 0.729-0.876 except No. 25 sample. The results of CA show that the samples of A. membranaceus var. mongholicus can be grouped into four categories according to the production area except the No. 11 and No. 25 samples. The results of PCA indicate that 32 batches of A. membranaceus var. mongholicus samples can be clustered according to quality and origin, and the quality of A. membranaceus var. mongholicus in Inner Mongolia is the closest to the wild breed. The results of OPLS-DA indicate that there are six components that can distinguish the wild and domestic A. membranaceus var. mongholicus, which are malonylastragaloside Ⅰ, astragaloside Ⅰ, calycosin-7-O-β-D-glycoside-6″-O-malonate, calycosin-7-O-β-D-glycoside, formononetin-7-O-β-D-glycoside-6″-O-malonate, and astrapterocarpan-3-O-β-D-glycoside-6″-O-malonate. The established method can be used to analyze differences between A. membranaceus var. mongholicus origin and planting environment, and can provide references for the protection and replacement of wild A. membranaceus var. mongholicus resources, and the cultivation, processing and production of A. membranaceus var. mongholicus.
Astragalus propinquus ; China

In order to explore endophytes diversity and difference in Dendrobium huoshanense,in this paper,the metagenomics method was used to analyze the endophytic bacteria and fungi community of 5 groups include 30 samples in different growth years. The results indicate that 3 540 bacterial OTUs were identified from D. huoshanense,and there are 138 OTUs in 5 groups simultaneously;2 168 fungal OTUs were identified,and 143 OTUs exist in 5 groups simultaneously. The dominate endophytic bacteria community are Sphingomonas sp.,Acinetobacter sp.,Burkholderia sp.,Methylobacterium sp.,Enterococcus sp.,Bacillus sp.,the difference endophytic bacteria community are Oceanobacillusd sp.,Actinomycetospora sp.,Paenibacillus sp.. The dominate endophytic fungi community are Zasmidium sp.,Zymoseptoria sp.,Alternaria sp.,Cladosporium sp.,Fusarium sp.,the difference endophytic fungi community are Cyphellophore sp.,Fusarium sp.. The results of clustering revealed that both the endophytic bacteria and the endophytic fungi,ⅢY2 and ⅢY3 are complete clustered,and ⅡY1 and ⅢY1 are also cluster completely. These enriched the species and resources of endophytic bacteria and fungi in D. huoshanense,and provided a theoretical reference for the reasonable harvest of D. huoshanense.
Ascomycota ; Bacteria ; Dendrobium ; Endophytes ; Fungi ; Fusarium ; Phylogeny

OBJECTIVEThe aim of this study was to investigate the relationship of the mutations of leptin receptor gene exon 4, exon 6, exon9, and exon20 with the tumorigenesis of breast cancer.

METHODSGenomic DNA was extracted from breast cancer tissues of 155 patients, benign lesions of 56 patients and normal tissues and blood samples from 100 health control subjects. The leptin receptor genes were assayed with polymerase chain reaction (PCR) amplification and direct sequence analysis.

RESULTSNucleotide substitutions no mutations were found at exon 4, and nucleotide substitutions occurred at codon 1029 in exon 9, no significant difference among the three groups (P = 0.574). The nucleotide substitutions at codon 668 in exon 6 resulted in Gln223Arg polymorphisms. The occurring frequencies of GG, GA, AA in breast cancer, breast benign lesions tissues and health tissues control group were 70.9% and 17.4%, 12.3%; 80.4%, 14.3% and 5.4%; and 81.0%, 16.0%, and 3.0%, respectively. Alleles of G and A in the three groups were 79.1% and 20.8%, 87.5% and 12.5%, and 89.0% and 11.0%, respectively. Compared the Gln223Arg genotype with the three allele groups, there were significant differences (χ(2) = 16.11, P < 0.005 and χ(2) = 11.41, P < 0.01), respectively. The nucleotide substitutions at codon 3057 in exon 20 resulted in Pro1019Pro polymorphisms. The occurrence frequencies of GG, GA, AA in the breast cancer, benign disease and health control groups were 11.6%, 30.3% and 56.1%; 32.1%, 44.0% and 28.5%; and 32.0%, 45.0% And 23.0%, respectively. Alleles of G and A in the three groups were 26.8% and 73.2%, 51.8% and 48.2%, and 54.5% and 45.5%, respectively. There are significant differences among the three groups (χ(2) = 6.56, P < 0.03 and χ(2) = 5.45, P < 0.05), respectively. Nucleotide substitutions occurred at relatively high frequencies at exon 6 and exon 20 in obese and overweight breast cancer patients compared with those in normal weight breast cancer patients, there were significant differences (P < 0.05 and P < 0.01).

CONCLUSIONSOur findings show that there is no relationship between the variations of leptin receptor gene exon 9 and tumorigenesis of breast cancer. The variation rate of leptin receptor gene exon 6 and exon 20 are significantly increased in the obese and overweight breast cancer patients.

Adenoma ; genetics ; Adult ; Aged ; Breast ; pathology ; Breast Neoplasms ; etiology ; genetics ; Carcinoma ; etiology ; genetics ; Exons ; Female ; Gene Frequency ; Humans ; Hyperplasia ; genetics ; Middle Aged ; Obesity ; genetics ; Point Mutation ; Receptors, Leptin ; genetics

Objective The epidemiological characteristics of congenital heart disease(CHD)among Tibetan children whose age ranged from 4 to 18 at different altitude were investigated in Qinghai province.Methods 32 578 Tibetan children living at 2535 m,3600 m and 4200 m were surveyed with the following 3 steps:prescreened,counterchecked and diagnosed with color Doppler.The entity distribution was then analysed and the age and gender were compared respectively.Resuits 235 CHD cases were identified.The total morbidity was 7.21‰.CHD morbidity was rising with the increase of altitude with 5.45‰ at 2535 m,6.80‰ at 3600 m and 9.79‰ at 4200 m respectively.There were significant static differences between 4200 m and the others with χ2=7.002(P＜0.01)to 2535 m and χ2=5.540(P＜0.05)to 3600 m.However,there was no statistical difference between 2535 m altitude and 3600 m altitude.The morbidity in different age had no statistical difference at 2535 m altitude but statistically increased with the increase of age at 3600 m and 4200 m.The total ratio of 16-18 age was significantly higher than other age periods with χ2=10.79(P＜0.005)to 4-7 age period and with χ2=5.60(P＜0.05)to 8-12 age period.The atrial septal defect(ASD)morbidity rates in three places was 39.1%followed by ventricular septal defect(VSD)with 32.8%and patent duetus arteriosus(PDA)with 24.7%.However,the constitute of CHD was different in different altitudes that VSD with 43.5%at 2535 m.ASD with 42.8%at 3600 m and PDA with 50.8%at 4200 m which was the highest morbidity.Conclusion Morbidity.constitutes and difference in gender and age were related to altitude.

Objective To investigate the clinical features of unexpected sudden death (SUD) clustered in families in Yunnan province.Methods This retrospective study analyzed the clinical features of SUD occurred between July to September 2005 in 7 families in Yunnan province.Results All 16 SUD patients shared common clinical features such as fatigue and repeated syncope and one group of SUD patients (n = 8 from 4 families) presented with the gastric intestinal tract manifestations including nausea, vomiting, abdominal pain and diarrhea with suspected dietary history and abnormal laboratory enzyme findings (GOT/ GPT,CK/CKMB,LDH/LDH1 etc.).In SUD patients without gastric intestinal tract manifestations (n = 8 from 3 families), there were no clear symptoms before death and repeated ventricular tachycardia and ventricular fibrillation were recorded in one survivor.There was no clear evidence for the involvements of hereditary and infectious factors for observed SUD.conclusion The reason for the unexpected sudden death clustered in 7 families in Yunnan remains unclear.Repeated syncope and fatigue served as the common clinical features in the presence or absence of gastric intestinal tract manifestations in all SUD cases.Further studies are needed to clarify the pathology and detailed clinical manifestations of SUD occurred in this area.

The effects of natural immano-active polysaccharide extracts (E) from two mushrooms, Lentinus edodes (LenE) and Tremella fuciformis (TreE), and an herb, Astragalus membranaceus (AstE), on production of the cytokines IFN-γ and IL-2 were investigated in Eimeria tenella infected chickens. A total of 180 broiler chicken were assigned to nine groups: three groups fed with the extracts (LenE, TreE and AstE), three groups fed with extracts and immunized with live oecyst vaccine (LenE + V, TreE + V and AstE + V), a group immunized with oecyst vaccine, and two controls (With or without E. tenella infected). Serum IFN-γ titers and IL-2 production of splenocytes were determined at 7 and 14 days post-infection (p. i. ). The overall mean serum IFN-γ of the groups fed with the extracts and immunized with vaccine was significantly (P ＜ 0.01 ) higher compared with the vaccine group at 7 and 14 days p. i., whereas that of the groups fed with the extracts only was not significantly different from the vaccine group. Of the three extracts, AstE showed the highest IFN-γ titers at 7 days p. i., and the Tree + V group had significantly higher IFN-γ titers compared with both LenE + V and AstE + V groups. The IL-2 production of splenocytes showed a similar pattern to the serum IFN-γ titers. Seven days p. i., the overall mean IL-2 level of the groups fed with extracts and immunized with vaccine was significantly (P ＜ 0.01) higher compared with the group immunized with the oecyst vaccine only, but that of the groups fed with the extracts only were not significantly different from the vaccine group. At 14 days p. i., the overall mean IL-2 production of both the groups fed with the extracts and the groups fed with the extracts and immunized with vaccine was not significantly different from the vaccine group. A significant difference in IL-2 production of splenecytes between different extracts was not found. The use of the mushroom and herb polysaccharide extracts might enhance T-cell immune responses, characterized by IFN-γ and IL-2 secretion, against coccidiosis in chickens, and particularly when they are used in conjunction with vaccine. Thus,these immunoactive herbal polysaccharide extracts may enhance protective immunity against infections.

OBJECTIVETo evaluate the association between serum level of leptin and leptin receptor gene (LEPR) polymorphism and patients with breast cancer.

METHODSLEPR G1n223Arg polymorphism was detected by polymerase chain reaction-restriction fragment length polymorphism in 94 patients with breast cancer and 128 healthy controls. The level of leptin were analyzed by enzyme linked immunosorbent assay.

RESULTSIn univariate regression analyses, we found serum level of leptin and LEPR Gin223Arg genotype polymorphism were significantly higrer than those of the controls (P < 0.05-0.001, respectively). Through multivariable analyses, we found that increased risk estimates for breast cancer were among those with leptin level (OR = 1.53, 95% CI: 1.13-2.07, P = 0.006), LEPR Gin223Arg genotype (OR = 4.87, 95%CI:1.30-18.22, P = 0.019), WHR (OR = 3.68, 95% CI: 1.34-10.11, P = 0.011).

CONCLUSIONResults from this study suggested that LEPR Gln233Agr polymorphism, the elevated WHR and serum level of leptin might be correlated with increased risk of breast cancer.

Breast Neoplasms ; blood ; genetics ; Enzyme-Linked Immunosorbent Assay ; Female ; Genetic Predisposition to Disease ; Humans ; Leptin ; blood ; Lipids ; blood ; Polymorphism, Genetic ; Receptors, Leptin ; genetics ; Risk