基于数据挖掘“疫”病方剂用药规律对新型冠状病毒肺炎防治的用药启发

目的:基于数据挖掘技术探究中药复方治疗"疫"病的用药规律,为新型冠状病毒肺炎的防治提供中药配伍的用药启发。方法:以《中国方剂数据库》为检索库,检索治疗"疫"病的中药方剂,筛选中药复方治疗疫证方剂数据库,并采用Excel 2013,SPSS 21.0及SPSS Modeler 18.0等软件进行频数及频率分析、主成分分析、聚类分析、关联分析、相关性分析等,进而对中药复方用药规律进行定量、定性分析。结果:共收集治疗"疫"病方剂177首,对古代方剂功效主治分析发现,"疫"病方剂主要用于头痛、发热、咳嗽、腹泻等临床情况与新型冠状病毒感染所致的肺炎症状相似,具有参考意义。其中,复方多见于汤、散、丸、丹等剂型;高频中药54味,甘草、半夏、陈皮、白芍、藿香等中药较为常用;且新型冠状病毒肺炎诊疗各阶段推荐处方中高频药物的数量分布频率较高;177首方剂含有药物数量集中在4味~11味之间,而方剂中含有高频率药物集中在3味~8味之间;解表药、清热药、补虚药最常选用;四气以温、微寒、寒为主,五味以辛、苦、甘为主,归经以归肺经、脾经、胃经、肝经为主;主成分分析提取了18个公因子,累积贡献率达69.04%,结果显示以厚朴、藿香、羌活、陈皮、白芷等药为主;聚类分析结果显示分为5类;关联分析共得到34条关联规则,药对以厚朴→甘草、麝香→冰片、白芍→半夏等为常用,三味药组以厚朴+陈皮→甘草,厚朴+苍术→甘草,厚朴+藿香→甘草等为常用。相关分析有显著意义的共410个药对,分别为连翘+天花粉(0.75)、雄黄+朱砂(0.72)、玄参+天花粉(0.71)正相关最显著。结论:古代治疗"疫"病多用温、寒,辛、苦、甘味药,从肺脾胃入手,以祛邪扶正为主要治则,多采用解表散寒、清热解毒结合温阳益气,滋阴养血、化痰止咳、行气活血等方法选取相应中药组方治疗。

论“疫情”的新义

"疫情"的意义本是"疫病的发生和发展情况",但在时下的语言交际中已经产生了"流行性传染病"的新义。"疫情"在旧义和新义上,要求意义不同的词语与之搭配,"疫情"在新义上和"瘟疫""疫病"的搭配范围几乎重合,可以视为后者的同义词。"疫情"中的"情"意义很空泛,发话人为了委婉的表达,直接用"疫情"中的"疫"指代"瘟疫","疫情"就产生了新义。

类风湿性关节炎膝关节病变的X线影像学研究

目的 :分析类风湿性关节炎(RA)膝关节病变的X线影像学特征,以提高诊断正确率。方法 :收集2015年2月至2017年2月收治的RA患者52例和骨关节炎患者45例,进行X线摄片,分析两组膝关节病变的影像学特征。结果 :RA组中,关节间隙呈均匀性狭窄47例(90.4%),髌上囊肿胀41例(78.8%),关节面毛糙、关节面不同程度侵蚀破坏22例,关节周缘轻度骨质增生43例,明显重度增生、骨赘形成5例,未见明显骨质增生4例,52例均有不同程度骨质疏松。OA组中,关节间隙呈均匀性狭窄5例(11.1%),髌上囊肿胀16例(35.6%),关节面下小囊性改变7例,45例有不同程度骨质增生、骨赘形成改变。两组关节间隙均匀狭窄及髌上囊肿胀改变发生率差异有统计学意义(P<0.05)。结论 :RA膝关节病变的X线影像学表现具有一定特征性,尤其是关节间隙均匀性狭窄、髌上囊肿胀及关节面侵蚀的改变在鉴别RA和OA的诊断中有重要价值。

Two-sample Mendelian randomization analysis of causal relationship between eczema and autoimmune diseases

Objective:The causal relationship between eczema and autoimmune diseases has not been previously reported.This study aims to evaluate the causal relationship between eczema and autoimmune diseases.Methods:The two‐sample Mendelian randomization(MR)method was used to assess the causal effect of eczema on autoimmune diseases.Summary data from the Genome-Wide Association Study Catalog(GWAS)were obtained from the Integrative Epidemiology Unit(IEU)database.For eczema and autoimmune diseases,genetic instrument variants(GIVs)were identified according to the significant difference(P<5×10−8).Causal effect estimates were generated using the inverse‐variance weighted(IVW)method.MR Egger,maximum likelihood,MR-PRESSO,and MR-RAPS methods were used for alternative analyses.Sensitivity tests,including heterogeneity,horizontal pleiotropy,and leave-one-out analyses,were performed.Finally,reverse causality was assessed.Results:Genetic susceptibility to eczema was associated with an increased risk of Crohn’s disease(OR=1.444,95%CI 1.199 to 1.738,P<0.001)and ulcerative colitis(OR=1.002,95%CI 1.001 to 1.003,P=0.002).However,no causal relationship was found for the other 6 autoimmune diseases,including systemic lupus erythematosus(SLE)(OR=0.932,P=0.401),bullous pemphigoid(BP)(OR=1.191,P=0.642),vitiligo(OR=1.000,P=0.327),multiple sclerosis(MS)(OR=1.000,P=0.965),ankylosing spondylitis(AS)(OR=1.001,P=0.121),rheumatoid arthritis(RA)(OR=1.000,P=0.460).Additionally,no reverse causal relationship was found between autoimmune diseases and eczema.Conclusion:Eczema is associated with an increased risk of Crohn’s disease and ulcerative colitis.No causal relationship is found between eczema and SLE,MS,AS,RA,BP,or vitiligo.

Cloning of Potato POTHR-1 Gene and Its Expression in Response to Infection by Phytophthora infestans and Other Abiotic Stimuli

A complete cDNA of potato Phytophthora infestans-induced hypersensitive response-related protein gene (POTHR-1) was cloned using rapid amplification of cDNA ends (RACE) strategy according to a fragment sequence which we had cloned using suppression subtractive hybridization (SSH) technique. The potato POTHR-1 gene encodes a protein of 225 amino acids, which shares 81% identity with tobacco hin1 gene-enoded protein (harpin-induced protein). Southern blot revealed that there are two to three copies of POTHR-1 in potato genome. The POTHR-1 gene expression in potato leaves showed that its transcripts accumulated remarkably in leaves after 36 h inoculation with P. infestans. Mechanical wounding and jasmonic acid (JA) could induce the POTHR-1 gene expression and osmotic stress just induce a slight accumulation of POTHR-1 gene mRNA, while salicylic acid (SA) had no detectable function on the induction accumulation of POTHR-1 gene transcripts. The potato POTHR-1 gene may preferentially associate with hypersensitive response (HR) or biotic cell death during interaction between host and pathogen.

Expression and Identification of the Type-specific Antigen of FMDV of Serotype C

[Objective]The aim was to provide a theoretical basis for preparation of polyclonal antibodies and monoclonal antibody that of type specificity, as well as Foot-and-mouth Disease Virus （FMDV） typing.[Method] The recombinant plasmid pGEM-CP1 that contained VPI gene of FMDV of serotype C was used as template for the VP1 and its C terminus coding fragments of FMDV of serotypes C amplification. The coding fragments of VP1 and its C terminus were respectively cloned into prokaryotic expression vector for prokaryotic expression and the reactionogenicity was detected. The purified fusion protein of FMDV VP1 and its C terminus of serotype C were used to construct the indirect ELISA method to detect positive sera of four serotypes A, O, C and Asia 1 of guinea pig and determine the cross reactivity of FMDV antibody of VP1 and its C terminus of serotype C with other three serotypes. [ Result] The recombinant prokaryotic expression plasmids of PPRO-CVP1 and pPRO-CVPlc were constructed, FMDV VP1 and its C terminus of serotype C were expressed in high level, and the molecular weight of target proteins was 33 kD and 20 kD respectively. Western blot result showed that the fusion protein of VP1 and its C terminus could react with the positive sera of guinea pig of the same serotype. ELISA results revealed that VP1 and its C terminus of serotype C are type-specific and no cross-reactivities were shown between guinea pig positive sera of FMDV of serotype C with the other three serotypes, and the C terminus showed better type-specificity. [ Conclusion] FMDV specific antigen of serotype C was obtained.

Expression,Purification and Activity Detection of VP1 of A-type FMDV

[Objective] The research aimed to induce the expression of FMDV structural protein VP1 in E.coli and purify the protein,then detect the activity.[Method] The fragment coding VP1 was amplified by PCR and doubly digested with BamH Ⅰ and XhoⅠ,then cloned into expression vector pGEX-4T-1 and pPROExHTb respectively to get recombinant plasmid pGEX-4T-1-VP1 and pPROExHTb-VP1.The recombinant plasmid pGEX-4T-1-VP1 and pPROExHTb-VP1 was transformed into E.coli BL21(DE3)and induced by IPTG,fusion protein was identifie...

Establishment of Monoclonal Antibody Competitive ELISA Using Monoclonal Antibody Against VP1 Protein of Asia 1 Type Foot-and-Mouth Disease Virus

Using the purified VP1 protein of Asia 1 type foot-and-mouth disease virus as the antigen, the purified monoclonal antibody was labeled by the sodium periodate method and the monoclonal antibody competitive ELISA was established in this study. Ten positive porcine foot-and-mouth disease serums and more than two hundreds negative serum were tested, and the results were the same as the background of samples. The sensitivity test and replicate test indicated that this method was stable and sensitive, which was suitable for monitoring Asia 1 type porcine foot-and-mouth disease virus antibody.

Immunologic pathogenesis of multiple sclerosis

Multiple sclerosis （MS） is an autoimmune disease. The etiology and pathogenesis of MS remain unclear. At present, there are substantial evidences to support the hypothesis that genetics plays a crucial role. The people who have genetic predisposing genes easily develop immune-mediated disorder, probably in conjunction with environmental factors. The aim of this review is to describe recent observations regarding the immunologic pathogenesis of MS.

Construction and Identification of a Goat Pox Virus Transfer Vector to Express Peste des Petits Ruminants H gene

[Objective] This study was to develop a live vector vaccine of goat pox virus of Peste des petits ruminants(PPR). [Method] Using PCR amplification technique, PPR H gene was obtained, then ligated into pGEM-T easy vector; the recombinants were digested by Nhe Ⅰ and Hind Ⅲ, and ligated into pEGFP-N1-P7.5, yielding the recombinant vector pEGFP-N1-P7.5-H; next the expression cassette EGFP-N1-P7.5-H was first released from recombinant vector pEGFP-N1-P7.5-H by double digestion of Hind Ⅲ and Nhe Ⅰ and ligated into pUC119-TK that was digested by Kpn Ⅰ, yielding the transfer vector pUC119-TK-EGFP-P7.5-H. [Result] Identification and double enzyme digestion showed that the transfer vector pUC119-TK-EGFP-P7.5-H was correctly constructed. From the transfer vector transfected BHK-21 cells which infected GTPV AV41, specific fluorescence was observed at 48th h of transfection. [Conclusion] The construction of goat poxvirus live vector laid a foundation for the live vector vaccine of PPR vaccine.

Study on Inactivated Vaccine in Oil Emulsion Against Newcastle Disease and Fowl Cholera IV——Field Test of Vaccine

[ Objective] The aim of this study was to evaluate the clinical effect of the inactivated vaccine in oil emulsion against Newcastle disease and Fowl cholera, and provide conditions for combined prevention and control of Newcastle disease and Fowl cholera. [ Method] The mixture of avian pasteurella multocida （type A） virulent strain 1502 and Newcastle disease virus attenuated strain La Sota was prepared into five batches of the inactivated vaccine in oil emulsion to use in the field test for assessing its safety and effects on immune protection of chicken, duck and goose. [ Result] The field safety test showed that there was no adverse reaction in the vaccinated chickens, ducks and geese. The field test of immune effect for chickens suggested that the titers of hemagglutination inhibition antibody for Nescastle disease virus （ ND-HI ） in 7 - 14 day- old chickens and 60 -90 day-old young chickens were 2 -3 log2 higher than the control group after being vaccinated for 3 weeks, which could last for more than 4 months. The protection rate against avian pasteurella multocida was over 75.0% and its immune effect could last for 6 months. The field test of immune effect for duck and goose indicated that the titers of ND-HI antibody were all higher than 4.2 log2 in vaccinated ducks and geese while lower than 2 log2 in the control group after being vaccinated for 3 weeks. The protection rate against avian pasteurella multocida in vaccinated ducks and geese was higher than 75.0% and 62.5% respectively. [ Conclusion] The binary vaccine is safe for poultry and has good immune effects.

A Preliminary Study on Biological Activity of Wikstroemia chamaedaphne Meissn

[ Objective ] The insecticidal and antibacterial bioactivity of Wikstroemia chamaedaphne Meissn were screened and bioactive substances in it were separated and purified. [ Method] The Wikstroemia chamaedaphne Meissn was conducted ultrasonic extraction in petroleum ether, ethyl acetate and methanol. The insecticidal activity of Wikstroemia chamaedaphne Meissn to Mythimna separata walker and aphid were determined. The antibacterial activity of Wikstroemia chamaedaphne Meissn to Fusarium graminearu, Glomerella cingulata, F. oxysporium f. sp niveum, Alternaria solani and Fusarium oxysporium were also determined. The bioactivity-guided methods such as opencolumn chromatography and Pre-HPLC method were used to separate active components in petroleum ether extract from Wikstroemia chamaedaphne Meissn. [ Result] When the concentration was 500 mg/L, 3 kinds of extracts from Wikstroemia chamaedaphne Meissn didn＇ t show obvious antibacterial bioactivity to 5 kinds of test samples. When the concentration was 5%, petroleum ether extract show certain topical toxicity to aphids. The ethyl acetate extract showed certain antifeedant activity to 3^rd instar Larvae of Mythimna separata Walker. The fraction F4 of petroleum ether extract possessed highest topical toxicity to aphids and the lethality was 60.00%. [ Conclusion] Wikstroemia chamaedaphne Meissn contained many insecticidal constituents whose active parts and mechanism were needed further researches.

Expression of Recombinant Gene P1-2A3C of FMDV Asia I in Different Bombyx mori Varieties

[Objective] The aim was to investigate the expression level of recombinant gene P1-2A3C of FMDV Asia I in different Bombyx mori varieties,so as to screen out the B.mori varieties suitable for the foreign gene expression.[Method] The recombinant B.mori baculovirus rBmNPV（P1-2A3C） are injected into pupae of original species and hybrids of B.mori respectively.Then,the expression of antigen is detected by ELISA method and compared.[Result] The expression level of P1-2A3C significantly differed in different B.mori varieties,in which,the hybrids of Qiufeng×TQ78 and Qiufeng×Sijiaojian could be considered as the effective bioreactors for high-level expression of foreign genes.[Conclusion] This study provided the basis for breeding special B.mori varieties which can highly express target protein of Asia I FMDV.

Construction of Risk Assessment Application Model of Epidemic Disease in Large-scale Pig Farms

The application model of epidemic disease assessment technology for Web-based large-scale pig farm was expounded from the identification of epidemic disease risk factors, construction of risk assessment model and development of risk assessment system. The assessed pig farm uploaded the epidemic disease risk data information through on-line answering evaluating questionnaire to get the immediate evaluation report. The model could enhance the risk communication between pig farm veterinarian, manager and veterinary experts to help farm system understand and find disease risk factors, assess and report the potential high risk items of the pig farm in the three systems of engineering epidemic disease prevention technology, biological safety and immune monitoring, and promote the improvement and perfection of epidemic disease prevention and control measures.

Phylogenetic Analysis of the Sequences of rDNA Internal Transcribed Spacer (ITS) of Phytophthora sojae

The internal transcribed spacer （ITS） region （ITS1, ITS2 and 5.8S rDNA） of the nuclear ribosomal DNA （nrDNA） was amplified via the PCR method in seventeen different isolates of Phytophthora sojae using the common primers of the ITS of fungi. Around 800 bp- 1,000 bp fragments were obtained based on the DL2000 marker and the sequences of the PCR products were tested. Taking isolate USA as outgroup, the phylogenetic tree was constructed by means of maximum parsimony analysis, and the genetic evolution among isolates was analyzed. The results showed that there is a great difference between the base constitution of ITS 1 and ITS2 among various isolates. The seventeen isolates are classified into three groups, and the isolates from the same region belong to the same group, which shows the variation in geography.

Cloning and Sequence Analysis of HN and F Protein Genes from a Strain of Goose Paramyxovirus

[ Objective] The study was to clone HN and F genes from GX1 strain of goose paramyxovirus and analyze their sequences. [ Method] According to the full nucleotide sequence of GPV-SF02 strain of goose paramyxovirus, two pairs of pdmers were designed to amplify the HN and F genes from GX1 strain of goose paramyxovirus isolated from diseased goose in Guangxi Zhuang Autonomous Region; the amplified products were ligated into pMD18-T vector and sequenced. [ Result ] HN and F genes of this strain tested were 1 716 and 1 662 bp in full nucleotide length, respectively; both showed the homologues of about 97.3% with GPV- SF02 strain, of 80.3% -97.5% with strains LaSota, F48E9 and JS, of just 84.8% with Miyadera strain. [ Conclusion] The results show that isolated strain BX1 matches to virulent APMV-1 strain, belonging to genotype Ⅶ of APMV-1 strain.

Advances in Genetic Engineering Vaccine against Chicken Coccidiosis

Coccidiosis is a protozoan disease that seriously threatens the poultry in-dustry. At present, the prevention and treatment of chicken coccidiosis mainly rely on chemical drugs and coccidia live vaccine. However, drug control has problems of drug resistance and drug residue, and the coccidia live vaccine has drawbacks of pathogenicity, high production costs and virulence enhancement. Instead, genetic en-gineering vaccine is easy to operate, stable, safe and efficient, so it has become a hot spot in the prevention of chicken coccidiosis. In this paper, the research status, immune mechanism, immune effect＇s influencing factors and cytokine adjuvant appli-cation of genetic engineering vaccine against chicken coccidiosis were reviewed to provide some reference for the research and application of chicken coccidiosis gene vaccine.

Location of Highly Pathogenic Porcine Reproductive and Respiratory Syndrome Virus in Tissues of Natural Cases

[ Objective] The aim of this study was to provide a theoretical basis for the prevention and treatment of highly pathogenic porcine reproductive and respiratory syndrome （HP-PRRS）. [Method] Antigen location and histopathological observation in natural cases infected by highly pathogenic porcine reproductive and respiratory syndrome virus （HP-PRRSV） were analyzed by immunohistochemistry and H. E. staining. [Result] The virus antigen mainly existed in epithelial calls, and also a few in mecrophages, lymphocytes and brain nerve cells. [ Conclusion] The cell and tissue tropism of HP-PRRSV strain in natural cases is different from that of previous strains.

Antifungal Activity of Botanical Extracts against Botrytis cinerea and Alternaria solani

[Objective] The antifungal activity of the extracts from,Atractylodes macracephal Koidz and Pulsatilla chinensis Bunge Regel,against Botrytis cinerea and Alternaria solani were studied under the condition of laboratory,in order to develop and utilize these two plants.[Method] The mycelium growth rate test was applied to measure the antifungal activities of extracts against fungi.[Result] the extracts of all the two plants showed strong antifungal activity against the target pathogenic fungi,especially the antifungal activity of the extract from Pulsatilla chinensis Bunge Regel was stronger and more stable.The inhibition rate to the mycelium growth of Botrytis cinerea was 80.25%.At the same concentration,the extract from Atractylodes macracephal Koidz showed little inhibition to Botrytis cinerea and Alternaria solani.The petrolelum ether extract of Atractylodes macracephal Koidz showed stronger antifungal activities and the EC50 was 5.31 mg/ml,and the n-butanol extract of Pulsatilla chinensis Bunge Regel showed stronger antifungal activities and the EC50 was 2.93 mg/ml.[Conclusion] The extracts from Pulsatilla chinensis Bunge Regel showed the stronger antifungal activity against Botrytis cinerea and Alternaria solani.

Inhibitory Effects of Some Plant Extracts on 5 Bacterial Strains

[Objective] This study was conducted to develop potential natural plant products for controlling walnut blight pathogen and other bacteria. [Method] Inhibitory effects of extracts obtained from 15 plants with 3 solvents on bacteria were investi- gated by disk diffusion method. [Results] Except the extracts from Magnolia grandi- flora and Typha orientalis, extracts of 13 plant leaves presented inhibitory effects on 5 bacteria strains to certain degrees. Among them, the effect of water extract of Aesculu schinensis on Bacillus sp. XHE8 was the strongest, with inhibition zone di- ameter reaching （31.3＋3.9） mm and the ratio to control above 5.0. Four of the 5 tested strains were sensitive to the extracts of Sambucus chinensis, and 3 of them were inhibited by Ophiopegon japonicas extracts and Reineckia camea extracts, with ratios of treatment to control large than 1.5 in all. Leaf extract of A. chinensis had significant anti-bacteria ability, and could be used as a potential plant source for bactericide. [Conclusion] The results laid a foundation for exploring active com- pounds and elucidating the mechanism in it.