Association of circulating levels of soluble PD-1, PD-1 gene polymorphisms with HBV infection and HBV infection-associated hepatocellular carcinoma / 中华预防医学杂志

Objective: To investigate the association of circulating sPD-1 level and PD-1 gene polymorphisms with HBV infection and HBV infection-associated hepatocellular carcinoma. Methods: A case-control study was conducted. A total of 237 chronic HBV infection cases and 138 HBV infection-associated hepatocellular carcinoma in the Department of Infectious Diseases of the First Hospital of Shanxi Medical University from 2018 to 2021 were selected as the case group. About 250 individuals who visited a hospital physical examination center for routine physical examination during the same period were selected as the control group. Plasma sPD-1 levels were measured by using an ELISA kit and genotyping was performed by using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique. The association of sPD-1 levels and PD-1 polymorphisms with HBV infection as well as HBV infection-associated hepatocellular carcinoma was analyzed by using logistic regression models after adjusting for age, sex, alcohol consumption, smoking, ALT and AST levels. The sPD-1 level and PD-1 polymorphisms were independent variables, and HBV infection was the dependent variable. Results: The age of 237 chronic HBV infections, 138 HBV infection-related liver cancer case subjects and 250 control subjects in the study was (49.1±10.8), (51.9±12.7) and (50.7±11.9) years, respectively. Multivariate logistic regression model analysis showed that with a 1 pg/ml increase in sPD-1 level, the OR (95%CI) values for the risk of incident HBV infection cases and HBV hepatocellular carcinoma cases were 1.92 (1.68-2.19) and 2.02 (1.69-2.40). For rs2227981, compared with the CC genotype, the TT genotype had a lower risk of HBV infection and liver cancer associated with HBV infection, with OR (95%CI) values of 0.45 (0.22-0.91) and 0.35 (0.14-0.91). For rs2227982, compared with the CC genotype, the CT and TT genotypes also had a lower risk of HBV infection [OR (95%CI) values of 0.72 (0.53-0.97) and 0.57 (0.35-0.93)] and HBV infection-related liver cancer [OR (95%CI) values of 0.64 (0.45-0.92) and 0.52 (0.29-0.93)]. Conclusions: Plasma sPD-1 levels and PD-1 gene polymorphisms are associated with HBV infection and HBV infection-associated hepatocellular carcinoma.

Correlation between intuitive eating and self rated health among college students / 中国学校卫生

Objective@#To investigate average score and correlation of IES-2 and Self rated Health Measurement Scales V 1.0 (SRHMS V 1.0 ) among Chinese college students, so as to provide a scientific reference for health promotion of college students.@*Methods@#A random cluster sampling method was applied to conduct an online questionnaire survey from July to December 2019 among 542 college students from 8 universities. Multiple linear regression was used to analyze the association of intuitive diet level on self reported health status after adjusting for confounding factors.@*Results@#The average scores of SRHMS V1.0 was (69.84± 10.28 ), and the mean scores of physical self rated health (PSH), mental selfrated health (MSH) and social self rated health (SSH) sub scale were(78.50±10.39)(61.86±14.53)(67.54±14.71), respectively. After adjusting for confounders, the β coefficient of IES-2 and SRHMS V1.0, PSH, MSH and SSH were 6.46, 5.00,10.15 and 3.90 ( P <0.05). Higher level of EPR had positive effects on SRHMS V1.0, PSH and MSH ( β =2.47,2.30,4.71, P <0.05). Higher level of RHSC had positive effects on SRHMS V1.0 , PSH, MSH and SSH( β =2.44, 1.69, 2.71，3.16, P <0.05).Higher level of B-FCC had positive effects on SRHMS V1.0, PSH, MSH and SSH( β =3.71,2.53,4.68，4.17, P <0.05).@*Conclusion@#The intuitive eating has a positive effect on overall health, especially on mental health. College students should avoid emotional eating, and eat in response to hunger and satiety signals as well as eat healthy food to meet physical needs when facing physical, mental and social problems.

Polymorphism of P66 in / 生物医学与环境科学（英文）

Objective@#To study the polymorphism in P66 and its human B-cell epitopes of @*Methods@#Polymerase chain reaction (PCR) and sequencing were used to obtain the P66 sequences of 59 Chinese @*Results@#Results showed that genetic and amino acid diversity presented in the 66 kD protein of all 59 Chinese strains, especially in @*Conclusion@#In P66 of 59 Chinese strains, polymorphisms were widely distributed. More importantly, the P66 amino acid sequences of

MicroRNA-34c promotes neuronal recovery in rats with spinal cord injury through the C-X-C motif ligand 14/Janus kinase 2/signal transducer and activator of transcription-3 axis / 中华医学杂志(英文版)

BACKGROUND@#Developing effective spinal cord repair strategies for spinal cord injury (SCI) is of great importance. Emerging evidence suggests that microRNAs (miRNAs) are closely linked to SCI recovery. This study aimed to investigate the function of miR-34c in the neuronal recovery in rats with SCI.@*METHODS@#A rat model with SCI was established. Differentially expressed miRNAs were identified by a microarray analysis. MiR-34c expression in rats was measured by reverse transcription quantitative polymerase chain reaction. Altered expression of miR-34c or C-X-C motif ligand 14 (CXCL14) was introduced in SCI rats to measure their roles in neuronal recovery. Western blot analysis was performed to determine the phosphorylation of Janus kinase 2 (JAK2) and signal transducer and activator of transcription-3 (STAT3). Neuronal apoptosis in rat spinal cord tissues was detected. The concentrations of SCI recovery-related proteins thyrotropin releasing hormone (TRH), prostacyclin (PGI2), and ganglioside (GM) were evaluated by enzyme-linked immunosorbent assay. Data were analyzed using a t-test with a one-way or two-way analysis of variance.@*RESULTS@#Rats with SCI presented decreased grip strength (112.03 ± 10.64 vs. 17.32 ± 1.49 g, P < 0.01), decreased miR-34c expression (7 days: 3.78 ± 0.44 vs. 0.95 ± 0.10, P < 0.05), and increased CXCL14 expression (7 days: 0.61 ± 0.06 vs. 2.91 ± 0.27, P < 0.01). MiR-34c was found to directly bind to CXCL14. Overexpression of miR-34c increased grip strength (11.23 ± 1.08 vs. 31.26 ± 2.99 g, P < 0.01) and reduced neuronal apoptosis in spinal cord tissues (53.61% ± 6.07% vs. 24.59% ± 3.32%, P < 0.01), and silencing of CXCL14 also increased the grip strength (12.76 ± 1.13 vs. 29.77 ± 2.75 g, P < 0.01) and reduced apoptosis in spinal cord tissues (55.74% ± 6.24% vs. 26.75% ± 2.84%, P < 0.01). In addition, miR-34c upregulation or CXCL14 downregulation increased the concentrations of TRH, PGI2, and GM, and reduced phosphorylation of JAK2 and STAT3 in rats with SCI (all P < 0.01).@*CONCLUSION@#The study provided evidence that miR-34c could promote neuronal recovery in rats with SCI through inhibiting CXCL14 expression and inactivating the JAK2/STAT3 pathway. This study may offer new insights into SCI treatment.

Molecular cloning and expression of OspA peptide from a Chinese Borrelia garinii strain PD91 and preliminary study on its immunoprotectivity / 中华微生物学和免疫学杂志

Objective:To clone and express the 126-274 aa OspA peptide (OspA-pep) of Chinese Borrelia garinii ( B. garinii) strain PD91 and to preliminarily study its immune protectivity. Methods:The gene encoding the 126-274 aa OspA-pep of B. garinii PD91 was amplified by polymerase chain reaction (PCR) and then cloned into the prokaryotic expression vector pET-30a to construct the recombinant plasmid pET-30a-OspA-pep. Escherichia coli BL21 (DE3) competent cells transfected with the recombinant plasmid were induced by IPTG to express the target protein. The recombinant OspA-pep (rOspA-pep) was purified with Ni-IDA resin chromatography and its immunogenicity was analyzed by Western blot. New Zealand white rabbits were immunized with different doses of rOspA-pep (20, 30, 40, 50, 60, 80 and 100 μg). The titers of specific IgG antibodies in rabbit serum samples before and after immunization were detected by indirect immunofluorescence assay (IFA). The optimal immune dose was determined according to the antibody titer after immunization. In vitro neutralization test was performed to detect the immune protection of rOspA-pep using serum samples of the optimal immunization group. The optimal dose of rOspA-pep was used to immunize New Zealand white rabbits to observe the changes in antibody titer. Results:The recombinant plasmid pET-30a-OspA-pep was successfully constructed and highly expressed in host bacteria. Western blot showed that rOspA-pep had obvious antigen-antibody reaction with polyclonal antibody against B. garinii PD91 strain. IFA results showed the titers of IgG antibody in serum samples of rabbits immunized with rOspA-pep increased significantly (up to 1∶2 480) and 40 μg was the optimal dose. The neutralization rates of antibodies induced by 40 μg of rOspA-pep were 100% against 10 6 strain/ml of representative B. garinii PD91 and Borrelia afzelii ( B. afzelii) FP1 strains, 100% against 10 7 strain/ml of FP1 strain, and 60% against 10 7 strain/ml of PD91 strain. After immunization with 40 μg rOspA-pep on 1 d and 30 d, the titers of specific IgG antibody in rabbit serum samples reached the peak within two months, and maintained at that level for about 3-4 months before a gradual decline. Conclusions:The 126-274 aa OspA peptide fragment of Chinese B. garinii PD91 strain possessed good immunogenicity and induced antibodies with better in vitro neutralizing activity, which suggested that it could be used as a candidate component of the second generation subunit vaccine in China.

Chinese Herbal Formula Ermiao Powder () Regulates Cholinergic Anti-inflammatory Pathway in Rats with Rheumatoid Arthritis / 中国结合医学杂志

OBJECTIVE@#To investigate the effect of Chinese herbal formula Ermiao Powder (, EMP) on the expression of cholinergic anti-inflammatory pathway in rats with rheumatoid arthritis (RA).@*METHODS@#Seventy-two rats were randomly divided into 6 groups according to body weight, including normal control group, collageninduced arthritis (CIA) group, three doses EMP groups, and methotrexate (MTX) group (n=12 per group). All of the rats except for those in the normal control group were given multipoint subcutaneous injection of bovine type II collagen to establish a CIA model. Three EMP groups received a high- (4.5 g/kg), medium- (3.0 g/kg), and low- (1.5 g/kg) doses of EMP by intragavage, respectively. MTX group was injected intraperitoneally MTX at 0.9 mg/kg once a week as the positive control. The administration was 3 consecutive weeks. Joint swelling, arthritis index, and body weight changes in different experimental groups of rats were tested. The joint damage was evaluated by masson staining. Quantitative real-time polymerase chain reaction, Western blot, and immunohistochemistry (IHC) were performed to evaluate the expression of CHRNA7, encoding α7 nicotinic acetylcholine receptor in the cholinergic anti-inflammatory pathway, in different tissues and their localization in the spleen and joints.@*RESULTS@#CHRNA7 expression levels were significantly higher in the joints and spleens of CIA group than those in normal control group (both P<0.05). Moreover, the CHRNA7 mRNA and protein levels in the spleen and joints of MTX and three doses of EMP groups were significantly lower than CIA group (all P<0.05). Compared with the MTX group, treatment with low-dose EMP resulted in significant reduction of CHRNA7 mRNA and protein expression levels (P<0.05 or P<0.01). IHC showed positive signals of CHRNA7 in the white pulp and red pulp of the spleens of rats; CHRNA7 was expressed on fibroblast-like synoviocytes, macrophages, and endothelial cells in the joints of rats, and the expression in the joints of low-dose EMP group was significantly lower than that in the CIA group (P<0.01).@*CONCLUSIONS@#Cholinergic anti-inflammatory pathway was involved in the generation of the inflammatory reaction in CIA rats, and EMP exerted therapeutic effect on RA through cholinergic anti-inflammatory pathway.

Quality Relevance of Standard Decoction and Different Drying Methods Products for Gentianae Macrophyllae Radix Granules Based on HPLC Fingerprint / 中国实验方剂学杂志

Objective:To establish the fingerprints of standard decoction of Gentianae Macrophyllae Radix-dried products by different methods,and to evaluate the quality correlation. Method:HPLC,InertSustain C18 chromatographic column(4.6 mm×250 mm,5 μm),Gradient elution was performed for the mobile phase of acetonitrile-phospho,detection wavelength at 240 nm and flow rate of 0.8 mL·min-1,and column temperature was 40℃. The quality correlation analysis of different methods for different kinds of Gentianae Macrophyllae Radix standard decoction was carried out from the aspects of chemical composition consistency,common chemical composition consistency,main chemical composition content and transfer rate. Result:The control fingerprint of Gentianae Macrophyllae Radix standard decoction was established. According to the peak matching data,there were 10 common peaks in the fingerprint of 15 batches of Gentianae Macrophyllae Radix standard decoction.Among the 10 common peaks,5 chemical constituents of loganic acid,6'-O-β-D-glucosyl gentiopicroside,swertiamarin,gentiopicroside and swertia glycosides were identified. The results of quality correlation analysis showed that the three different drying methods were consistent with the chemical composition and quantity of Gentianae Macrophyllae Radix standard decoction. But in terms of the content consistency of common chemical components and the transfer rate of main chemical components,the quality correlation between the products obtained from vacuum drying and the standard decoction was lower than that obtained from spray drying and freeze-drying. Conclusion:The fingerprint of different method of Gentianae Macrophyllae Radix standard decoction was established. Through the analysis of the mass correlation of chemical composition consistency,common chemical composition content consistency,main chemical composition content and transfer rate,the mass correlation between them was comprehensively reflected. It is suggested that spray drying or freeze drying should be used for the key drying process of Gentianae Macrophyllae Radix granules. This study provides a reference for the preparation process and quality control of Gentianae Macrophyllae Radix granules.

Clinical observation of autologous penetrating keratoplasty in the treatment of corneal blindness / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To investigate the clinical effect of autologous penetrating keratoplasty in the treatment of corneal blindness. <p>METHODS: Totally 8 patients with corneal blindness were treated by autologous penetrating keratoplasty in our hospital from 2014-01 to 2018-03. Using retrospective analysis, the patients were followed up for one year. To observe the intraoperative complications and postoperative conditions such as visual acuity, corneal transparency, and other were observed.<p>RESULTS: The uncorrected visual acuity of all the 8 patients was greater than 0.02 1wk after operation, and the rate of restoration of visual acuity was 100%(8/8). The corrected visual acuity of 5 patients(5-8)was more than 0.3 1mo after operation. The corrected visual acuity of 3 patients(3-8)with severe cataract before operation was improved to 0.08-0.2. One year later, all the corneal grafts in the recipient eyes were transparency and no recurrence of infection or secondary infection occurred in all 8 patients. <p>CONCLUSION: Corneal graft is easy to grow and there are no exclusion reactions, fewer postoperative complications after autologous corneal transplantation. So, corneal implants can remain transparent for a long time, and the surgery cost is lower. Autologous corneal transplantation can not only provide long-term useful vision for patients with monocular blindness combined with corneal blindness, but also reduce their financial burden and bring great benefits to patients.

Preparation and Evaluation of the Serum Quality Control Material of β2-Microglobulin / 现代检验医学杂志

Objective To prepare quality control material of β2-microglobulin (β2 MG),then evaluate the quality of it.Methods The quality control material of β2-MG was prepared with pooled serum from patients.Then according to Guidance on Evaluating the Homogeneity and Stability of Samples Used for Proficiency Testing and ISO Guide 35,the homogeneity and stability of it was evaluated.Results According to the result of homogeneity test,there was no statistically significant difference between within groups and between-groups (F=0.699,2.091,all P＞0.05) for 2 levels of quality control materi al.What was more,the stability test showed that it could stabilize for at least 5 days at 25 ℃ (t=-1.76,-2.64,all P＞ 0.05),28 days at 4 ℃ (t=-1.86,-2.44,all P＞0.05),4 months at-20 ℃ (t=0.26,-2.68,allP＞0.05),and 1 yearat -80℃ (t=-0.96,0.01,all P＞0.05).Conclusion Quality control material of β2-MG has simple preparation.The homogeneity and stability of it meets the standard requirements.

Ocular surface evaluation among exotropia patients in different age groups / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To investigate the ocular surface condition among exotropia patients of different age groups by Oculus anterior segment analyzer. METHODS: The ocular surface condition of 66 patients with extropia were examined by Oculus anterior segment analyzer, including tear meniscus height, tear meniscus height after irritation,redness index,non-invasive break-up time(BUT),average BUT. Three groups were divided according to age:2-7 years,8-18 years and 19-46 years. RESULTS: BUT, temporal conjunctival redness index, temporal ciliary redness index,nasal ciliary redness index of 2-7 years group were statistically different to those of 8-18 years group and 19-46 years group. There were no statistically differentiations between redness index at different areas in each group and no statistically differentiations in redness index between dominant eye and nondominant eye(P>0.05). Redness index showed a positive lineal correlation with age. CONCLUSION:Patients are combined with dry eye and unstable tear firm beside strabismus before surgery. Ocular surface assessment should be paid attention to in preschool children in order to prevent complications around operation. Ocular surface data of children with different ages needs to be screened and collected in China.

Palliative surgery for refractory purulent corneal ulcer / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM:To investigate the clinical effect of scleral allograft combined with conjunctival flap autograft for refractory purulent corneal ulcer. <p>METHODS: Twenty patients(20 eyes)with purulent corneal ulcer admitted to our hospital during June 2015 to June 2017 were selected. The results showed that the drug treatment was not effective and could not be performed by penetrating keratoplasty, and then allogeneic scleral transplantation combined with conjunctival flap autograft covering was performed. A retrospective analysis was carried out. The intraoperative complications, postoperative scleral and conjunctival flap growth and postoperative complications of this palliative operation were observed. <p>RESULTS: Of the 20 patients, 18 had good growth of sclera and conjunctival flap, the cure rate was 90%. One case had bad healing of conjunctival flap and was cured by amniotic membrane transplantation; 1 case had enucleation because of severe vitreous cavity infection; 2 cases had lens prolapse, the incidence rate was 10%. Secondary glaucoma occurred in 3 cases with an incidence of 15%, which was cured by cryociliary surgery.<p>CONCLUSION:Scleral allograft combined with conjunctival flap autograft is a palliative operation. It is an effective way to control the corneal infection and maintain the appearance of the eyeball in the patients with severe purulent corneal ulcer, which is ineffective in drug treatment and unable to perform penetrating keratoplasty. It can replace the previous enucleation of the eye contents to control the infection. It can not only avoid the eye pain and psychological trauma caused by the loss of eyeball, but also save money for improving the appearance in the later stage.

Evaluation of Six Recombinant Proteins for Serological Diagnosis of Lyme Borreliosis in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>In this study, we evaluated the diagnostic efficiency of six recombinant proteins for the serodiagnosis of Lyme borreliosis (LB) and screened out the appropriate antigens to support the production of a Chinese clinical ELISA (enzyme-linked immunosorbent assay) kit for LB.</p><p><b>METHODS</b>Six recombinant antigens, Fla B.g, OspC B.a, OspC B.g, P39 B.g, P83 B.g, and VlsE B.a, were used for ELISA to detect serum antibodies in LB, syphilis, and healthy controls. The ELISA results were used to generate receiver operating characteristic (ROC) curves, and the sensitivity and specificity of each protein was evaluated. All recombinant proteins were evaluated and screened by using logistic regression models.</p><p><b>RESULTS</b>Two IgG (VlsE and OspC B.g) and two IgM (OspC B.g and OspC B.a) antigens were left by the logistic regression model screened. VlsE had the highest specificity for syphilis samples in the IgG test (87.7%, P<0.05). OspC B.g had the highest diagnostic value in the IgM test (AUC=0.871). Interactive effects between OspC B.a and Fla B.g could reduce the specificity of the ELISA.</p><p><b>CONCLUSION</b>Three recombinant antigens, OspC B.g, OspC B.a, and VlsE B.a, were useful for ELISAs of LB. Additionally, the interaction between OspC B.a and Fla B.g should be examined in future research.</p>

Prediction of potential geographic distribution of Lyme disease in Qinghai province with Maximum Entropy model / 中华流行病学杂志

Objective To predict the potential geographic distribution of Lyme disease in Qinghai by using Maximum Entropy model (MaxEnt).Methods The sero-diagnosis data of Lyme disease in 6 counties (Huzhu,Zeku,Tongde,Datong,Qilian and Xunhua) and the environmental and anthropogenic data including altitude,human footprint,normalized difference vegetation index (NDVI) and temperature in Qinghai province since 1990 were collected.By using the data of Huzhu Zeku and Tongde,the prediction of potential distribution of Lyme disease in Qinghai was conducted with MaxEnt.The prediction results were compared with the human sero-prevalence of Lyme disease in Datong,Qilian and Xunhua counties in Qinghai.Results Three hot spots of Lyme disease were predicted in Qinghai,which were all in the east forest areas.Furthermore,the NDVI showed the most important role in the model prediction,followed by human footprint.Datong,Qilian and Xunhua counties were all in eastern Qinghai.Xunhua was in hot spot area Ⅱ,Datong was close to the north of hot spot area Ⅲ,while Qilian with lowest sero-prevalence of Lyme disease was not in the hot spot areas.The data were well modeled in MaxEnt (Area Under Curve=0.980).Conclusions The actual distribution of Lyme disease in Qinghai was in consistent with the results of the model prediction.MaxEnt could be used in predicting the potential distribution patterns of Lyme disease.The distribution of vegetation and the range and intensity of human activity might be related with Lyme disease distribution.

Combination of Loop-Mediated Isothermal Amplification Assay and Nested PCR for Detection of Borrelia burgdorferi sensu lato in Human Serum Samples / 生物医学与环境科学（英文）

A set of universal loop-mediated isothermal amplification (LAMP) primers targeting the fla gene was designed to detect Borrelia burgdorferi sensu lato (B. burgdorferi s.l.) in human samples. The sensitivity of LAMP was 20 copies/reaction, and the assay did not detect false positives among 11 other related bacteria. A positive LAMP result was obtained for 9 of the 24 confirmed cases and for 12 of 94 suspected cases. The positive rate of LAMP was the same as that of nested PCR. The LAMP is a useful diagnostic method that can be developed for rapid detection of B. burgdorferi s.l. in human sera. Combination of the LAMP and nested PCR was more sensitive for detecting B. burgdorferi s.l. in human serum samples.

Molecular cloning and expression of OspC protein of a Chinese Borrelia afzelli FP1 strain and pre-liminary study on the immune protectivity of the rOspC protein / 中华微生物学和免疫学杂志

Objective To clone and express the outer surface protein C ( OspC) from a Chinese Borrelia afzelli FP1 strain and to evaluate the immune protectivity of the recombinant OspC protein ( rOspC) . Methods The gene encoding OspC protein of Borrelia afzelli FP1 strain was amplified by polymerase chain reaction (PCR) and then inserted into pET-30a plasmid to construct the recombinant expression plasmid pET-30a-OspC. The transformed E. coli BL21 strains carrying pET-30a-OspC plasmid were induced by IPTG to express OspC protein. The expressed proteins were purified by Ni-IDA resin chromatography and analyzed by SDS-PAGE and Western blot assay. Indirect immunofluorescence assay ( IFA) was performed to detect anti-rOspC protein antibodies in serum samples from rabbits immunized with rOspC protein. In vitro neutral-ization test was performed for evaluation the immune protectivity of rOspC protein. Results The recombi-nant expression plasmid pET-30a-OspC was successfully constructed and highly expressed in E. coli BL21. A strong antigen-antibody reaction between the rOspC protein and polyclonal antibody against Borrelia afzelli FP1 strain was detected by Western blot assay. The titers of IgG in serum samples from rabbits immunized with rOspC protein were significantly elevated. The in vitro neutralization test indicated that 106/ml of Borre-lia afzelli FP1 strains were neutralized by every anti-OspC protein serum sample from the experiment group. Conclusion The rOspC protein showed a strong immune protectivity against Borrelia afzelli, which could be used in the development of polyvalent subunit vaccine against lyme disease.

Nested-PCR and LAMP in detection of B .burgdor f eri in ticks / 中国人兽共患病学报

ABSTRACT:Nested‐PCR and loop‐mediated isothermal amplification (LAMP) were applied to identify the Borrelia burg‐dor f eri (B .burgdor f eri) in ticks in this study .A total of 112 adult ixodes were collected from vegetation in a forest area and farm animals in Xunhua County ,Qinghai Province and Xinbin County ,Liaoning Province .The ticks were examined for the presence of B .burgdorferi by nested‐PCR and LAMP .Results showed that 12 in 51 samples were found positive in Xunhua County (23 .53% ) .While positive rate in Xinbin County was 29 .51% with 18 samples positive in 61 samples .In total of 112 tick samples ,the PCR‐positive rate was 17 .86% with 20 positive samples identified ,whereas 15 positive samples were con‐firmed with positive rate of 13 .39 % by LAMP assay .There was no significant difference between the two assays (Х2 =0 .85 , P>0 .05) .Results suggest that both nested‐PCR and LAMP could be used in identifying B .burgdorferi in ticks .Combina‐tion of these two assays could improve the testing results .This is the first report of B .burgdorferi in ticks in Xunhua and Xinbin counties ,and helps to complete the database of the infection rate of B .burgdor f eri in ticks in the widely‐forested area of China .

Establishment of multiple locus variable-number tandem repeat analysis assay for genotyping of Borrelia burgdorferi sensu lato detected in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Human Lyme Borreliosis (LB), which is caused by Borrelia burgdorferi sensu lato (B. burgdorferi), has been identified as a major arthropod-borne infectious disease in China. We aimed to develop a multiple locus variable-number tandem repeat (VNTR) analysis (MLVA) assay for the genotyping of Borrelia burgdorferi strains detected in China.</p><p><b>METHODS</b>B. garinii PBi complete 904.246 kb chromosome and two plasmids (cp26 and lp54) were screened by using Tandem Repeats Finder program for getting potential VNTR loci, the potential VNTR loci were analyzed and identified with PCR and the VNTR loci data were analyzed and MLVA clustering tree were constrcted by using the categorical coefficient and the unweighted pair-group method with arithmetic means (UPGMA).</p><p><b>RESULTS</b>We identified 5 new VNTR loci through analyzing 47 potential VNTR loci. We used the MLVA protocol to analyse 101 B. burgdorferi strains detected in China and finally identified 51 unique genotypes in 4 major clusters including B. burgdorferi sensu stricto (B.b.s.s), B. garinii, B. afzelii, and B. valaisiana, consistent with the current MLSA phylogeny studies. The allele numbers of VNTR-1, VNTR-2, VNTR-3, VNTR-4, and VNTR-5 were 7, 3, 9, 7, and 6. The Hunter-Gaston index (HGI) of five VNTR loci were 0.79, 0.22, 0.77, 0.71, and 0.67, respectively. The combined HGI of five VNTR loci was 0.96. Clustering of the strains of Xinjiang, Inner Mongolia and Heilongjiang was confirmed, and this situation was consistent with the close geographical distribution of those provinces.</p><p><b>CONCLUSION</b>The MLVA protocol esytablished in this study is easy and can show strains' phylogenetic relationships to distinguish the strains of Borrelia species. It is useful for further phylogenetic and epidemiological analyses of Borrelia strains.</p>

Seroepidemiological investigation of lyme disease and human granulocytic anaplasmosis among people living in forest areas of eight provinces in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Lyme disease and Human granulocytic anaplasmosis are tick-borne diseases caused by Borrelia burgdorferi and Anaplasma phagocytophilum respectively. We have investigated infection and co-infection of the two diseases in the population of forest areas of eight provinces in China by measuring seroprevalence of antibodies against B. burgdorferi and A. phagocytophilum.</p><p><b>METHODS</b>Forest areas in 8 provinces were chosen for investigation using whole sampling and questionnaire survey methods. 3 669 serum samples from people in the forest areas were tested for the presence of antibodies by indirect immunofluorescent assay (IFA).</p><p><b>RESULTS</b>Seroprevalence against B. burgdorferi was 3% to 15% and against A. phagocytophilum was 2% to 18% in the study sites in the 8 provinces in China. We also found co-infection of B. burgdorferi and A. phagocytophilum in 7 of the 8 provinces (the exception being the Miyun area in Beijing). The seroprevalence for both B. burgdorferi and A. phagocytophilum was significantly higher among people exposed to ticks than among people who were not exposed to ticks.</p><p><b>CONCLUSION</b>We conclude that both pathogens are endemic in the forest areas in the eight provinces, but the prevalence of B. burgdorferi and A. phagocytophilum differs between the provinces.</p>

A study of the technique of western blot for diagnosis of lyme disease caused by Borrelia afzelii in China / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To study the technique of Western blot for the diagnosis of Lyme disease caused by Borrelia afzelii in China and to establish the standard criteria by operational procedure.</p><p><b>METHODS</b>FP1, which is the representative strain of B. afzelii in China, was analyzed by SDS-PAGE, electro transfer and immunoblotting assays. The molecular weights of the protein bands of FP1 were analyzed by Gel-Pro analysis software. In a study using 451 serum samples (159 patients with Lyme disease and 292 controls), all observed bands were recorded. The accuracy of the WB as a diagnostic test was established by using the ROC curve and Youden index.</p><p><b>RESULTS</b>Criteria for a positive diagnosis of Lyme disease were established as at least one band of P83/100, P58, P39, OspB, OspA, P30, P28, OspC, P17, and P14 in the IgG test and at least one band of P83/100, P58, P39, OspA, P30, P28, OspC, P17, and P41 in the IgM test. For IgG criteria, the sensitivity, specificity and Youden index were 69.8%, 98.3%, and 0.681, respectively; for IgM criteria, the sensitivity, specificity and Youden index were 47%, 94.2%, and 0.412, respectively.</p><p><b>CONCLUSION</b>Establishment of WB criteria for B. afzelii is important in validating the diagnostic assays for Lyme disease in China.</p>

Optimization of pulse-field gel electrophoresis for Borrelia burgdorferi subtyping / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>To optimize the performance of Pulsed-Field Gel Electrophoresis (PFGE) for the comparison of inter-laboratory results and information exchange of Borrelia burgdorferi subtyping.</p><p><b>METHODS</b>A panel of 34 strains of B. burgdorferi were used to optimize PFGE for subtyping. In order to optimize the electrophoretic parameters (EPs), all 34 strains of B. burgdorferi were analyzed using four EPs, yielding different Simpson diversity index (D) values and the epidemiological concordance was also evaluated.</p><p><b>RESULTS</b>The EP of a switch time of 1 s to 25 s for 13 h and 1 s to 10 s for 6 h produced the highest D value and was declared to be optimal for MluI and SmaI PFGE of B. burgdorferi. MluI and SmaI were selected as the first and second restriction enzymes for PFGE subtyping of B. burgdorferi according to discrimination and consistency with epidemiological data.</p><p><b>CONCLUSION</b>PFGE can be used as a valuable test for routine genospecies identification of B. burgdorferi.</p>