Cultivation of murine thymic epithelium has been established.Murine thymic epi-thelial cultural supernatant showed activity increasing both spontaneous ~3H-TdR inco-rporation and ConA-stimulated proliferation of thymocytes by the exponent of 1.5-2at dilution 1:8 to 1:32,the peak of the activity was present at 10-20 or 15 days ofcultivation.However,the thymic epithelial supernatant from cultures at 6-7 daysshowed activity suppressing both proliferations of thymocytes at dilution 1:8-1:16.Theresult showed thymic epithelial supernatant had no 1L-2 activity.

Objective To assess willingness to pay(wtp)for typhoid vi vaccine in typhoid epidemic area in Guangxi Zhuang Autonomous Region and to provide evidence for introduction of the vaccine.Methods Applying the method of wtp was investigated in typhoid epidemic area,the curve of "price-accept ratio model"was drawn up with Probit regression.Results The formula of "price-accept ratio model" was described as following:Probit(p)=0.88952-0.46296X.The WTP for typhoid vi vaccine was $10.41,with a 95% confidence interval of $6.67～16.24.Conclusions WTP for typhoid vi vaccine was around $10 in Guangxi typhoid epidemic area.The Contingent Valuation Method was applicable in the investigation of WTP.

Objective To study the morphological changes in peripheral blood cells and bone marrow cells of the patients suffering from mustard gas poisoning and their relationship with the degree of poisoning. Method The peripheral blood cells and marrow cells were examined morphologically after mustard gas poisoning. Results The total WBC count was reduced progressively in 44 patients, among them 10 patients showed granulocytopenia or agranulocytosis. Severe injury to eosinophils was seen also in the early period, and the percentage and absolute value of lymphocytes were significantly lowered too. However, platelets were not significantly influenced. The red cell count and hemoglobin level were elevated because of hemoconcentration. Blood marrow cells showed marked morphological changes. WBC became swollen with appearance of poisonous granules and vacuoles in their cytoplasm. Deformed lymphocytes (LY) could be found in the marrow cells of all the patients, mainly in the form of immature lymphocytes. It was shown that mustard gas produced serious damage to the hematopoitic cells of the bone marrow in the early period. Conclusions The examination of pheripheral blood and bone marrow cells is an important measure to determine the degree of mustard gas poisoning. The decrease in the absolute value of lymophocyte count and the time of its recovery, and degree of inhibition of hematopoiesis are closely related to the degree of mustard gas poisoning.

ObjectiveTo investigate the efficacy of mountain-burning fire acupunctureat point Yongquan(KI1)in treating spinal cord injury.MethodThirty patients with spinal cord injury were randomly allocated to observation and control groups, 15 cases each. The control group received modern rehabilitation therapy and the observationgroup, mountain-burning fire acupuncture at point Yongquan plus exercise rehabilitation therapy.ResultThe total efficacy rate was 93.3% in the observation group and 73.3% in the control group. The total efficacy rate increased significantly in the observation group and was significantly higher than in the control group(P<0.05).ConclusionMountain-burning fire acupuncture at point Yongquan has a marked therapeutic effect on spinal cord injury and effectively improves urinary and fecal voiding, sexual and limb motor functions in the patients.

Objective:To understand whether there are drug resistant and disinfectant resistant Yersinia pestis strains in China, and to provide accurate information for clinical treatment of plague. Methods:A total of 2 753 Yersinia pestis strains isolated from 10 natural plague foci in China from 1943 to 2016 were collected. According to National Center for Biotechnology Information (NCBI) released sequences of aminoglycoside streptomycin resistant genes strA, strB, β-lactam antibiotics resistant genes TEM, SHV and CTX-M, sulfamilamide resistant genes sul1, sul2 and sul3, and disinfectant resistant gene qacE△1-sul1, a pair of primers of each gene was designed for above-mentioned genes. Genomic DNA of 2 753 strains of Yersinia pestis was extracted, and the 9 target genes of all DNA samples were amplified by PCR. Results:Negative and positive controls of PCR detection were established. No corresponding target bands of aminoglycoside streptomycin resistant genes strA, strB, β-lactam antibiotics resistant genes TEM, SHV and CTX-M, sulfamilamide resistant genes sul1, sul2 and sul3, and disinfectant resistant gene qacE△1-sul1 were found in the DNA samples of 2 753 strains of Yersinia pestis.Conclusion:The above-mentioned genes of drug resistance and disinfectant resistance have not been detected in Yersinia pestis of China, but the monitoring of drug resistance of Yersinia pestis still needs to be carried out continuously.

Objective To establishment a method for detection of multiple drug resistance gene of Yersina pestis using polymerase chain reaction(PCR), to provide a guidance for treatment of plague. Methods According to National Center for Biotechnology Information (NCBI) released sequences of aminoglycoside resistant genes of streptomycin resistant,strB,strA,beta lactam antibiotics resistant genes tem,shv,and ctx-m,sulfamilamide resistant genes sul1, sul2, and sul3, a pair of primers of each gene was designed. DNAs of 282 strains isolated from plague natural foci in Qinghai Province were amplified by PCR using every pair of primers. The products were separated using gel electrophoresis, and the results were visualized through a gel imaging system. The susceptibility of 282 Yersina pestis to streptomycin, sulfamethoxazole and ceftriaxone was tested by drug sensitivity test. Results The PCR amplification results of all samples were negative,and strains with streptomycin,sulfamilamide and beta lactam antimicrobial drug resistance genes were not found. Drug sensitivity test showed that 282 strains were highly sensitive to streptomycin,sulfamethoxazole and ceftriaxone sodium.The diameter of bacteriostasis ring>19,17,21 mm, respectively. Conclusions It is a feasible method to use PCR technology to detect the multiple drug resistance genes of Yersinia pestis. Using this method to systematically monitor the resistance gene of Yersinia pestis is an efficient, economical and practical experimental method, which can provide guidance for the treatment of plague disease.

Objective To investigate the clinical significance of abnormally expressed PD-1 on CD 4+CD 2 8+/-T cells in peripheral blood of patients with systemic lupus erythematosus ( SLE ) . Methods Peripheral blood samples were collected form 50 patients with primary SLE and 40 healthy subjects and used to isolated mononuclear cells. Expression of CD4+CD28-, CD4+CD28+, CD4+CD28+PD-1+and CD4+CD28-PD-1+T cells in peripheral blood samples of the two groups were detected by flow cytometry. Clinical data of SLE patients were collected. Based on SLE disease activity index (SLEDAI), SLE patients were classified into two groups: stable group (SLEDAI<10) and active group (SLEDAI≥10). Based on the condition of renal damage, they were also divided into two groups: lupus nephritis group and non-lupus ne-phritis group. Differences in T cell expression were compared among these groups. Statistical analysis was performed to analyze the relationships of different T cell subsets with laboratory and clinical parameters rela-ting to SLE and SLEDAI. Results The percentages of peripheral CD4+CD28-, CD4+CD28+PD-1+and CD4+CD28-PD-1+T cells of active group were higher than those of stable and healthy control groups ( P<0. 05). Moreover, patients with lupus nephritis had higher percentages of these T cell subsets than those without (P<0. 01). SLE patients who were positive for anti-dsDNA or anti-SmRNP antibody, or had de-creased complement C3, thrombocytopenia or decreased lymphocytes had higher percentages peripheral CD4+CD28-T cells than those in the corresponding negative group. SLE patients who were positive for anti-dsDNA or anti-SmRNP antibody, or had decreased complement C3, complement C4 or lymphocytes showed en-hanced expression of peripheral CD4+CD28+PD-1+T cells as compared with those in the corresponding nega-tive group. SLE patients positive for anti-dsDNA antibody, or with decreased complement C3 or lymphocytes or suffering from alopecia had higher percentages of peripheral CD4+CD28-PD-1+T cell than those in the cor-responding negative group. Differences between different groups were statistically significant (P<0. 05). Conclusion Abnormal expression of CD4+CD28-T cells and PD-1 on CD4+CD28-and CD4+CD28+T cells in peripheral blood of patients with SLE has certain correlation with laboratory parameters and clinical indicators.

Objective To investigate the etiology and the epidemiologic features of drug resistance and disinfectant resistance of Yersinia pestis in Hainan,Qinghai Province,in order to provide a scientific basis for prevention and control of the plague in this area.Methods Totally 75 strains were isolated from vary kinds of host in Hainan from 1960 to 2009,and biochemical test,virulence factors evaluation [Fra1 (F1),pesticin Ⅰ (Pst Ⅰ),virulence antigen (VW),pigmentation (Pgm)],plasmid analysis,different region (DFR) genotyping,drug resistance and disinfectant resistance gene test were carried out.Forty-five strains of Yersinia pestis were selected to determine their toxicity in mice,median lethal dose (LD50) was calculated,and LD50 ＜ 1 000 was defined as strongly toxic.Results Sixty of the 75 strains were Qing-Tibet Plateau ecotype,7 strains were Qilian Mountain ecotype,and the remaining 8 were different ecotypes from the plague foci in Qinghai Plateau.Eighty percent (60/75) contained all the four virulence factors;and 97.78％ (44/45) of the strains were velogenic strains;96.00％ (72/75) of the strains contained 3 kinds of plasmids (Mr:6 × 106,45 × 106 and 52 × 106);the DFR strains had 3 genomovars,which were genomovar 8 (65 strains),genomovar 5 (8 strains) and genomovar 21 (2 strains).No strains related to streptomycin,sulfonamides,β-lactam antibiotics and disinfectants had been found in the 75 strains of Yersinia pestis.Conclusions The strains isolated in Hainan have the characteristics of Qinghai-Tibet Plateau plague's pathogen,and they have strong toxicity.In view of high mortality of plague,drug resistance and disinfectant resistance gene test should be put into routine monitoring of the plague.

Objective In order to acquaint with the prevalence of Tibetan sheep plague in this area, we conducted a serum epidemiological investigation of Tibetan sheep plague in Qinghai Province. Methods Indirect hemagglutination assay (IHA) and colloidal gold immunochromatography (GICA) were applied to test serum samples of Tibetan sheep and whole blood samples from jugular vein of Tibetan sheep were collected in 8 Prefectures of Qinghai Province from 2013 to 2016. Results A total of 86 positive Tibetan sheep serum samples with plague F1 antibody were detected by both methods, and the positive rate was 0.68％ (86/12710), the samples collected in Xinghai County Hainan Prefecture had the highest positive rate, which was 5.20％ (27/519). The Haixi Prefecture and Yushu Prefecture were historical epidemic areas, the positive rates were 0.65％(15/2313) and 0.26％(6/2293), respectively. Hainan Prefecture, Guoluo Prefacture and Huangnan Prefecture were newly confirmed epidemic areas, the positive rates were 1.61％ (28/1741), 1.01％ (15/1481), and 1.44％(19/1316), respectively. The antibody titers were 1:20 to 1:5120, the samples collected in Maqin County Guoluo Prefecture had the highest titer, namely 1 :5120. Conclusions In Qinghai Province, Tibetan sheep plague is endemic, and there are outbreaks in some regions. So we have to enhance the Tibetan sheep plague monitoring especially in Marmot plague epidemic area.

Objective:To analyze the pathogenic characteristics of Yersinia pestis in a plague natural foci in Qinghai-Tibet Plateau. Methods:In this study, 1 378 strains of Yersinia pestis isolated from different regions, hosts and vectors in Qinghai-Tibet Plateau from 1954 to 2016 were taken as the research objects. Phenotypic characteristics, plasmid spectrum and genotype of the strains were studied by using conventional techniques and molecular biological techniques. The etiology and geographical distribution of the plague were studied. Results:There were 6 biochemical types of Yersinia pestis in Qinghai-Tibet Plateau, namely Qinghai-Tibet Plateau, Qilian Mountain, Gangdis Mountain, Kunlun Mountain A, Kunlun Mountain B and Chuanqing Plateau. This study found that the Qinghai-Tibet Plateau type strain was not only distributed in north Tibet Plateau, but also distributed in south Tibet, and the distribution of Gangdis Mountain type strain extended to south Tibet. Four virulence factors (capsule antigen, yersinin, virulence antigen and pigmentation factor) were found in 79.97% (1 102/1 378) Yersinia pestis. The results also showed that there were 12 kinds of plasmids carried by Yersinia pestis strains in Qinghai-Tibet Plateau, which constituted 17 kinds of plasmid spectrum. There were 3 kinds of the largest plasmids with taxonomic properties, forming their respective relatively independent distribution areas. The study of different regions (DFR) type showed that 5, 8, 14, 19, 32 and 44 of 1 378 strains were the main genotypes, and the main genome types had obvious geographical distribution. Conclusions:All the tested strains have the characteristics of plague pathogen in Qinghai-Tibet Plateau. The polymorphism of the main hosts, vectors and the ecological landscape of plague geography in the plague foci in Qinghai-Tibet Plateau may lead to the diversity of biochemical characters, plasmid spectrum and geno types of Yersinia pestis.