Objective:To observe multiple locus variable-number tandem repeat analysis (MLVA) typing of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province, and to explore the relationship between the strains and strains previous isolated from Qinghai Province. Methods:Blood samples of Himalayan marmot were collected in Qinghai-Tibet Plateau of Qinghai Province from March 2019 to October 2020. Pathogens were isolated and cultured from Brucella antibody positive samples identified by using the rose bengal test (RBT). Conventional biological methods and molecular biological methods (BCSP31-PCR and AMOS-PCR) were used for strain identification. At the same time, MLVA method was used to genotype the isolated strains, and cluster analysis was used to analyze the genetic relationships between the strains based on the genotype of 70 Brucella isolated from different hosts in Qinghai Province. Results:A total of 1 466 blood samples of Himalayan marmot were collected from Qinghai-Tibet Plateau. Two strains of Brucella were isolated and cultured from 64 RBT-positive samples, named QH2013054 and QH2013062, respectively. They were identified as Brucella ovis biotype Ⅲ by conventional and molecular biological methods. The MLVA genotyping results showed that QH2013054 and QH2013062 were different at the Bru16 locus, indicating different MLVA genotypes. Cluster analysis showed that strain QH2013054 had the same MLVA genotype as 7 strains, among which 6 strains were from 3 farmers and 3 sheep from the same family in Gonghe County, and 1 strain was from a farmer in Menyuan Hui Autonomous County. The strain QH2013062 had the same MLVA genotype as 4 strains, including 3 strains from 3 farmers in Menyuan Hui Autonomous County and 1 strain from a farmer in Tu Autonomous County of Huzhu. Conclusions:The strains of Brucella isolated from Himalayan marmot in Qinghai-Tibet Plateau of Qinghai Province have the same MLVA genotype as some strains of Brucella isolated from humans and sheep in Qinghai Province. It is speculated that the host humans, sheep and Himalayan marmot in Qinghai-Tibet Plateau may have a common source of infection.

Objective:To learn about the epidemic situation and trend of human brucellosis in Menyuan Hui Autonomous County (referred to as Menyuan County) of Qinghai Province, and to provide reference for formulating brucellosis prevention and control measures in Menyuan County.Methods:Data on human brucellosis in Menyuan County reported by the Infectious Disease Reporting Information Management System of China Disease Control and Prevention Information System from 2013 to 2020 were collected and analyzed by descriptive epidemiological analysis (three distribution).Results:A total of 186 cases of brucellosis were confirmed from 2013 to 2020, with an average annual incidence of 14.553/100 000. The annual incidence rate was increasing year by year(χ 2trend = 22.08, P = 0.002). The cases were distributed in 67 villages of 12 towns; cases were more common in the age group of 15-< 65 years old (96.24%, 179/186), and there were more men than women cases (sex ratio was 3.89∶1.00, 148/38). Conclusions:The incidence rate of brucellosis cases in Menyuan County is increasing year by year, and the scope of its impact is constantly expanding. A multi-sectoral joint prevention and control mechanism should be established to strengthen management, increase the prevention and control of brucellosis, and stop the spread of the epidemic to surrounding counties and cities.

Objective:To master the epidemic trend of human brucellosis in Qinghai Province, so as to provide basis for scientific prevention and control of the disease.Methods:In 2019 and 2020, at the national and provincial brucellosis monitoring sites in Qinghai Province, a total of 18 counties (cities and districts, hereinafter referred to as counties), no less than 400 serum samples were sampled every year for brucellosis Rose-Bengal plate agglutination test (RBPT) and serum tube agglutination test (SAT), which would be tested and judged according to the criteria of "Diagnosis for Brucellosis" (WS 269-2019).Results:In 2019, a total of 1 612 people were monitored in national brucellosis monitoring sites, 93 were RBPT positive, 54 were SAT positive, 54 were diagnosed, and the prevalence rate was 3.35% (54/1 612). In 2020, 1 677 people were monitored in national brucellosis monitoring sites, 151 were RBPT positive, 80 were SAT positive, 80 were diagnosed, and the prevalence rate was 4.77% (80/1 677). There were significant differences in RBPT positive rate, SAT positive rate and prevalence rate among national monitoring sites between the two years (χ 2 = 12.52, 4.24, 4.24, P < 0.05). In 2019, a total of 6 043 people were monitored in provincial brucellosis monitoring sites, 128 were RBPT positive, 91 were SAT positive, 87 were diagnosed, and the prevalence rate was 1.44% (87/6 043). In 2020, 5 664 people were monitored, 108 were RBPT positive, 59 were SAT positive, 52 were diagnosed, and the prevalence rate was 0.92% (52/5 664). There was no significant difference in RBPT positive rate among provincial monitoring sites between the two years (χ 2 = 0.66, P = 0.416), and the differences in SAT positive rate and prevalence rate were statistically significant among provincial monitoring sites between the two years (χ 2 = 4.98, 14.57, P < 0.05). Conclusion:In 2019 and 2020, there are human brucellosis in national and provincial brucellosis monitoring sites in Qinghai Province.

Objective:To analyze the causes and epidemiological characteristics of an outbreak of brucellosis in Haidong City Qinghai Province, and provide a scientific basis for prevention and control of brucellosis.Methods:Using the retrospective research method, the epidemic data on brucellosis, field case investigations, serological test results in Haidong City, Qinghai Province in 2019 were collected from the "China Disease Prevention and Control Information System", and the above data were analyzed by descriptive epidemiological analysis.Results:There were 3 confirmed brucellosis patients, who were breeders of the farm; 1 case of latent infection was found in key population, who was villager from the same village as the confirmed patients; 4 people had a history of close contact with infected sheep. A total of 110 sheep serum and 7 bovine serum samples were collected in the animal breeding area, of which 26 were positive, all were sheep serum; 568 sheep serum and 17 bovine serum samples in the same village were negative.Conclusions:The key occupational groups contacted with infected sheep without protection is the main cause of this epidemic. Health education on brucellosis prevention and control should be vigorously carried out to improve the awareness of the people to prevent the disease; the monitoring of animal brucellosis by relevant departments should be strengthened to effectively control the spread of animal brucellosis, and prevent the occurrence of human brucellosis.

Objective:Combined with the actual work requirements of brucellosis in Qinghai Province, to design a management information system for brucellosis patients, in order to realize the information of diagnosis and treatment of brucellosis patients.Methods:Based on the theory of management information system, according to the functional requirements of the actual work and performance parameters, the general framework of management information system was designed, the establishment of the management information system was gradually completed, and the system was improved through the system testing.Results:The system was easy to use and easy to operate, and patient's detailed information, diagnosis and treatment results and treatment plan could be obtained quickly. At the same time, follow-up, trace investigation, and brucellosis prevention and treatment propaganda could be carried out.Conclusions:The established brucellosis management information system can realize the electronic management of patient information, master the disease development and diagnosis and treatment results of brucellosis patients. Through follow-up investigation, the system can increase active screening and early warning of suspected cases, and provide health education to realize early diagnosis and early treatment of the patients.

Objective:To detect cartilage oligomeric matrix protein (COMP) and C-terminal telopeptide of collagen type Ⅱ (CTX-Ⅱ) among brucellosis patients at early period, and provide a better understanding of the osteoarticular changes in this disease at early stage.Methods:Using case-control research method, the early brucellosis patients who visited the Department for Brucellosis Disease Prevention and Control, Qinghai Institute for Endemic Disease Prevention and Control from 2013 to 2016 were selected as the case group; through the principle of random matching, the healthy physical examination population was set as the control group. Serum COMP and CTX-Ⅱ were detected by enzyme-linked immunosorbent assay.Results:A total of 128 patients were investigated, including 61 patients (41 males and 20 females) with early brucellosis, aged (39.41 ± 10.51) years old, and 67 controls (44 males and 23 females), aged (40.31 ± 12.52) years old. The median of serum COMP in the case group was 1.70 μg/L, which was significantly higher than that in the control group (0.79 μg/L). The difference between the two groups was statistically significant ( Z = 3.614, P < 0.05); the median of serum CTX-Ⅱ in the case group was 116.44 pg/ml, and the median of serum CTX-Ⅱ in the control group was 128.48 pg/ml, and the difference was not statistically significant ( Z = 0.044, P > 0.05). Conclusion:The serum COMP levels of brucellosis patients at early period are increased, indicating osteoarticular changes of human brucellosis at early stage.

Objective:The epidemic regularity and trend of human brucellosis in Qinghai Province were analyzed to provide basis for formulating more perfect prevention and control measures.Methods:Using descriptive epidemiological methods, the data of brucellosis reported cases in Qinghai Province from 2005 to 2018 (from the Infectious Disease Report Information Management System of the China Disease Prevention and Control Information System) were collected, and the three distributions (time, region and population distributions) of human brucellosis were statistically described and analyzed.Results:From 2005 to 2018, a total of 418 cases were reported in Qinghai Province, with a incidence of 0.05/100 000, the incidence of different years was statistically different (χ 2 = 11.63, P < 0.05). The incidence of brucellosis remained at a low level from 2005 to 2010, ranged from 0 to 0.05/100 000; and the incidence increased from 2011 to 2018, ranged from 0.27/100 000 to 2.06/100 000. There were reported cases of brucellosis throughout the year, and the onset time was mainly from April to June, accounting for 42.82% (179/418). The 418 cases were distributed in 20 counties (cities, districts), among which the top 5 incidence rates were Menyuan County (15.31%, 64/418), Henan County (13.40%, 56/418), Tianjun County (13.16%, 55/418), Xining City (12.44%, 52/418), and Haiyan County (7.66%, 32/418). In the population distribution, brucellosis cases in the age group from 40 to 49 years old were the most, accounting for 32.78% (137/418), the male to female ratio was 1.58 ∶ 1.00 (256∶162) . And the occupational distribution was dominated by herdsmen, accounting for 57.18% (239/418). Conclusions:The epidemic situation of brucellosis in Qinghai Province is on the rise. It should be strengthened the population monitoring and notification of outbreaks, and health education for all should be implemented.

Objective:To analyze the epidemiological and molecular characteristics of human brucellosis in Qinghai province from 2005 to 2019 and provide basic data for brucellosis prevention and control.Method:The data about human brucellosis in Qinghai from 2005 to 2019 were collected from the information system of China CDC to describe the spatial, population and time distributions of human brucellosis cases in Qinghai. The isolated strains were identified and typed with traditional methods, BCSP31-PCR, AMOS-PCR and multi-locus variablenumber tandem repeat (MLVA-16).Results:A total of 577 human brucellosis cases were reported in Qinghai from 2005 to 2019, the average prevalence rate was 0.07 per 100 000 person, there were statistic differences among different years. The disease occurred all the year around, but mainly during March-October. The 577 cases were distributed in 31 counties (cities/districts) from 6 autonomous prefectures (cities). The prevalence rats of five counties were high, i.e. Menyuan Hui autonomous county (22.88 %, 132/577), Tianjun county (10.57 %, 61/577)、Xining city (10.57 %, 61/577), Henan Mongol Autonomous County (10.51 %, 58/577) and Haiyan county (9.53 %, 55/577). Age of the cases ranged from 8 years to 82 years, and the male to female ratio of the cases was 1.8∶1 (374/203). The prevalence rate in herdsman (47.83 %, 276/577) was highest among different occupational populations. Ten isolates were all Brucella melitensis strains, belonging to biovar 3, and clustering analysis indicated that the 10 strains had 5 genotypes, in which 2 were distinct, the remaining 3 were same. MLVA-16 analysis indicated that the 10 strains had close relationship with 26 B. melitensis strains isolated in Qinghai previously. Conclusions:The prevalence of brucellosis increased in Qinghai in recent years, we should strengthen the population based brucellosis surveillance and reporting. MLVA-16 indicated the gene diversity of the Brucella strains, suggesting that MLVA-16 can be used for genetic diversity analysis and molecular epidemiology survey to improve brucellosis surveillance.

OBJECTIVE： To design and synthesize N-aroyl substituted indoline-3-acetic acid derivatives and evaluate their in vitro hypoglycemic activity. METHODS： Using indoline derivative 2-[5-（benzyloxy）-1-（4-chlorobenzoyl）-2-methyl-1H-inclol-3-yl]acetic acid （GY3） as leading compound， 4-（benzyloxy）phenyl hydrazine hydrochloride and methyl 4-oxopentanoate as raw material， 8 kinds of N-aroyl （3-hydroxybenzoyl， 3-cyanobenzoyl， 4-nitrobenzoyl， 4-methylsulfonylbenzoyl， 4-acetamidobenzoyl， 3-acetylaminobenzoyl， isoniacyl and pyridine-2-formyl） substituted indoline-3-acetic acid derivatives were synthesized via 4 steps reactions： Fischer indole cyclization， reduction， amidation and hydrolyzation. The human hepatoma HepG2 cell lines were used to investigate the glucose consumption activity of the target compounds. RESULTS： Totally 8 various N-aroyl substituted indoline-3-acetic acids were synthesized and their structures were confirmed by mass spectrum（MS）， nuclear magnetic resonance 1H-NMR and 13C spectrum. Under the condition of 1.0 μmol/L， the percentage of glucose- promoting consumption of the synthesized compounds on HepG2 cells was 5.4％-9.1％. 2-[（2R， 3S）-5-benzyloxy-2-methyl-1-（4-methylsulfonyl benzoyl）-2，3-dihydro-indole-3-yl] acetic acid showed the best hypoglycemic activity. The percentage of glucose- promoting consumption was （9.1±1.81）％， which was close to that of positive control metformin [（10.58±1.68）％]， but less potent than that of leading compound GY3[（12.15±0.78）％]. CONCLUSIONS： Different electron-withdrawing substituents are introduced into N-aroyl aromatic rings of dihydroindole compounds， such as cyano， nitro， methyl sulfonyl； hypoglycemic activity decreases in varying degrees and is weaker than halogen substituents.

Objective To understand the changes of serum tumor necrosis factor-α (TNF-α) among adults with Kaschin-Beck disease (KBD) in Qinghai,and to provide scientific data for KBD prevention and control.Methods A case-control study was used,according to "Diagnosis of Kaschin-Beck disease",104 cases of KBD patients [aged (47.61 ± 12.72) years old;50 males,54 females] and 95 cases of internal healthy controls [aged (48.28 ±14.87) years old;35 males,60 females],which were from KBD areas in Xinghai and Guide counties of Hainan Tibetan Autonomous Prefecture,Qinghai Province,and 73 cases of external healthy controls [aged (51.88 ± 13.93)years old;35 males,38 females] from non-KBD region of Xunhua County were included in July 2017.Serum TNF-α was determined via the enzyme linked immunosorbent assay (ELISA) method.Results Totally 272 persons were studied and divided into three groups,there was no statistical significant difference of ages among the three groups (F =2.236,P ＞ 0.05),meanwhile,there was no statistical significant difference of genders among the three groups (x2 =3.135,P ＞ 0.05).The serum TNF-α levels of KBD patients,internal and external healthy controls were (25.14 ± 10.29),(21.84 ± 9.37),and (24.98 ± 8.98) ng/L,respectively,which were statistically significantly different among the three groups (F =3.486,P ＜ 0.05).The serum TNF-α levels in the KBD group and the external healthy control were higher than that in internal healthy control (P ＜ 0.05).There was no significant difference between the KBD group and the external healthy control (P ＞ 0.05).Conclusion There may exist inflammatory changes among KBD patients in KBD areas of Qinghai Province.