ObjectiveTo explore the mechanism of modified Xiehuangsan in treating tic disorders （TD） based on microglial polarization and the Toll-like receptor 4 （TLR4）/myeloid differentiation factor 88 （MyD88）/nuclear factor （NF）-κB pathway. MethodsSeventy-two Sprague-Dawley （SD） rats were randomly assigned into six groups： control， model， tiapride （0.025 g·kg^-1）， and low-， medium-， and high-dose （12， 24， 48 g·kg^-1， respectively） modified Xiehuangsan， with 12 rats in each group. Except the control group， the other groups received intraperitoneal injection of 3，3'-iminodipropionitrile （IDPN） for 7 consecutive days for the modeling of TD. After successful modeling， the control and model groups were given normal saline via gavage， and the other groups were administrated with corresponding drugs by gavage. After 28 days of continuous intervention， rat behaviors were observed， and the modified Xiehuangsan group showing the best anti-TD effect was selected for deciphering the treatment mechanism. Hematoxylin and eosin staining was conducted to observe morphological changes in the rat striatum. Immunohistochemistry was employed to detect the expression of CD16 and CD206 in the striatum. Real-time PCR was employed to measure the mRNA levels of interleukin-1β （IL-1β）， tumor necrosis factor-α （TNF-α）， IL-4， TLR4， MyD88， and NF-κB p65 in the striatum. Western blot was employed to determine the protein levels of ionized calcium-binding adapter molecule 1 （Iba1）， Fc receptor family for immunoglobulin （Ig）G type Ⅲ （CD16）， mannose receptor （CD206）， TLR4， MyD88， and NF-κB p65 in the striatum. ResultsCompared with the control group， the model group showed increased stereotyped behaviors， locomotor activity， total movement distance， and movement speed， shortened resting time （P<0.01）， and noticeable pathological changes in the striatum. Compared with the model group， the tiapride group and modified Xiehuangsan groups exhibited reduced stereotyped behavior， locomotor activity， total movement distance， and movement speed， prolonged resting time （P<0.05， P<0.01）， and alleviated pathological changes in the striatum. Among the modified Xiehuangsan groups， the high-dose group had the best intervention effect and the mildest pathological changes. Therefore， the high-dose group was selected for further research. Compared with the control group， the modeling of TD increased Iba1 and CD16 expression （P<0.05， P<0.01）， up-regulated the mRNA levels of IL-1β and TNF-α （P<0.05， P<0.01）， down-regulated the mRNA level of IL-4 （P<0.05）， up-regulated the mRNA and protein levels of TLR4 and MyD88 （P<0.05， P<0.01）， and up-regulated the protein level of NF-κB p65 （P<0.01）. Compared with the model group， modified Xiehuangsan reduced Iba1 and CD16 expression （P<0.05， P<0.01）， up-regulated the protein level of CD206 （P<0.05， P<0.01）， down-regulated the mRNA levels of IL-1β and TNF-α （P<0.05）， up-regulated the mRNA level of IL-4 （P<0.01）， and down-regulated the mRNA and protein levels of TLR4， MyD88， and NF-κB p65 （P<0.05， P<0.01）. ConclusionModified Xiehuangsan demonstrated a definite therapeutic effect on TD in rats. It may reduce neuroinflammation in TD rats by regulating the polarization of microglia in the striatum via the TLR4/MyD88/NF-κB signaling pathway.

Objective:To investigate the prevalence of adult skeletal fluorosis caused by drinking tea-type endemic fluorosis in Yushu Tibetan Autonomous Prefecture (hereinafter referred to as Yushu Prefecture), Qinghai Province, and provide scientific basis for prevention and control of the disease.Methods:In August 2021, one village was selected as a survey site in six counties (cities) in Yushu Prefecture, including Nangqian, Chindu, Yushu, Zadoi, Qumarlêb, and Zhiduo. Drinking water samples and 10 brick tea samples were collected from each village to determine the fluoride content in water and brick tea; at least 100 permanent residents aged ≥ 25, who had a habit of drinking brick tea and had lived in the local area for more than 5 years, were selected for X-ray imaging to examine the prevalence of adult skeletal fluorosis.Results:A total of 75 samples of residential drinking water were collected, with a fluoride content of (0.21 ± 0.05) mg/L, ranging from 0.11 to 0.34 mg/L; 60 samples of brick tea, with a fluoride content of (626.70 ± 157.27) mg/kg, ranging from 324.00 to 2 102.00 mg/kg. A total of 1 136 adults were examined, and 318 cases of skeletal fluorosis were diagnosed, with a detection rate of 27.99%. Among them, the detection rates of mild, moderate, and severe skeletal fluorosis were 20.95% (238/1 136), 6.07% (69/1 136), and 0.97% (11/1 136), respectively, with mild symptoms being the main. The detection rates of skeletal fluorosis in males and females were 29.09% (121/416) and 27.36% (197/720), respectively, with no statistically significant difference between the gender (χ 2 = 0.39, P = 0.533). Comparison of the skeletal fluorosis in different gender, the differences were statistically significant (χ 2 = 22.31, P < 0.001). The detection rates of skeletal fluorosis in the age groups of 25 - 35, 36 - 45, 46 - 55, 56 - 65, 66 - 75, and ≥76 years old were 6.86% (7/102), 22.37% (51/228), 24.02% (92/383), 37.44% (73/195), 43.48% (70/161), and 37.31% (25/67), respectively. The differences between the groups were statistically significant (χ 2 = 59.84, P < 0.001). Moreover, there was a statistically significant difference in the composition of skeletal fluorosis among different age groups ( H = 37.66, P < 0.001). The Spearman correlation analysis results showed that the severity of adult skeletal fluorosis was positively correlated with age ( r = 0.34, P < 0.001). Conclusions:There is a certain degree of prevalence of adult skeletal fluorosis in Yushu Prefecture. And as age increases, the condition of skeletal fluorosis becomes more severe.

Objective:To investigate the common viral pathogen spectrum of acute respiratory infection (ARI) cases in Changshu city.Methods:Nasopharyngeal swab samples from outpatients and inpatients in five sentinel hospitals in Changshu city from January 2022 to October 2023 were collected. Real-time fluorescent quantitative polymerase chain reaction was used to detect the nucleic acids of influenza virus (Flu) and respiratory syncytial virus (RSV), adenovirus (AdV), human rhinovirus (HRV), human parainfluenza virus (HPIV), enterovirus (EV), human coronavirus (HCoV), human metapneumovirus (HMPV), and human bocavirus (HBoV).Results:Of the 1 936 ARI cases, 18.1% (350/1 936) were tested positive for viral nucleic acid. Flu had the highest detection rate (7.1%), followed by HBoV (3.7%), AdV (1.9%) and RSV (1.6%). The virus detection rate was significantly different among different age groups and seasons. In March 2023, the total virus detection rate was the highest (47.6%), mainly Flu. Flu showed unimodal prevalence in spring and winter. The prevalence trend of HBoV and HCoV was consistent, and the detection rate of HBoV (5.43%) was higher than that of HCoV (2.45%), both of which peaked in summer. RSV prevalence peaks in autumn (4.5%). The detection rate of children aged 0-5 years was the highest, reaching 33.3%.Conclusions:The main ARI pathogens in Changshu city from 2022 to 2023 were Flu, HBoV, AdV and RSV.

Objective:To determine incidence of seasonal influenza virus infection in the community and to analyze the factors influencing seasonal influenza virus infection.Methods:This study recruited residents aged 6-59 years to build a cohort in 15 villages/streets in Macheng city in November 2019. Meanwhile, a cross-sectional baseline survey was conducted immediately to collect sera, information on demographics and child protection knowledge, behaviors, as well as attitudes using a questionnaire from the participants enrolled in the cohort (i.e., before the influenza epidemic season). In July 2020, a cross-sectional follow-up survey was conducted to collect sera once again (i.e., after the influenza season). Paired sera from the two cross-sectional surveys were tested for influenza virus-specific antibodies by hemagglutination inhibition (HI) test or micro-neutralization (MN) test using a circulating representative strain of each subtype/lineage of influenza virus as the test antigen. The infections with influenza virus subtype/lineage was confirmed if there was a four-fold or more increase in titers of antibodies against circulating representative strain of the subtype/lineage of influenza virus. Factors influencing infection with influenza A (H3N2) and B/Victoria viruses were analyzed using univariable and multivariable logistic regression.Results:In November 2019, 800 study participants were enrolled in the cohort, including 340 children aged 6-17 years and 460 adults aged 18-59 years; 605 study participants (including 224 children and 381 adults) were followed up in July 2020 and their paired sera were obtained before and after the influenza season. 25.3% (153/605) of the participants were confirmed to be infected with at least one subtype/lineage of seasonal influenza virus by HI and MN tests. The overall incidence of influenza viruses of all subtypes/lineages in children was 44.2% (95% CI: 37.6%-50.8%) which was significantly higher than the incidence of 14.1% in adults (95% CI: 10.7%-17.7%). Children had the highest incidence of influenza A (H3N2) virus infection, followed by B/Victoria. MN or HI antibody titers in A (H3N2)[ OR=0.88 (95% CI: 0.84-0.93)] and B/Victoria[ OR=0.97 (95% CI: 0.95-0.99)] before the influenza season were significantly associated with whether children were infected with that subtype/lineage of influenza virus. Conclusions:The residents aged 6-59 years in Macheng city had a substantial incidence of seasonal influenza virus infection during the influenza season from winter 2019 to spring 2020. Notably, almost half of children aged 6-17 years have been infected with seasonal influenza virus. Higher titers of HI/MN antibodies against seasonal influenza virus before the influenza season would be likely to reduce the risk of infection with influenza A (H3N2) and B/Victoria.

Objective:To construct a prediction model for peripherally inserted central catheter-related upper extremity deep vein thrombosis (PICC-UEDVT) in patients with traumatic brain injury (TBI) and validate its effectiveness.Methods:A case-control study was conducted on the clinical data of 222 TBI patients admitted to Xiangya Hospital of Central South University from January 2019 to December 2021, including 171 males and 51 females, aged 18-86 years [54.5(46.0, 65.0)years]. Glasgow coma scale (GCS) motor score was 4.0(3.0, 5.0)points on the day of catheterization. A total of 82 patients (36.9%) had PICC-UEDVT. The patients were randomly divided with a ratio of 7∶3 into training set ( n=156, including 58 with PICC-UEDVT) and validation set ( n=66, including 24 with PICC-UEDVT) using R programming language. The baseline data of general information, intravenous medication, catheterization, and laboratory indices were compared between the training set and the validation set. Lasso regression analysis was employed to identify those variables, with the diagnosis of PICC-UEDVT as the outcome variable. Variables with non-zero regression coefficients were included in a multifactorial Logistic regression model and independent variables were selected based on the Akaike Information Criterion (AIC) of R programming language. The regression equation was constructed, based on which, the predictive nomogram model was constructed for PICC-UEDVT in TBI patients. Receiver operating characteristic (ROC) curves for the training set and validation set were plotted and the discriminability of the model was assessed. The calibration of the model was evaluated using the Hosmer-Lemeshow (H-L) goodness-of-fit test and calibration curves and the clinical practicality of the model was assessed with decision curve analysis (DCA). Results:The baseline analysis of both the training set and the validation set demonstrated a well-balanced sample distribution. Through Lasso regression analysis, 5 prediction variables were identified: GCS motor score on the day of catheterization, Caprini score on the day of catheterization, use of glucocorticoids, tip position of the catheter, and D-dimer (D-D) level before catheterization. The multivariate Logistic regression analysis revealed that the Caprini score on the day of catheterization ( OR=1.20, 95% CI 1.08, 1.33), use of glucocorticoids ( OR=3.13, 95% CI 0.99, 10.46), and D-D level before catheterization ( OR=1.16, 95% CI 1.07, 1.33) were independent risk factors for PICC-UEDVT in TBI patients. The regression equation was developed as: Logit [ P/(1- P)]=-2.56+0.18×"Caprini score on the day of catheterization"+1.14×"use of glucocorticoids"+0.15×"D-D level before catheterization". In the prediction model which was constructed based on the equation, the AUC values for the training set and validation set were 0.73 (95% CI 0.65, 0.81) and 0.77 (95% CI 0.65, 0.87) respectively. The H-L goodness-of-fit test indicated χ2=3.28, P=0.950 for the training set and χ2=13.05, P=0.160 for the validation set. Calibration curves for both sets demonstrated alignment between the actual and predicted probabilities of PICC-UEDVT in TBI patients. DCA results showed that the net benefit rate of patients was optimal when the threshold probability ranged from 15% to 72% for the training set and from 10% to 81% for the validation set. Conclusion:The prediction model based on the Caprini score on the day of catheterization, use of glucocorticoids, and D-D level before catheterization demonstrates good predictive accuracy, calibration and clinical practicality in predicting PICC-UEDVT in TBI patients.

Objective:To learn about the detection quality and external quality control assessment of fluoride and arsenic in laboratories at all levels in Qinghai Province.Methods:The Z-score method was used to analyze and evaluate the evaluation results of 1 provincial, 8 municipal and 43 county level laboratories of disease prevention and control institutions participating in the external quality control assessment of water fluoride and brick tea fluoride in Qinghai Province in 2021, as well as 1 provincial, 1 municipal and 2 county level laboratories of disease prevention and control institutions participating in the external quality control assessment of water arsenic and urine arsenic. The feedback rate and qualification rate of external quality control of each assessment laboratory were calculated.Results:In 2021, the feedback rate of external quality control of water fluoride, brick tea fluoride, water arsenic and urine arsenic in provincial and municipal level laboratories of Qinghai Province were 100.00%; except that the qualified rate of water fluoride was 7/9, the qualified rate of external quality control of other projects was 100.00%. The feedback rate of external quality control of water fluoride, brick tea fluoride, water arsenic and urine arsenic in county level laboratories was 100.00%; except that the qualified rate of water fluoride was 86.05% (37/43), the qualified rate of external quality control of other projects was 100.00%. In the specific assessment results of the laboratory, the assessment results of water fluoride sample FS20210101 from 1 provincial, 1 municipal and 2 county level laboratories, and FS20210102 from 1 county level laboratory were suspicious; the assessment results of water fluoride sample FS20210101 from 3 county level laboratories were not satisfactory; the assessment results of fluoride and arsenic sample in other laboratories were satisfactory.Conclusions:The qualified rate of external quality control of fluoride and arsenic in laboratories at all levels in Qinghai Province is relatively high, but some county level laboratories are still dissatisfied with the assessment results of water fluoride. Therefore, it is necessary to strengthen the detection level of water fluoride in laboratories.

Objective:To observe the changes of serum C-terminal peptide of type Ⅰ collagen (CTX-1) and N-terminal lengthening peptide of type Ⅰ collagen (P1NP) in adult patients with skeletal fluorosis in the tea-drinking-borne endemic fluorosis area in Qinghai Province, and to find sensitive indicators for diagnosis of skeletal fluorosis.Methods:From April to August 2019, a case-control study was carried out in tea-drinking-borne endemic fluorosis area in Zhiduo County, Yushu Tibetan Autonomous Prefecture, and Gangcha County, Haibei Tibetan Autonomous Prefecture of Qinghai Province. According to the Diagnostic Standard for Endemic Skeletal Fluorosis (WS/T 192-2008), the clinical diagnosis and X-ray examination of skeletal fluorosis were carried out for permanent residents ≥25 years old and living for more than 10 years in the area, combined with face-to-face inquiry and investigation of past disease history, lifestyle and clinical manifestations. The patients with skeletal fluorosis and healthy people were selected as skeletal fluorosis group and control group, respectively. Randomized urine samples and fasting venous blood from the two groups were collected. The content of fluoride in urine was determined by ion selective electrode method, and the contents of CTX-1 and P1NP in serum were determined by enzyme-linked immunosorbent assay (ELISA).Results:A total of 127 people in the disease area were investigated, including 63 cases in skeletal fluorosis group and 64 cases in control group. There was no statistically significant difference in age and sex ratio between the two groups ( t = 0.42, χ 2 = 0.07, P > 0.05). The X-ray examination results showed that the patients with skeletal fluorosis were mainly mild, accounting for 71.43% (45/63); X-ray changes were mainly ossification of interosseous membrane and tendon. The urinary fluoride in control group and skeletal fluorosis group was 1.62 (1.12, 1.95) and 3.22 (2.38, 4.89) mg/L, respectively, with statistically significant difference between the two groups ( Z = 7.07, P < 0.001). The difference of serum CTX-1 and P1NP contents between the two groups was statistically significant ( Z = 2.00, 4.89, P < 0.05). Conclusions:The levels of serum CTX-1 and P1NP in patients with skeletal fluorosis are higher than those in healthy people. Serum CTX-1 and P1NP may be used as sensitive indicators for diagnosis of skeletal fluorosis.

Abstract OBJECTIVE To investigate the specific binding of nucleic acid aptamers(k-α2ct) with plasminogen Kringle 5(K5) on the function of K5 in inhibiting proliferation and migration of vascular endothelial cells and promoting their apoptosis. METHODS The cloning and expression of recombinant K5 protein were performed by using a prokaryotic system, and the isolation and purification of the expressed K5 protein were performed by affinity chromatography. The affinity and specificity of K-a2ct and K5 were verified using isothermal titration calorimetry(ITC) and enzyme-linked oligonucleotide adsorption assay(ELONA). The effect of K-a2ct on the function of K5 in inhibiting the proliferation and migration of human umbilical vein endothelial cells(HUVEC) was investigated by CCK-8 and cell scratch assay. The apoptotic morphology of HUVEC cells stained with Hoechst 33342 was observed by laser confocal microscopy, and the effect of K-a2ct on the apoptosis- promoting function of K5 in HUVEC cells was also examined by Annexin V/PI double-stained flow cytometry. RESULTS Recombinant protein K5 was efficiently expressed in Escherichia coli and purified by affinity chromatography, identified as having a relative molecular weight of 12 kDa and a concentration of 0.32 mg·mL-1. ITC and ELONA results demonstrated that K-a2ct had a strong affinity and good selectivity for K5, showing the affinity specificity of a typical nucleic acid aptamer. CCK-8 and cell scratching assays showed that K-a2ct could inhibit the anti-proliferation and anti-migration effects of K5 on HUVEC cells in a dose-dependent manner. The laser confocal and flow cytometry results showed that K-a2ct inhibited the apoptosis-promoting function of K5 on HUVEC cells, mainly affecting late apoptosis of HUVEC cells effected by K5 but having little effect on early apoptosis. CONCLUSION The nucleic acid aptamers K-a2ct binds to K5 with high affinity and specificity, and inhibits its anti angiogenic function in a dose-dependent manner. It has great potential in targeting the regulation of K5 concentration and function in vivo and promoting angiogenesis.

Objective:To evaluate the water fluoride detection ability of county (city, district) level (referred to as the county-level) laboratories in Qinghai Province.Methods:During the "13th Five-Year Plan" period (2016 - 2020), 4, 4, 4, 43, 43 county-level laboratories in Qinghai Province were organized to participate in the national water fluoride external quality control assessment, and the assessment results were evaluated by Z-ratio scoring method.Results:The response rate of county-level laboratories was 86.05% (37/43) in 2019, and 100.00% in other years. The qualified rate of county-level laboratories was 100.00% from 2016 to 2018; in 2019 and 2020, the assessment was fully covered, and the qualified rate was 81.40% (35/43) and 95.35% (41/43), respectively. Compared with 2019, the response rate and qualified rate in 2020 increased significantly, and the differences were statistically significant (χ 2 = 6.450, 4.074, P < 0.05). In the past 5 years, two assessment samples│Z│ < 1 in each laboratory were the most common, but with two assessment samples 2≤│Z│ < 3 in some qualified laboratories. Conclusion:The consistency of water fluoride determination in Qinghai Province is not very ideal, and the detection ability of county-level laboratories still needs to be strengthened.

Objective:To understand the detection quality and external quality control assessment of fluorine and arsenic laboratories at all levels in Qinghai Province.Methods:Z-ratio scoring method was used to analyze and evaluate the results of provincial, municipal and county-level laboratories participating in the national fluorine and arsenic external quality control assessment from 2014 to 2020 in Qinghai Province, │Z│≤ 2 was qualified, 2 <│Z│ < 3 was basically qualified and│Z│≥ 3 was unqualified. At the same time, the feedback rate and pass rate of the participating laboratories were calculated.Results:From 2014 to 2020, the external quality control assessment results of water arsenic, urinary arsenic and brick tea fluorine in Qinghai Provincial laboratory were all qualified; except for the external quality control assessment of water fluorine in 2018 that was basically qualified, the results of other years were all qualified. Municipal and county-level laboratories did not give feedback the external quality control assessment results of water arsenic and urinary arsenic in 2017, and the feedback rate and pass rate in other years were 100.0%; except for the external quality control feedback rate and pass rate of water fluorine and brick tea fluorine in 2014 that were all 100.0%, there were laboratories that did not give feedback or failed the assessment in other years.Conclusion:The testing ability of provincial laboratory in Qinghai Province meets the standard, and the testing ability of municipal and county-level laboratories still need to be improved continuously.