Objective: To evaluate the effectiveness for the construction of "mosquito-free village" in Xuejia Village, Pujiang County, Zhejiang Province, so as to provide the guidance for the construction of "mosquito-free village" in other rural areas. Methods: Density of adult mosquitoes in Xuejia Village was investigated using light trap method and density of larval mosquitoes was investigated using larval straw method from April to November each year. Totally 30 households of villagers were randomly selected, and their awareness rates of mosquito control knowledge, mosquito control behavior forming rates and satisfaction rates were surveyed through questionnaires. Investment during construction (from August 2016 to December 2018) and maintenance period (from 2019 to 2023) of "mosquito-free village" were investigated through data review and interviews. Long-term effects of "mosquito-free village" construction in Xuejia Village were evaluated in terms of mosquito density from 2016 to 2023, effectiveness of health education for villagers, satisfaction and investment. Results: Compared with the year 2016, the densities of adult and larval mosquitoes in Xuejia Village were significantly decreased from 2017 to 2023. The average monthly density of adult mosquitoes in 2023 decreased by 98.34%, and the average monthly 100 household index of larval mosquitoes decreased by 98.45% compared to 2016. The average monthly density of adult mosquitoes from 2019 to 2023 was less than or equal to one mosquito per light trap in a night, and the average monthly 100 household index of larval mosquitoes was less than or equal to five places per 100 households. The awareness rate of mosquito control knowledge was 93.33%, the behavior forming rate was 86.67%, and the satisfaction rate was 90.00%. By December 2023, the total investment during construction and maintenance period was 450 thousand Yuan, with an average annual investment of 60.7 thousand Yuan and average annual investment of 206.61 Yuan per household. The average annual investment during maintenance period was 36.2 thousand Yuan, and the average annual investment per household was 109.70 Yuan. Conclusion The mosquito density, effectiveness of health education for villagers and satisfaction of "mosquito-free village" in Xuejia Village all meet the evaluation criteria of "mosquito-free village", and the investment is reasonable, making it suitable for promotion to other rural areas.

Objective:The epidemiological and host animal pathogen data of leptospirosis in the population of Zhejiang Province in 2023 were analyzed, providing scientific basis for formulating prevention and control strategies of leptospirosis.Methods:The data of human leptospirosis in the population were collected from the China Information System for Disease Control and Prevention, and analyzed using descriptive epidemiological methods. The data on isolation, culture, and nucleic acid testing of Leptospira pathogens from mouse kidneys, frog kidneys, pig kidneys, and duck kidneys as well as duck serum antibody data were collected from Zhejiang Provincial Center for Disease Control and Prevention "Leptospirosis Surveillance Project of Zhejiang Province". The carrying and changing status of Leptospira epidemic microbiota in populations and host animals were analyzed. Results:In 2023, a total of 83 cases of leptospirosis were reported in Zhejiang Province, with a incidence rate of 0.126 2/ 100 000, aged (62.66 ± 11.31) years, including 68 males and 15 females. Leptospirosis cases were reported in 11 cities, mainly concentrated in the southern cities of Wenzhou City, Lishui City and Taizhou City(a total of 68 cases), accounting for 81.93% of the total number of cases. August to October were high incidence months for leptospirosis (a total of 70 cases), accounting for 84.34% of the total number cases. The male to female ratio of patients was 4.53 ∶ 1.00, and all were adults ≥20 years old, the middle and old people aged 45 - 79 years were the high-risk population (a total of 77 cases), accounting for 92.77% of the total number of cases. The patient's occupation was mainly farmers, with a total of 54 cases, accounting for 65.06% of the total number cases. The shortest time from onset to initial diagnosis for patients with leptospirosis was 0 day, and the longest was 13 days. The shortest time from initial diagnosis to confirmed diagnosis was 0 day, and the longest was 16 days. The 72.29% of the leptospirosis cases (60 cases) had a history of field labor or suspected contact with contaminated water within one month before the onset of the disease, and 18.07% of the leptospirosis cases (15 cases) had a history of contact with animals such as mice, frogs, pigs, cows, dogs, fish or ducks, or their excreta within one month before the onset of the disease. The average nucleic acid positive rate of host animals with leptospirosis was 5.92% (31/524) in mouse kidney, 6.74% (36/534) in frog kidney, and 0.66% (1/151) in pig kidney. The isolation and culture of leptospirosis from duck kidney, nucleic acid detection, and antibody detection in duck blood were all negative. The leptospirosis bacteria detected in human population were serogroup Icterohaemorrhagiae (3 samples) and Hebdomadis (4 samples), and the bacteria group detected in host animals was serogroup Icterohaemorrhagiae (3 samples). Conclusions:The outbreak of leptospirosis in Zhejiang Province mainly occurs in the summer and autumn, with the affected areas mainly concentrated in the southern region of Zhejiang Province. The affected population is mainly middle-aged and elderly males, and the population carrying Leptospira is still mainly composed of the serogroup Icterohaemorrhagiae and the Hebdomadis, with the host animal being the serogroup Icterohaemorrhagiae.

Traditionally, Tripterygium hypoglaucum (Levl.) Hutch (THH) are widely used in Chinese folk to treat rheumatoid arthritis (RA). This study aimed to investigate whether the anti-RA effect of THH is related with the gut microbiota. The main components of prepared THH extract were identified by HPLC-MS. C57BL/6 mice with adjuvant-induced arthritis (AIA) were treated with THH extract by gavage for one month. THH extract significantly alleviated swollen ankle, joint cavity exudation, and articular cartilage destruction in AIA mice. The mRNA and protein levels of inflammatory mediators in muscles and plasma indicated that THH extract attenuated inflammatory responses in the joint by blocking TLR4/MyD88/MAPK signaling pathways. THH extract remarkably restored the dysbiosis of the gut microbiota in AIA mice, featuring the increases of Bifidobacterium, Akkermansia, and Lactobacillus and the decreases of Butyricimonas, Parabacteroides, and Anaeroplasma. Furthermore, the altered bacteria were closely correlated with physiological indices and drove metabolic changes of the intestinal microbiota. In addition, antibiotic-induced pseudo germ-free mice were employed to verify the role of the intestinal flora. Strikingly, THH treatment failed to ameliorate the arthritis symptoms and signaling pathways in pseudo germ-free mice, which validates the indispensable role of the intestinal flora. For the first time, we demonstrated that THH extract protects joint inflammation by manipulating the intestinal flora and regulating the TLR4/MyD88/MAPK signaling pathway. Therefore, THH extract may serve as a microbial modulator to recover RA in clincial practice.ver RA in clincial practice.
Mice ; Animals ; Gastrointestinal Microbiome ; Tripterygium ; Myeloid Differentiation Factor 88/genetics* ; Toll-Like Receptor 4/genetics* ; Mice, Inbred C57BL ; Arthritis, Experimental/drug therapy*

Esophageal squamous cell carcinoma (ESCC) is one of the leading causes of cancer death worldwide. It is urgent to develop new drugs to improve the prognosis of ESCC patients. Here, we found benzydamine, a locally acting non-steroidal anti-inflammatory drug, had potent cytotoxic effect on ESCC cells. Benzydamine could suppress ESCC proliferation in vivo and in vitro. In terms of mechanism, CDK2 was identified as a target of benzydamine by molecular docking, pull-down assay and in vitro kinase assay. Specifically, benzydamine inhibited the growth of ESCC cells by inhibiting CDK2 activity and affecting downstream phosphorylation of MCM2, c-Myc and Rb, resulting in cell cycle arrest. Our study illustrates that benzydamine inhibits the growth of ESCC cells by downregulating the CDK2 pathway.
Humans ; Benzydamine ; Esophageal Neoplasms/drug therapy* ; Esophageal Squamous Cell Carcinoma/drug therapy* ; Molecular Docking Simulation ; Phosphorylation ; Cell Proliferation ; Cell Line, Tumor ; Apoptosis ; Cyclin-Dependent Kinase 2

Objective:To investigate the epidemic characteristics and current situation of animal plague in Zhejiang Province, in order to provide data support for prevention and control of the plague in the whole province.Methods:A retrospective study was carried out to collect surveillance data of animal plague in Zhejiang Province from 2006 to 2020 from the "Pubonic Plague Control Management Information System". Descriptive epidemiological methods were used to analyze the density of domestic and wild rodents, the distribution of rodent species and flea species, and the serological and pathogenic test results of host animals.Results:From 2006 to 2020, the annual average density of domestic rodent in Zhejiang Province was 3.99%, which was lower than that in 2006, 2007, 2008, 2009, 2019 and 2020. The annual average density of wild rodents was 4.52%, which was lower than that in 2013, 2016, 2017, 2018, 2019 and 2020. From 2006 to 2020, a total of 173 432 rodents were captured, belonging to 3 orders, 5 families, 12 genera and 17 species; in the composition of rodent species, Rattus norvegicus was the most, accounting for 32.15% (55 765/173 432). A total of 107 736 rodents were examined, the number of flea-infected rodent was 3 885, and the flea-infested rate was 3.61%; the total number of flea collected was 9 039 and the total flea index was 0.083 9. A total of 172 235 serological samples were tested at various monitoring sites in Zhejiang Province, and 10 positive samples were detected, there were 7 copies of Apodemus agrarianus, 2 copies of Smelly Shrew and 1 copy of Rattus norvegicus, with titers ranging from 1 ∶ 40 - 1 ∶ 320; a total of 163 618 copies of rodent liver and spleen organs were cultured, and no Yersinia pestis was isolated. Conclusions:Positive host animals have been detected in Zhejiang Province for many years. It is necessary to further standardize plague surveillance and make emergency preparations to prevent relapse and import of the epidemics.

Objective:To investigate the risk factors of bone cement leakage after percutaneous vertebroplasty (PVP) for osteoporotic vertebral compression fracture (OVCF).Methods:A multi-center, large-sample, case-control study was carried out to analyze the clinical data of 2 273 OVCF patients (2 689 vertebrae) undergone PVP at four hospitals between May 2018 and October 2021, including 994 males and 1 279 females, with the age of 52-91 years [(69.1±3.1)years]. Of all, 581 patients (604 vertebrae) were allocated to leakage group and 1 692 patients (2 085 vertebrae) to no leakage group according to the occurrence of bone cement leakage. The gender, age, fracture sites, vertebral compression degree, endplate integrity of fractured vertebrae, surgical segments, surgical approaches and bone cement injection volume were recorded. Univariate analysis was used to investigate the correlation between those indicators with bone cement leakage. Multivariate Logistic regression analysis was used to identify the independent risk factors for bone cement leakage.Results:Univariate analysis showed that gender, age, fracture sites, vertebral compression degree, bone cement injection volume were related to bone cement leakage after PVP ( P<0.05 or 0.01), but no correlation was found in the endplate integrity of fractured vertebrae, surgical segments and surgical approaches (all P>0.05). Multivariate Logistic regression analysis showed that fracture sites ( OR=1.68, 95% CI 1.11-2.55, P<0.05), vertebral compression degree more than 40% ( OR=1.98, 95% CI 1.29-3.02, P<0.01), bone cement injection volume greater than or equal to 5.5 ml ( OR=1.55, 95% CI 1.07-2.26, P<0.05) were significantly associated with bone cement leakage after PVP. Conclusion:Thoracic vertebral fracture, vertebral compression degree more than 40% and bone cement injection volume greater than or equal to 5.5 ml are independent risk factors for bone cement leakage after PVP in OVCF.

Objective:To investigate the characteristics and evolution of mild motor symptoms (MMS) in patients with prodromal Parkinson′s disease (pPD).Methods:Based on the pPD cohort screened by Parkinson′s Disease Prodromal Clinical Assessment Scale in Nanjing community from July 2018 to December 2020, the clinical data of 30 patients with pPD who completed the baseline assessment and were followed up for at least 1 year were analyzed. According to the Unified Parkinson Diease Rating Scale Ⅲ (UPDRS-Ⅲ) score, the patients were divided into MMS group (UPDRS-Ⅲ score>3) and non-MMS group (NMMS group, UPDRS-Ⅲ score≤3). The differences and evolution characteristics of clinical characteristics between the 2 groups were compared. Multivariate linear regression was used to analyze the risk factors of motor symptom progression in pPD patients.Results:Among the 30 patients with pPD, 7 of 23 patients in the MMS group were converted to PD at the end of follow-up, 1 of 7 patients in the NMMS group were converted to PD at the end of follow-up. The UPDRS-Ⅲ score [10.00 (7.00, 17.00)], Montreal Cognitive Assessment Scale (MoCA) score [25.50 (24.75, 28.00)] and the Hamilton Anxiety Scale (HAMA) score [9.00 (5.00, 13.00)] at the end of follow-up of pPD patients were significantly higher than those at baseline [7.00 (4.00, 12.00), 24.00 (22.75, 25.25) and 8.00 (2.00, 11.00)], and the differences were statistically significant ( Z=-3.505, P<0.001; Z=-2.956, P=0.003; Z=-2.427, P=0.015).Subgroup analysis showed that UPDRS-Ⅲ score [11.00 (7.00, 18.00)], MoCA score [25.00 (24.00, 27.00)] and HAMA score [ 9.00 (6.00, 15.00)] at the end of follow-up in the MMS group were higher than those at baseline [8.00 (6.00, 12.00), 24.00 (22.00, 25.00) and 9.00 (3.00, 11.00)], and the difference was statistically significant (Z=-2.768, P=0.006; Z=-2.457, P=0.014; Z=-2.250, P=0.024). The Non-Motor Symptoms Questionnaire score at the end of follow-up in the MMS group (8.96±5.20) was significantly lower than that in the baseline (11.04±4.41), and the difference was statistically significant ( t=2.441, P=0.023).There was no significant difference in Mini-Mental State Examination (MMSE), Hamilton Depression Scale (HAMD), Rapid Eyes Movement Sleep Behavior Disorder Questionnaire-Hong Kong (RBDQ-HK) and Sniffin′ sticks olfactory test score at the end of follow-up in the MMS group. Only UPDRS-Ⅲ score in the NMMS group was increased at the end of follow-up [7.00 (5.00, 8.00)] compared with the baseline [4.00 (1.00, 4.00)], and the difference was statistically significant ( Z=-2.375, P=0.018). There was no significant difference in MoCA, MMSE, HAMA, HAMD, RBDQ-HK, and Sniffin′ sticks olfactory test score between the NMMS group and the baseline at the end of follow-up. Conclusion:The clinical conversion rate of pPD patients with MMS is high,and screening of this population should be paid attention.

Objective:To analyze the epidemiological characteristics and spatiotemporal distribution of hemorrhagic fever with renal syndrome (HFRS) in Zhejiang province from 2005 to 2020, and provide scientific information for the precise prevention and control of HFRS.Methods:Data on HFRS cases in Zhejiang province during 2005-2020 were collected from the China National Notifiable Infectious Disease Reporting Information System (NNDS) for a descriptive analysis, and software ArcGIS 10.2 was used for global autocorrelation and local autocorrelation analyses. Spatiotemporal clusters were scanned with SaTScan 9.4.4 and visualized with ArcGIS 10.2.Results:A total of 7 724 HFRS cases were reported in Zhejiang province from 2005 to 2020, including 25 deaths. There were two incidence peaks each year, in late spring and early summer (May-June) and in winter (November-January). The top three areas with high cumulative cases were Ningbo (1 875, 24.27%), Taizhou (1 642, 21.25%), and Shaoxing (1 123, 14.54%). Among the reported cases, with a male to female ratio of 2.73∶1(5 656∶2 068). The majority of HFRS cases were middle-aged and elderly people, with cases aged 41-70 years accounting for 60.95%. Most HFRS cases were farmers, accounting for 69.89% (5 398/7 724). The spatial distribution of HFRS in most years was correlated. SaTScan was used for retrospective spatiotemporal scanning and three clusters were detected: the first type clusters were in 21 counties in eastern Zhejiang province and central Zhejiang province, among which 4 were in Ningbo, Shaoxing and Jinhua, 8 were in Taizhou, and 1 was in Lishui ( RR=13.69, LLR=5 522.60, P<0.001); the second type clusters were in Longquan and Qingyuan counties ( RR=31.20, LLR=1 232.46, P<0.001); the third types of clusters were in Changxing and Anji counties of Huzhou in northern Zhejiang province ( RR=3.42, LLR=23.93, P<0.001). Conclusions:HFRS mainly occurred in middle-aged,elderly and male farmers in Zhejiang province. The incidence was high in late spring, early summer and winter in eastern Zhejiang province. Precise prevention and control measures are needed for populations at high risk before the epidemic season.

Objective:To design the fourth-generation chimeric antigen receptor-T (CAR-T) cells that secrete interleukin-7 (IL7) and chemokine C legend 19 (CCL19) on the basis of the second-generation CAR, and to analyze and compare the differences in proliferation, chemotaxis, tumor cell clearance and persistence in the microenvironment of multiple myeloma (MM) between them.Methods:The fourth-generation CAR vector plasmid was constructed by using 2A self-cleaving peptide technology. The third-generation lentiviral packaging system was used to prepare high-titer lentivirus. Flow cytometry was used to monitor the transduction efficiency of lentivirus and the subtype changes of CAR-T cells. The enzyme-linked immunosorbent assay (ELISA) was used to quantify the IL7 and CCL19 secreted by CAR-T cells.The calculation of absolute number of CAR-T cells during culture was used to analysis cell proliferation activity. Transwell migration assay was used to verify the chemotactic ability of CAR-T cells. The specific killing activity of CAR-T cells was detected by using the luciferase bioluminescence method. The NOD-Prkdcem26Cd52Il2rgem26Cd22/Nju (NOD) mouse xenograft model was used to verify the anti-myeloma activity and safety of CAR-T cells in vivo.Results:Flow cytometry results showed that the stable CAR expression rates of the second-generation anti-CS1 CAR-T and fourth-generation anti-CS1-IL7-CCL19 CAR-T cells were (91.50±0.29)% and (46.7±0.12)%, respectively. CAR-T cells were successfully constructed. Subtype analysis demonstrated that the ratio of stem memory T cell (TSCM) in anti-CS1-IL7-CCL19 CAR-T cells was (67.58±0.59)%, which was significantly higher than (50.74 ± 1.01)% of anti-CS1 CAR-T ( P=0.000 1), with more strong immune memory function and better durability. Anti-CS1-IL7-CCL19 CAR-T cells can continuously secrete IL7 and CCL19 compared to MOCK-T and anti-CS1 CAR-T ( P<0.000 1). The number of anti-CS1-IL7-CCL19 CAR-T cells reached (22.77±0.79)×10 6 on the 9th day after lentivirus transduction, which was significantly higher than (9.40±0.79)×10 6 of anti-CS1 CAR-T cells ( P=0.000 1), with stronger proliferation ability. The number of chemotaxis cells of anti-CS1-IL7-CCL19 CAR-T cells to reactive T cells was (109.0±4.04), which was significantly higher than (9.33±1.20) of MOCK-T ( P<0.000 1) and (7.33±0.88) of anti-CS1 CAR-T ( P<0.000 1), with stronger chemotactic ability. The specific killing activity showed that both anti-CS1-IL7-CCL19 CAR-T and anti-CS1 CAR-T cells had specific killing efficacies when compared with the MOCK-T cells ( P<0.000 1). Animal experiment indicated that anti-CS1-IL7-CCL19 CAR-T cells significantly reduced the tumor burden ( P<0.000 1) and extended the overall survival time ( P=0.006 1) of tumor-bearing mice. Conclusions:The anti-CS1-IL7-CCL19 CAR-T cells designed in this study show stronger proliferative activity, chemotactic ability, and durability without affecting the anti-myeloma activity in vivo and in vivo, which provides strategies for overcoming the defects of low survival rate, poor durability and inhibition by tumor microenvironment of traditional CAR-T cells, and offers preliminary experimental basis for the clinical application of the fourth-generation CAR-T cells.

Objective:To design the fourth-generation chimeric antigen receptor-T (CAR-T) cells that secrete interleukin-7 (IL7) and chemokine C legend 19 (CCL19) on the basis of the second-generation CAR, and to analyze and compare the differences in proliferation, chemotaxis, tumor cell clearance and persistence in the microenvironment of multiple myeloma (MM) between them.Methods:The fourth-generation CAR vector plasmid was constructed by using 2A self-cleaving peptide technology. The third-generation lentiviral packaging system was used to prepare high-titer lentivirus. Flow cytometry was used to monitor the transduction efficiency of lentivirus and the subtype changes of CAR-T cells. The enzyme-linked immunosorbent assay (ELISA) was used to quantify the IL7 and CCL19 secreted by CAR-T cells.The calculation of absolute number of CAR-T cells during culture was used to analysis cell proliferation activity. Transwell migration assay was used to verify the chemotactic ability of CAR-T cells. The specific killing activity of CAR-T cells was detected by using the luciferase bioluminescence method. The NOD-Prkdcem26Cd52Il2rgem26Cd22/Nju (NOD) mouse xenograft model was used to verify the anti-myeloma activity and safety of CAR-T cells in vivo.Results:Flow cytometry results showed that the stable CAR expression rates of the second-generation anti-CS1 CAR-T and fourth-generation anti-CS1-IL7-CCL19 CAR-T cells were (91.50±0.29)% and (46.7±0.12)%, respectively. CAR-T cells were successfully constructed. Subtype analysis demonstrated that the ratio of stem memory T cell (TSCM) in anti-CS1-IL7-CCL19 CAR-T cells was (67.58±0.59)%, which was significantly higher than (50.74 ± 1.01)% of anti-CS1 CAR-T ( P=0.000 1), with more strong immune memory function and better durability. Anti-CS1-IL7-CCL19 CAR-T cells can continuously secrete IL7 and CCL19 compared to MOCK-T and anti-CS1 CAR-T ( P<0.000 1). The number of anti-CS1-IL7-CCL19 CAR-T cells reached (22.77±0.79)×10 6 on the 9th day after lentivirus transduction, which was significantly higher than (9.40±0.79)×10 6 of anti-CS1 CAR-T cells ( P=0.000 1), with stronger proliferation ability. The number of chemotaxis cells of anti-CS1-IL7-CCL19 CAR-T cells to reactive T cells was (109.0±4.04), which was significantly higher than (9.33±1.20) of MOCK-T ( P<0.000 1) and (7.33±0.88) of anti-CS1 CAR-T ( P<0.000 1), with stronger chemotactic ability. The specific killing activity showed that both anti-CS1-IL7-CCL19 CAR-T and anti-CS1 CAR-T cells had specific killing efficacies when compared with the MOCK-T cells ( P<0.000 1). Animal experiment indicated that anti-CS1-IL7-CCL19 CAR-T cells significantly reduced the tumor burden ( P<0.000 1) and extended the overall survival time ( P=0.006 1) of tumor-bearing mice. Conclusions:The anti-CS1-IL7-CCL19 CAR-T cells designed in this study show stronger proliferative activity, chemotactic ability, and durability without affecting the anti-myeloma activity in vivo and in vivo, which provides strategies for overcoming the defects of low survival rate, poor durability and inhibition by tumor microenvironment of traditional CAR-T cells, and offers preliminary experimental basis for the clinical application of the fourth-generation CAR-T cells.