Objective To study the effects of semaphorin 5A (SEMA5A) gene silencing by lentivirus?mediated short hairpin RNA(shRNA)on biological activity of malignant melanoma cell line A375. Methods Two pairs of interference sequences for SEMA5A gene(shRNA1 and A375?shRNA2)and a pair of control interference sequences were designed to build lentiviral vectors, which were then transfected into HEK293T cells to gain lentivirus. A375 cells were divided into three groups:experimental group(A375?shRNA1 and A375?shRNA2 cells)transfected with the lentivirus containing shRNA1 or shRNA2, negative control group (A375?con cells) transfected with that containing the control shRNA, and blank control group(A375 cells)receiving no transfection. The A375 cells with stable knockdown of SEMA5A gene expression were screened by puromycin. Subsequently, reverse transcription?PCR and Western?blot analysis were performed to detect mRNA and protein expressions of Semaphorin 5A in these cells, and methyl thiazolyl tetrazolium(MTT)assay was applied to evaluate the growth of cells. The scratch assay and invasion assay were conducted to estimate migration and invasion ability of cells. Results The lentivirus containing the SEMA5A?targeting shRNAs or control shRNA was successfully transfected into A375 cells, and stably transfected cells were gained after puromycin selection. The expressions of semaphorin 5A mRNA and protein in the A375?shRNA2 cells were significantly reduced compared with those in the A375?con and A375 cells(all P < 0.05). MTT assay showed that the growth of A375?shRNA2 cells was significantly slower than that of A375?con and A375 cells(both P<0.05), while there was no significant difference in the growth rate between A375?con and A375 cells(P>0.05). The scratch assay showed that there was no obvious cell migration into the scratch in the experiment group, whereas the scratch was almost covered by cells in the negative control group and blank control group. The invasion assay showed that the number of A375?shRNA2 cells passing through the Transwell chamber was significantly smaller than that of A375 and A375?con cells(both P<0.05), while there was no significant difference between that of A375 and A375?con cells(P > 0.05). Conclusion The silencing of SEMA5A gene by lentivirus?mediated shRNA could effectively down?regulate the expression of semaphorin 5A, and inhibit the growth, invasion and migration of A375 cells.

Objective: To explore the acute toxicity of Diquat in mice and to calculate the median lethal dose (LD₅₀) of Diquat to rats and observe the pathological changes of tissues and organs in rats with different concentrations of Diquat. Methods: Diquat solution of 50 mg/kg was prepared freshly with 1 000 mg of Diquat and dilute the solution with water to a total of 20 ml. A total of 99 healthy adult male Wistar rats were randomly divided into part one, part two and control groups. In the first part, 36 rats were randomly divided into 4 groups: 100 mg/kg group, 200 mg/kg group, 300 mg/kg group and 400 mg/kg group, which were treated with 100 mg/kg, 200 mg/kg, 300 mg/kg and 400 mg/kg of Diquat solution by gavage, respectively. The death and symptoms of poisoning after intragastric administration were recorded, and the maximum tolerated dose and absolute lethal dose were measured. In the second part, 54 rats were randomly divided into 6 groups: 200 mg/kg group, 220 mg/kg group, 240 mg/kg group, 260 mg/kg、280 mg/kg group and 300 mg/kg group, whichwere treated with 200 mg/kg, 220 mg/kg, 240 mg/kg, 260 mg/kg, 280 mg/kg and 300 mg/kg of Diquat solution by gavage, respectively. The survival of rats in different concentration of Diquat was observed and the LD₅₀ was calculated by Excel processing the formula of Koch's method. The control group were given equal volume water under the same experimental conditions. And moreover, the lungs, kidneys, hearts, livers, and brain tissues were collected and fixed by formaldehyde, embedded by paraffin, and sectioned for histopathological light microscopy. Results: The maximum tolerated dose was 240 mg/kg and the absolute lethal dose was 300 mg/kg. The LD₅₀ of Diquat for Rats was 280.58 mg/kg. The high-dose group had significantly more organ damage than the low-dose group after diquat poisoning. Conclusion The determination of the half-lethal dose of diquat, at the same time observed multiple organs damaged in rats after the diquat quickly poisoned. Kidneys, lungs and heart might be the main organ which was heavily damaged. With the extension of observation time, the organ damage of rats exposed to small doses gradually stabilized.

Objective:To correlate acute ischemic stroke with leukoaraiosis with intracranial and extracranial artery stenosis.Methods:A total of 300 patients with acute ischemic stroke admitted to Shaoxing Second Hospital from January to December 2017 were included in this study. All patients underwent magnetic resonance (MRI) examination. According to the examination results, these patients were divided into control (acute ischemic stroke, n = 100) and acute ischemic stroke with leukoaraiosis, n = 200). Carotid artery plaque size and blood sugar level were recorded in each group. Intracranial and extracranial large artery stenosis rates were compared between the two groups. Severity of leukoaraiosis was correlated with intracranial and extracranial artery stenosis. Results:The percentage of patients developing hypertension in the observation group was significantly higher than that in the control group [66.0% (132/200) vs. 44.0% (44/100), χ2 = 13.31, P < 0.01]. The incidence of coronary heart disease in the observation group was significantly higher than that in the control group [49.0% (98/200) vs. 31.0% (31/100), χ2 = 8.81, P < 0.01]. The incidence of carotid artery plaque in the observation group was significantly higher than that in the control group [49.5% (99/200) vs. 34.0% (34/100), χ2 = 6.49, P = 0.01]. The incidence of carotid artery stenosis in the observation group was significantly higher than that in the control group [23.5% (47/200) vs. 12.0% (12/100), χ2 = 5.58, P = 0.01]. There was no significant difference in the incidence of anterior cerebral artery stenosis between observation and control groups [5.5% (11/200) vs. 4.0% (4/100), χ2 = 0.32, P = 0.57]. The size of carotid artery plaque in the observation group was significantly larger than that in the control group [(1.86 ± 0.42) cm vs. (1.39 ± 0.27) cm, t = 10.18, P < 0.01]. The incidence of intracranial and extracranial artery stenosis in the observation group was significantly higher than that in the control group [41.0% (82/200) vs. 24.0% (24/100), χ2 = 8.43, P < 0.01]. The severity of leukoaraiosis was positively correlated with the degree of intracranial and extracranial artery stenosis ( r = 0.79, P < 0.01). Conclusion:Patients with acute ischemic stroke with leukoaraiosis have a high intracranial and extracranial artery stenosis and the severity of leukoaraiosis is positively correlated with intracranial and extracranial artery stenosis.

Objective To determine the expression of caspase-14 in skin lesions of patients with chronic actinic dermatitis (CAD),and to explore the effect of ultraviolet B (UVB) radiation on its mRNA and protein expression in HaCaT cells.Methods In 2016,skin samples were collected from lesions of 10 patients with CAD (test group),10 patients with eczema (positive control group) and from normal skin of 10 healthy controls after cosmetic surgery (negative control group) in the Department of Dermatology,First Affiliated Hospital of Kunming Medical University.Immunohistochemical staining was performed to determine the expression of caspase-14 in the normal skin,CAD and eczema lesions.Cultured HaCaT cells were divided into several groups:UVB groups irradiated with 0,30,60,90 mJ/cm2 UVB separately,and 5-AzaC groups irradiated with 0,30,60,90 mJ/cm2 UVB separately followed by the treatment with the methylase inhibitor 5-AzaC for 24 hours.Then,the cells were collected,and real-time fluorescence-based quantitative PCR (RT-PCR) and Western blot analysis were conducted to determine the mRNA and protein expression of caspase-14 respectively in HaCaT cells in the UVB groups and 5-AzaC groups.Statistical analysis was carried out with SPSS22.0 software by using chi-square test for the comparison of rates,and t test and two-factor analysis of variance for the comparison of means.Results In the CAD and eczema lesions,caspase-14 was mainly expressed in the spinous and granular layers,but not in the stratum comeum.However,caspase-14 was markedly expressed in the stratum corneum of the normal skin tissues.Of the 10 CAD samples,5 were positive for caspase-14,and 9 of 10 normal skin samples were positive for caspase-14.The positive rate of caspase-14 significantly differed between the two above groups (x2 =7.30,P ＜ 0.05).RT-PCR and Western blot analysis showed significant changes in the mRNA and protein expression of caspase-14 in HaCaT cells after irradiation with different doses of UVB (F =87.54,23.46,both P ＜ 0.05),which showed a decreasing trend along with the increase in the dose of UVB.After exposure to 0,30,60 and 90 mJ/cm2 UVB,the mRNA and protein expression of caspase-14 was significantly higher in the 5-AzaC groups than in the UVB groups (all P ＜ 0.05).Conclusions In CAD lesions,the expression of caspase-14 markedly decreased,and was absent in the stratum corneum.UVB radiation can downregulate the mRNA and protein expression of caspase-14 in HaCaT cells.

Objective:To investigate various clinical treatment measures for thoracolumbar fractures and provide new diagnosis and treatment methods.Methods:The case data of 5 patients(3 males, 2 females, and the age ranged from 39 to 59 years with an average of 51 years) with thoracolumbar fractures were treated with Waveflex semi-rigid internaI fixation system from May 2020 to December 2021 were retrospectively analyzed, imaging indexes and clinical effects were followed up, and analyzed and summarized in combination with relevant literatures.Results:The operations of all 5 patients were successfully completed, and the follow-up period was 6 months. At the last follow-up, the internal fixation position was good, the height of the injured vertebra was maintained satisfactorily, and the clinical effect was satisfactory.Conclusion:Waveflex semi-rigid internal fixation system combines the advantages of fusion and non-fusion, providing a new idea for the treatment of thoracolumbar fractures, but its long-term effect still needs further follow-up.

Objective We planned to evaluate the effectiveness of moving epidemic method (MEM) in calculating influenza epidemic threshold of 7 climatic zones in China mainland. Methods The positive rate of influenza virus was obtained from the National Influenza Surveillance Network System from 2010/2011 to 2017/2018. We divided the 31 provinces into 7 climatic zones according to previous literatures and applied MEM to calculate the influenza epidemic threshold of 2018/2019 influenza season for these climatic zones. Sensitivity, specificity, positive predictive value and negative predictive value were calculated to evaluate the effectiveness of MEM. Results Pre?epidemic threshold (the positive rate of influenza virus) varied from 9.66% (temperate zone) to 16.36% (subtropical zone) for 2018/2019 influenza season. The gap between pre?epidemic and post?epidemic thresholds was less than 5% except for plateau zone. The sensitivity was 86.16% (95CI: 66.81% - 98.23%), the specificity was 94.92% (95CI:91.13%-98.41%), the positive predictive value was 89.87% (95% CI : 84.39%-94.38%), the negative predictive value was 92.96% (95%CI: 84.46%-99.17%). Conclusion Overall, moving epidemic Method performs well in calculating influenza epidemic threshold in China, much better than the previous study.

Objective We planned to evaluate the effectiveness of moving epidemic method (MEM) in calculating influenza epidemic threshold of 7 climatic zones in China mainland. Methods The positive rate of influenza virus was obtained from the National Influenza Surveillance Network System from 2010/2011 to 2017/2018. We divided the 31 provinces into 7 climatic zones according to previous literatures and applied MEM to calculate the influenza epidemic threshold of 2018/2019 influenza season for these climatic zones. Sensitivity, specificity, positive predictive value and negative predictive value were calculated to evaluate the effectiveness of MEM. Results Pre?epidemic threshold (the positive rate of influenza virus) varied from 9.66% (temperate zone) to 16.36% (subtropical zone) for 2018/2019 influenza season. The gap between pre?epidemic and post?epidemic thresholds was less than 5% except for plateau zone. The sensitivity was 86.16% (95CI: 66.81% - 98.23%), the specificity was 94.92% (95CI:91.13%-98.41%), the positive predictive value was 89.87% (95% CI : 84.39%-94.38%), the negative predictive value was 92.96% (95%CI: 84.46%-99.17%). Conclusion Overall, moving epidemic Method performs well in calculating influenza epidemic threshold in China, much better than the previous study.

OBJECTIVEProviding evidences for further modification of China Infectious Diseases Automated-alert and Response System (CIDARS) via analyzing the outbreak detection performance of Moving Percentile Method (MPM) by optimizing thresholds in different provinces.

METHODSWe collected the amount of MPM signals, response results of signals in CIDARS, cases data in nationwide Notifiable Infectious Diseases Reporting Information System, and outbreaks data in Public Health Emergency Reporting System of 16 infectious diseases in 31 provinces in Chinese mainland from January 2011 to October 2013. The threshold with the optimal sensitivity, the shortest time to detect outbreak and the least number of signals was considered as the best threshold of each disease in Chinese mainland and in each province.

RESULTSAmong all the 16 diseases, the optimal thresholds of 10 diseases, including dysentery, dengue, hepatitis A, typhoid and paratyphoid, meningococcal meningitis, Japanese encephalitis, scarlet fever, leptospirosis, hepatitis, typhus in country level were the 90(th) percentile (P90), which was the same as provincial level for those diseases.For the other 6 diseases, including other infectious diarrhea, influenza, acute hemorrhagic conjunctivitis, mumps, rubella and epidemic hemorrhagic fever, the nationwide optimal thresholds were the 80th percentile (P80), which was different from that by provinces for each disease. For these 6 diseases, the number of signals generated by MPM with the optimal threshold for each province was decreased by 23.71% (45 557), 15.59% (6 124), 14.07% (1 870), 9.44% (13 881), 8.65% (1 294) and 6.03% (313) respectively, comparing to the national optimal threshold, while the sensitivity and time to detection of CIDARS were still the same.

CONCLUSIONOptimizing the threshold by different diseases and provinces for MPM in CIDARS could reduce the number of signals while maintaining the same sensitivity and time to detection.

OBJECTIVETo analyze the implement performance of China Infectious Diseases Automated-alert and Response System (CIDARS) of 31 provinces in mainland China, and to provide the evidences for further promoting the application and improvement of this system.

METHODSThe amount of signals, response situation and verification outcome of signals related to 32 infectious diseases of 31 provinces in mainland China in CIDARS were investigated from 2011 to 2013, the changes by year on the proportion of responded signals and timeliness of signal response were descriptively analyzed.

RESULTSA total of 960 831 signals were generated nationwide on 32 kinds of infectious diseases in the system, with 98.87% signals (949 936) being responded, and the median (the 25(th) percentile to the 75(th) percentile (P25-P75) ) of time to response was 1.0 (0.4-3.3) h. Among all the signals, 242 355 signals were generated by the fixed-value detection method, the proportion of responded signals was 96.37% (62 349/64 703), 98.75% (68 413/69 282) and 99.37% (107 690/108 370), respectively, and the median (P25-P75) of time to response was 1.3 (0.3-9.7), 0.8(0.2-4.9) and 0.7 (0.2-4.2) h, respectively. After the preliminary data verification, field investigation and laboratory test by local public health staffs, 100 232 cases (41.36%) were finally confirmed.In addition, 718 476 signals were generated by the temporal aberration detection methods, and the average amount of signal per county per week throughout the country were 1.53, and 8 155 signals (1.14%) were verified as suspected outbreaks. During these 3 years, the proportion of signal response was 98.89% (231 149/233 746), 98.90% (254 182/257 015) and 99.31% (226 153/227 715), respectively, and the median (P25-P75) of time to response was 1.1 (0.5-3.3), 1.0 (0.5-2.9) and 1.0 (0.5-2.6) h, respectively.

CONCLUSIONFrom 2011 to 2013, the proportion of responded signals and response timeliness of CIDARS maintained a rather high level, and further presented an increasing trend year by year. But the proportion of signals related to suspected outbreaks should be improved.

Objective To learn the characteristics of morbidity and mortality of notifiable diseases reported in China in 2013.Methods Descriptive analysis method was used to analyze the morbidity and mortality of notifiable diseases in China in 2013,with Microsoft Excel 2010 and ArcGIS 10.0 used to develop statistical charts.Results In 2013,the morbidity of the nationwide notifiable diseases was 473.87/100 000,a decrease of 3％ below the average of the recent 3 years,while the mortality was 1.23/100 000,an increase of 2％ over the average of the recent 3 years.The rate of laboratory diagnosis of the reported cases was 38.4％.Top 5 diseases of the reported incidence were hand foot and mouth disease,other infectious diarrhea,hepatitis B,tuberculosis and syphilis.The death cases reported were mainly AIDS,tuberculosis and rabies.As classified by the transmission route analysis,intestinal infectious diseases accounted for 49％ of the total incidence reported for the year,followed by the blood and sexually transmitted infectious diseases,respiratory infectious diseases,animal and vector borne infectious diseases.According to the pathogenic analysis,virus infectious diseases accounted for 68％ of the total cases,higher than bacterial infectious diseases and animal-borne/vector-borne infectious diseases.In Shanghai,Zhejiang and Jiangsu,human infection with avian influenza A (H7N9) virus was emerging,as epidemic situation of measles,dengue fever and brucellosis were on the rise significantly nationwide,while the morbidity of infectious diseases decreased,namely pulmonary tuberculosis,hepatitis B among others.Morbidity of the top 5 provinces for notifiable infectious diseases were Hainan,Guangxi,Guangdong,Xinjiang and Zhejiang,respectively.Conclusion The proportion of laboratory confirmed cases among totals was still low in 2013.The morbidity of the infectious diseases was higher in western provinces and parts of south-eastern province,the mortality was higher in westem provinces.The emerging human infection with avian influenza A (H7N9) virus,and the high epidemic of measles,dengue fever and brucellosis in some areas had caught the society concerns.