Objective: To examine the effect of temperature on the risk of varicella in Lu'an City, Anhui Province, so as to provide insights into varicella prevention and control. Methods: Data on incidence of varicella in Lu'an City from 2010 to 2021 were collected from Chinese Disease Prevention and Control System, and meteorological data in Lu'an City were also collected from National Meteorological Science Data Center and China National Urban Air Quality Real-Time Publishing Platform during the same period. The effect of temperature on the risk of varicella was examined using a distributed lag non-linear model (DLNM) and subgroup analyses for gender and age were conducted. The effects of extremely low and high temperatures on the cumulative risk of varicella and trends in the cumulative risk of varicella over time were analyzed using a time-varying DLNM. Results: Totally 25 318 varicella cases were reported in Lu'an City from 2010 to 2021, including 15 013 men (59.30%) and 10 305 women (40.70%). The median number of varicella cases was 4 (interquartile range, 6) cases, and the daily median air temperature was 17.50 (interquartile range, 15.80) ℃, with the lowest temperature recorded as -5.80 ℃ and the highest temperatures as 34.90 ℃. The results from the DLNM showed that the extremely low temperatures reduced the risk of varicella (RR=0.522, 95%CI: 0.375-0.728) in relative to median temperature, while extremely high temperature increased the risk of varicella (RR=1.604, 95%CI: 1.112-2.316). Subgroup analysis revealed the effect curve for men was similar to total populations (extremely low temperature: RR=0.497, 95%CI: 0.331-0.746; extremely high temperature: RR=1.978, 95%CI: 1.260-3.106), and the effect of temperature on varicella risk was mainly concentrated among children at ages of 6 to 12 years (extremely low temperature: RR=0.426, 95%CI: 0.247-0.736; extremely high temperature: RR=2.431, 95%CI: 1.378-4.288). The results from the time-varying DLNM revealed that the cumulative risk of varicella due to both extremely low and high temperatures appeared a tendency towards a rise over years (P<0.05). Conclusions Low temperature may reduce the risk of varicella, while high temperature increases the risk of varicella in Lu'an City, which is more remarkable among men and children at ages of 6 to 12 years. The cumulative risk of varicella at both extremely low and high temperatures shows a tendency towards a rise over years.

Objective To determine the effect of Mycobacterium tuberculosis spp. inducing transformation of THP-1 cells to epithelioid cells (EC) and the involved signaling pathways and their regulations. Methods THP-1 cells infection models respectively infected with M. tuberculosis strains H37Rv, bovis and phlei were established. Indirect immunofluorescent staining assays were used to detect the expressions of monocyte/macrophage differentiation antigen CD115 and EC differentiation antigen CD82 of the THP-1 cells before or after infection. By Sandwich ELISA Kits, the phosphorylation levels of p38MAPK, Akt1 and STAT3 of the THP-1 cells before or after infection were measured. The alterations of CD115 and CD82 expression levels were examined when the associated signaling pathways were blocked with specific blocking agents. Results CD115 expression was weakened and CD82 expression was strongly increased in all the THP-1 cells infected with the three strains. A temporal up-regulation of the p38MAPK phoshporylation level but no obvious alteration of Akt and STAT3 phosphorylation levels after THP-1 cells infected by strain H37Rv or bovis. The THP-1 cells infected with anyone of the three strains continuously expressed CD115 after MAPK, PI3K/Akt or JAK/STAT of the cells was blocked. Although JAK/STAT was blocked, the THP-1 cells respectively infected with the three different strains still expressed CD82. However, CD82 expressed in THP-1 cells infected by the strain H37Rv or bovis was disappeared when p38MAPK and PI3K/Akt pathways of the cells were blocked. Conclusion Strain H37Rv and bovis can induce the infected THP-1 cells transforming to EC and p38MAPK and PI3K/Akt signaling pathways participate and regulate the transformation procedure. Of the two signaling pathways p38MAPK seems to be more important.

Objective:To study the correlation between hypercoagulant status of patients with hip fracture and the level of microparticles (MPs) in their peripheral blood, and to explore the feasibility of removing MPs to correct hypercoagulant status in patients with hip fracture.Methods:Sixty-five patients from December 2018 to September 2019 with hip fracture were included. There were 24 males and 41 females with the average age of 75.6±9.8 years old (range 58-95 years). Among them, 27patients (43.1%) were femoral neck fracture and38 patients (56.9%) were intertrochanteric fracture.All patients were diagnosed with X-ray and CT. Meanwhile, about 20 healthy people in the physical examination center included as controls in our study. There were 8 males and 12 females with the average age of 72.3±6.5 years old (range 56-81years). 2 ml of anticoagulant whole blood was taken on an empty body in the morning, and purified microparticlesby whole-blood density gradient centrifugation in whole blood were identified by electron microscope and nanoparticle tracking analyzer. After cell free plasma (CFP) was obtained by whole-blood density gradient centrifugation, the number of whole annexin V (AV) positived MPs and these MPs which from platelet (PMPs) was determined by flow cytometry. The activated clotting time (ACT) was determined by coagulation and platelet function analyzer to evaluate the degree of hypercoagulability. Then, Logistic analysis was performed on risk factors associated with hypercoagulability to determine whether the level of MPs was an independent risk factor for hypercoagulability, and the correlation between ACT value and MPs level was analyzed. Finally, the four coagulation items of each sample CFP before and after MPs removal were determined by automatic coagulation analyzer.Results:Under electron microscopy, MPs presented vesicular appearance,with a complete double-layer membrane structure, the size was in the range of 100-1 000 nm, and the morphology was not uniform. there were irregular vesicular and circular vesicular general shapes. The average size of MPs in peripheral blood of patients with hip fractures was 239.7±4.0 nm. The mean size of MPs distribution in the control group was 247.7±3.3 nm, and there was no statistically significant difference in MPs diameter between the two groups. The average level of circulating AV +MPs in patients with hip fracture was 564±171/μl, and the average level of PMPs was 326±104/μl. In the control group, the average level of AV +MPs was 252±82/μl, the average level of PMPs was 192±41/μl, the difference between AV +MPs and PMPs was statistically significant ( P<0.05). The average ACT level of patients with hip fracture was 324±94 s, while the average ACT level of the normal population was 535±76 s, and the difference between the two was statistically significant ( P<0.05). Single factor logistic regression analysis showed that the levels of APTT, PMPs and AV +MPs may be risk factors for hypercoagulability, and multivariate logistic regression analysis showed that AV +MPs is an independent risk factor for hypercoagulability.It has a highly negative correlation with ACT ( r=-0.822, P<0.05). The results of four coagulation items determined by CFP were PT 10.8±0.46 s, APTT 30.6±1.56 s, Fib 3.08±0.36 g/L, INR 0.98±0.04 and TT 19.3±0.62 s. After the removal of MPs, the coagulation function was PT 10.8±0.52 s, APTT 32.4±3.0 s, Fib 2.90±0.33 g/L, INR 0.99±0.05 s, and TT 19.9±0.63 s. There was no statistically significant difference before and after coagulation function. Conclusion:There is a hypercoagulable state in patients with hip fracture, moreover, the level of AV +MPs is an independent risk factor for hypercoagulability, which is highly correlated with ACT, and MPs has no significant effect on the classic four factors of coagulation.

Objective To investigate the incidence and related risk factors of delirium in elderly patients with hip frac-ture. Methods From October 2014 to February 2017, 306 patients with femoral neck fracture or intertrochanteric fracture more than 65 years old underwent elective hip arthroplasty (hip arthroplasty, proximal femoral nail fixation) were retrospective-ly analyzed. Patients' general information was recorded: age, sex, body mass index (BMI), history of diabetes mellitus, history of coronary heart disease, cerebral infarction, preoperative biochemical tests, left ventricular ejection fraction, thrombosis of both lower extremities, intraoperative status (ASA score, NYHA heart function score, anesthesia, anesthesia time, etc.), periop-erative blood transfusion, biochemical examination on the first day after surgery, perioperative albumin consumption, etc.; ac-cording to whether delirium happened after surgery, those patients were divided into delirium group and non-delirium Group and analyzed the incidence of postoperative delirium. Using univariate logistic regression analysis screened factors associated with postoperative delirium. The results were entered into the multivariate logistic regression analysis, screening the high risk factors for postoperative delirium.Results The incidence of delirium was 59(19.29%) after perioperative hip fracture, includ-ing 23 males and 36 females, ranging in age from 70 to 93 years with an average of (81.9±5.4) years. 247 cases had no deliri-um,including 81 males and 166 females,aged 65-96 years,with an average(76.4±8.1)years.The factors of age,length of hos-pital stay, days before surgery, preoperative hematocrit, perioperative albumin consumption, perioperative blood transfusion, preoperative leukocyte levels, preoperative albumin levels, preoperative creatinine, urea level, Mallampati classification (Mar-kov grade), cardiac function score (NYHA score), ASA score, intraoperative blood loss, postoperative leukocyte level, left ven-tricular ejection fraction, previous cerebral infarction, history of coronary heart disease, and diabetes were statistically signifi-cant different; univariate logistic regression analysis showed that the factors of age, ASA score, history of diabetes mellitus, length of hospitalization, and perioperative albumin consumption were demonstrated to have relationship with the occurrence of postoperative delirium. Multivariate regression analysis showed that age, history of diabetes and ASA score are the high risk factors of postoperative delirium.Conclusion Age,history of diabetes mellitus and ASA score are risk factors of delirium in el-derly patients with hip fracture.Those patients older than 75 years,with the history of diabetes mellitus or ASA score> 2 points were at higher risk of postoperative delirium,which prolongs hospitalization and increases protein consumption.

Objective To investigate reflectance confocal microscopy (RCM) features of irritant cutaneous reactions to sodium lauryl sulphate (SLS) in healthy persons aged from 18 to 60 years,to analyze effects of age and gender on cutaneous reactions,and to estimate the value of RCM in objective evaluation of cutaneous reactions.Methods An occlusive patch test was performed on the back of 120 healthy testees with 0.1％ and 0.5％ SLS solution (0.1％ and 0.5％ SLS groups) and distilled water (negative control group) for 48 hours.At different time points after the patch removal,clinical evaluation and RCM were performed.Results RCM imaging in the 0.1％ and 0.5％ SLS groups showed parakeratosis,indistinct structure of the stratum corneum,spongiosis and infiltration of inflammatory cells in the epidermis,and telangiectasia in the papillary dermis.The incidence of RCM features reached the peak until 24 hours after the removal of 0.1％ and 0.5％ SLS patches,and the incidence of telangiectasia in the dermis was up to 66.7％ and 95.0％ in the 0.1％ and 0.5％ SLS groups respectively.At 24 hours after the removal of 0.5％ SLS patch,the incidence of spongiosis was significantly lower in the males than in the females (68.9％ [42/61] vs.84.7％ [50/59],x2 =4.24,P ＜ 0.05).However,the incidence of spongiosis was significantly higher in testees aged 18-40 years than in those aged 41-60 years at 24 hours after the removal of 0.1％ SLS patch (53.3％[32/60] vs.35.0％[21/60],x2 =4.09,P ＜ 0.05).For the other RCM features,there were no significant differences in their incidence between different genders or age groups after the removal of 0.1％ and 0.5％ SLS patches (all P ＞ 0.05).Clinical evaluation showed that after the removal of 0.1％ and 0.5％ SLS patches,no significant difference in the incidence of irritant cutaneous reactions was observed between the males and the females or between the testees aged 18-40 years and those aged 41-60 years (all P ＞ 0.05).There were good correlations between the clinical evaluation results and RCM features.At 24 hours after the removal of 0.1％ SLS patch,the correlation coefficient between spongiosis and clinical evaluation results was up to 0.77,so was that between telangiectasia in the dermis and clinical evaluation results (both P ＜ 0.001).However,at 0.5 hour after the removal of SLS patches,clinical evaluation showed that the positive reaction rates were 2.5％ (3/120) and 12.5％ (15/120) in the 0.1％ and 0.5％ SLS groups respectively.In the meantime,there were 17.5 ％ (21 / 120) and 51.7％ (62/120) of testees manifesting more than 2 RCM features in the 0.1％ and 0.5％ SLS groups respectively,which were more similar to the clinical evaluation results at 24 hours after the removal of SLS patches (34.2％ [41/120] and 85.0％ [102/120] in the 0.1％ and 0.5％ SLS groups respectively) compared with the clinical evaluation results at 0.5 hour after the removal of SLS patches.Conclusions Neither gender nor age affects irritant cutaneous reactions to 0.1％ and 0.5％ SLS.Compared with clinical evaluation,RCM can evaluate cutaneous reactions more objectively and accurately in the early stage of irritant reactions.

Objective To investigate the role and mechanism of nico-tinamide adenine dinucleotide phosphate oxidase 4 (NAPHD oxidase 4,Nox4)-mediated reactive oxygen species (ROS) generation on high-dose dexamethasone (DEX) induced apoptosis in osteoblasts.Methods According to culture conditions,3rd passage of murine osteoblastic MC3T3-E 1 cells were divided into control group,Dexamethasone group,Dexamethasone+NAC (N-acetyl-L-cysteine) group,NAC group,Dexamethasone+DPI (Diphenyleneiodonium) group and DPI group.24 hours after culture,the morphology of osteoblasts was observed by inverted phase contrast microscope.Cell viability was determined by MTT assay.The generation of ROS in osteoblasts was measured using a fluorescent probe DCFH-DA.The apoptosis of each group was observed through Hoechst staining.The mRNA level and protein expression of Nox4 were detected by real-time quantitative PCR and Western Blot.In addition,after silence of Nox4 with small interfering RNA (siRNA),the ROS generation was further detected by a fluorescent probe DCFH-DA.Results After treatment with 1000 nmol DEX for 24 hours,compared to control group,the results of inverted phase contrast microscope and MTT showed that osteoblasts in DEX group exhibited more obvious signs of shrinkage and deformation with decreased cell viability.After intervene with NAC and DPI,morphology of osteoblasts was good with increased viability of osteoblasts.Compared to control group (5.86％± 0.28％),the production of ROS in DEX group (45.14％±1.49％) was significantly increased (P=0.000).The apoptotic rates in DEX group (29.60％± 1.52％) was significantly increased compared with control group (4.12％±0.67％) (P=0.000).Compared to DEX group,the production of ROS generation in DEX+NAC group (28.06％±1.61％) and DEX+DPI group (23.70％±1.28％) was significantly decreased (P=0.000).It presented that NAC or DPI significantly decreased the formation of ROS.Compared to DEX group,the apoptotic rate in DEX+NAC group (8.94％± 1.47％) and DEX+DPI group (12.96％±2.03％) was significantly decreased (P=0.000).It presented that NAC or DPI significantly decreased osteoblast apoptosis.In addition,the Nox4 mRNA level in DEX group was 2.67-fold compared with control group (t=-10.301,P=0.009).The difference had statistically significance.The protein expression of Nox4 in DEX group was 2.37-fold compared with control group (t=-15.542,P=0.004).The difference has statistically significance.After silence of Nox4 by siRNA,the generation of ROS in DEX+Nox4 siRNA group (14.53％± 1.00％) was decreased by 16.92％ compared with DEX group 31.45％±0.72％ (P=0.000).The difference had statistically significance.Conclusion Nox4-mediated ROS generation plays an important role in osteoblasts apoptosis induced by high-dose dexamethasone.It provided us the new target in the management of Nox4 to provide possible therapy for steroid-induced avascular necrosis of the femoral head (SANFH).

Objective To test the antimicrobial susceptibility of Staphylococcus aureus from children with impetigo,and to assess the differences in randomly amplified polymorphic DNA profiles between sensitive and resistant Staphylococcus aureus strains.Methods Secretion specimens were obtained from the impetiginous lesions of 178 children,and subjected to bacterial culture.The susceptibility of 162 Staphylococcus aureus isolates against 21 antibiotics was tested.Randomly amplified polymorphic DNA PCR(RAPD-PCR)was performed to characterize the genotype of Staphylococcus aureus.Results Totally,180 bacterial strains were isolated from 178 children with impetigo in Chengdu,including 162(90.00％)Staphylococcus aureus strains.Of the 162 Staphylococcus aureus strains,148 were methicillin sensitive Staphylococcus aureus(MSSA),14 methicillin resistant Staphylococcus aureus(MRSA).The most active antibiotic was minocycline,followed by teicoplanin,quinupristin,vancomycin and nitrofurantoin,while the resistance rate to penicillin was highest,followed by that to erythromycin,clindamycin,compound sulfamethoxazole and tetracycline.All the Staphylococcus aureus isolates were sensitive to fusidic acid,nitrofurantoin,vancomycin,minocycline and teicoplanin.According to RAPD-PCR,the 162 Staphylococcus aureus strains were divided into 8 genotypes,with the three most prevalent genotypes being Ⅲ(31.48％),Ⅱ(26.54％)and Ⅵ(25.93％),which accounted for 65.43％(106/162)in all the strains.The 148 MSSA strains fell into 8 genotypes,with genotype Ⅲ(50 strains,33.78％),Ⅵ(39 strains,26.35％)and Ⅱ(33 strains,22.30％)being the most prevalent genotypes;the 14 MRSA strains fell into 3 genotypes,i.e.,genotype Ⅱ(10 strains,71.43％),Ⅵ(3 strains,21.43％),and Ⅲ(1 strain,7.14％).Conclusions Staphylococcus aureus is the most prevalent pathogenic bacteria in children with impetigo in Chengdu area,which is highly sensitive to minocycline,teicoplanin and quinupristin,and falls into 8 genotypes according to RAPD-PCR with genotype Ⅲ being the most common genotype.

Objective:To establish a hypergravity loading model with a high-acceleration centrifugal loading device and to investigate the effects of different hypergravity loading and icariin on osteoblast adhesion and cytoskeleton.Methods:MC3T3-E1 cells were seeded in the dishes of cell culture at a density of 2×10 5/cm 2. And the experiment was divided into 6 groups: control group (without icariin and loading); simple administration group (only icariin); 10 G loading group (only loading); 10 G administration group (with icariin and loading); 40 G loading group (only loading); 40 G administration group (with icariin and loading). The experimental loading group was loaded with MC3T3-E1 cells using a high-acceleration centrifugal loader. And continuous loading for 3 d, 30 min per d. The control group and the simple administration group were exposed to normal gravity, and the remaining conditions were not different from the experimental group. Icariin was used at a concentration of 10 -7 mol/L in all administration groups, and the experiments were carried out according to the method of preventive administration. At the same time, the related molecular biological techniques such as alizarin red staining, alkaline phosphatase (ALP) activity measurement, CCK-8 cell proliferation experiment, cytoskeleton phalloidin staining, qPCR and Western Blot were used to detect the effects of icariin on osteoblasts adhesion protein integrin α5 and integrin β1 and cytoskeleton protein F-actin under hypergravity extreme mechanical environment. Results:All models were successfully prepared. The alizarin red staining: The icariin could significantly promote the formation of osteoblastic calcified nodules. And the 10 G loading could also promote the mineralization of osteoblasts and increase the number of mineralized nodules, while the mineralization and the number of mineralized nodules of osteoblasts are significantly reduced in 40 G loading. ALP activity test: The OD values of simple administration group, 10 G loading group and 40 G loading group were 0.246, 0.331 and 0.163, respectively. Compared with 0.207 in the control group, the differences were statistically significant ( P<0.05). The 10 G administration group and the 40 G administration group were 0.373 and 0.180, and the differences were statistically significant ( P<0.05). The results of CCK-8 proliferation experiments: The OD value of simple administration group were 0.650, which was statistically significant compared with 0.551 of control group ( P=0.031). The 10 G loading group and 40 G loading group were 1.193 and 0.245, and their differences with the control group were both statistically significant ( P<0.05). The OD value of 10G administration group and the 40 G administration group were 1.300 and 0.310, which were significantly different from the respective loading groups ( P<0.05). Phalloidin staining: 10 G loading significantly increased the number of cells, but the changes in cells morphology and skeleton were not obvious. 40 G loading significantly inhibited the increase of the number of cells, meanwhile, made the pseudopods of cells more shorter and even disappeared. 40 G loading made the seriously damage of the cytoskeleton and even cause the cells to death. Icariin had no effect on the cells morphology, but it did has a certain repair effect after the cells loading. The results of qPCR and Western Blot experiments all confirmed that the expressions of integrin α5, integrin β1 and F-actin were up-regulated after icariin treatment. 10 G loading could promote the expression of integrin α5, integrin β1 and F-actin, and 40 G loading significantly inhibited the expression of the mRNA and proteins. Conclusion:Both 10 G condition and icariin can promote the development, cell adhesion and the cytoskeleton's stability of osteoblasts, while 40 G has a significant inhibitory effect.

OBJECTIVE To investigate the mechanism of glucose -6-phosphate dehydrogenase （G6PD）-induced sorafenib - resistance in hepatocarcinoma cell based on phoshorylated 3-kinase/protein kinase B （PI3K/Akt）signaling pathway . METHODS Cell lines including hepatocarcinoma cell Huh 7，sorafenib-resistant cell Huh 7-SR，G6PD overexpressed cell Huh 7-G6PD and its control cell Huh 7-CT，and compounds including G 6PD inhibitor （6-Aminonicotinamide，6AN）and sorafenib were used as objects or intervention drugs in these research . CCK8 assay was applied to evaluate cell viability . The protein levels of G 6PD and the phosphorylation levels of PI 3K/Akt signaling pathway were detected by Western blot . Flow cytometry was utilized to investigate cell apoptosis. RESULTS Compared with Huh 7 cells，the protein level of G 6PD was significantly increased in Huh 7-SR cells （P＜ 0.05）. The combination of 6AN and sorafenib reduced cell viability of Huh 7-SR cells （P＜0.01）. However，compared with Huh 7- CT，increased cell viability and decreased cell apoptosis rate were observed in Huh 7-G6PD cells while cells were treated with sorafenib（P＜0.01）. Mechanistically，the phosphorylation levels of PI 3K and Akt were significantly decreased in Huh 7-SR cells that were treated with 6AN（P＜0.05）. Moreover，under the condition of no drug intervention ，the phosphorylation levels of PI 3K and Akt were significantly elevated in Huh 7-G6PD cells when compared with Huh 7-CT（P＜0.01）. CONCLUSION G6PD could induce sorafenib -resistance in hepatocarcinoma cell by activating PI 3K/Akt signaling pathway .

OBJECTIVE To investigate the regulatory effect of autophagy on the resistance of human liver cancer cell Huh7 to lenvatinib. METHODS Using human liver cancer cell Huh7 as subject， the lenvatinib-resist cell model （Huh7-LR） was generated by the low-dose gradient method combined with long-term administration. The sensitivity of parental cell Huh7 and drug-resistant cell Huh7-LR to lenvatinib was detected by using CCK-8 assay and flow cytometry. Western blot assay and GFP-mCherry-LC3 plasmid transfection were performed to detect the expression levels of autophagic protein Beclin-1， autophagic adapter protein sequestosome 1 （p62）， microtubule-associated protein 1 light chain 3 （LC3） and autophagic level. Furthermore， an autophagy activation model was constructed by cell starvation， the protein expression of p62 and autophagy level were detected by using Western blot assay and GFP-mCherry-LC3 plasmid transfection， and the effect of autophagy activation on the sensitivity of Huh7-LR cells to lenvatinib was detected by flow cytometry. RESULTS Compared with parental cells， the drug resistance index of Huh7-LR cells was 6.2； protein expression of p62 was increased significantly， while apoptotic rate， protein expression of Beclin-1 and LC3Ⅱ/ LC3Ⅰ ratio were all reduced significantly （P＜0.05 or P＜0.01）； the level of autophagy was decreased to some extent. Autophagy activation could significantly increase the protein expression of p62 in Huh7-LR cells （P＜0.05） and autophagy level， and significantly increase its apoptotic rate （P＜0.05）. CONCLUSIONS Autophagy is involved in lenvatinib resistance， and activating autophagy can reverse the resistance of liver cancer cells to lenvatinib to some extent.