Objective To interpret genetic variation and population structure of Anopheles dirus A and D from China by molecular marker. Methods Samples included An. dirus A of Hainan laboratory colony (n=13), and field specimen from Mengla (n=17) and Jiangcheng (n=17) in Yunnan Province. The specimens were identified by PCR assay before study. mtDNA-COⅠ region was amplified and sequenced. Genetic variation and population structure was estimated according to sequence data. Results The mtDNA-COⅠ gene with a length of 959 bp was analyzed. There were three haplotypes in An. dirus A and six haplotypes in An. dirus D. The above haplotypes distributed in three populations unif-ormly. The average number of pairwise differences within Mengla population (7.441 2) was greater than that of Jiangcheng (1.279 4) and Hainan (1.051 3) populations, which suggested that the level of genetic divergence was the highest within Mengla population. The result of hierarchical AMOVA estimation showed a limited geneflow (Fst=0.799 9), therefore the variation level in a population (20.01%) was smaller than among the populations (79.99%). Conclusion The inter-specific genetic variation between An. dirus A and D in China was small and the level of divergence among individuals was high.

Objective To establish the molecular identification of five members in Anopheles maculatus complex from China. Methods Different rDNA-ITS2 regions of An. maculatus complex were sequenced and analyzed. The species specific primers were designed, and PCR assay was used for the identification. Results The length and GC contents of ITS2 were 328 bp, 58.54% in An. pseudowillmori, 330 bp, 57.85% in An. maculatus, 337 bp, 59.05% in An. willmori, 334 bp, 58.68% in An. dravidicus, and 338 bp, 57.69% in An. sawadwongporni, respectively. The intra-species ITS2 sequences were conservative. The ranges of divergence level among five members were from 9.7% to 18.9% . Five distinct specific fragments were amplified by PCR assay using five species specific primers and 5. 8S primer. The length was 119, 186, 231, 327 and 406 bp respectively. Conclusion The diagnostic PCR assay based on ITS2 divergence to distinguish five members of An. maculatus complex was simple and reliable.

Objective To improve the differential diagnosis between autoimmune pancreatitis (AIP) and pancreatic cancer (PC) by a contrast analysis of imageological features.Methods Thirty-six patients who had postoperative pathological with Asian AIP standards and 95 patients who had postoperative pathological consistent with PC.The imageological results of these AIP and PC patients were analyzed.Results AIP was significantly less than PC in the enhanced CT of a mass or enlargement of the pancreatic head,enlargement of the lymph nodes around the pancreas,dilation and interrupt in pancreatic and bile duct,peripheral vascular and organ involvement (11/27 vs.28/40,2/27 vs.17/40,13/27 vs.32/40,1/27 vs.10/40,8/27 vs.26/40,2/27 vs.15/40,0/27 vs.15/40,0/27 vs.10/40,P ＜ 0.05).AIP was significantly more than PC in the enhanced CT of a diffusely enlarged pancreas,calcification or pancreatic calculus,capsule-like rim or the vague peripancreatic fat interval (4/27 vs.0/40,7/27 vs.0/40,10/27 vs.6/40,P ＜ 0.05).AIP was significantly less than PC in the three-dimensional ultrasonography of dilation diameter of pancreatic duct and dilation of common bile duct [(0.421 ± 0.270) cm vs.(0.594 ± 0.270) cm,1/18 vs.16/26,P＜ 0.05].AIP was significantly less than PC in the magnetic resonance cholangiopancreatography of dilation of common bile duct and interrupt in pancreatic duct (7/13 vs.16/18,1/13 vs.10/18,P ＜ 0.05).Conclusion AIP as a unique type of chronic pancreatitis can be distinguished from PC on distinctive imageological features

Objective To investigate the effects of early estrogen replacement therapy(ERT) with different doses on aortic endothelial senescence and its possible mechanism. Methods Twenty- eight healthy New Zealand white female rabbits were randomized into four groups: group A(n=7), in sham operation;group B(n=7), ovariectomized;group C(n=7), ovariectomized and in low-dose ERT(estradiol benzoate 200 μg,im, QOD) ;and group D(n=7), ovariectomized and in high-dose ERT (estradiol benzoate 1000 μg,im,QOD). All rabbits were fed with high fat diet and the rabbits in group C and group D were given ERT from the 7th day after the operation. Blood samples taken via ear central artery before the operation and after the high fat diet for 4 and 12 weeks were used to determine the concentrations of serum estradiol(E2), total cholesterol(TC), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), trigtyceride (TG) and asymmetric dimethylarginine(ADMA). After 12 weeks, the aorta was separated for histopathologic analysis and the areas of senescent endothelium and atherosclerotic plaque were calculated.Results (1)After ovariectomized, the concentration of serum estradiol(E2) was significantly lower in group B than in other groups (all P<0.05). Twelve weeks later, the level of E2in was higher group D than in group A and C(both P<0. 05), but there was no significant difference between group C and group A (P< 0. 05). (2)After the high-fat diet for 4 and 12 weeks, there was a notable increase of TC, LDL-C, TG and HDL-C levels in rabbits of each group(all P<0. 01). After 12 weeks, the levels of TC and LDL- C were remarkably higher in group B than in other groups(all P<0. 01), however, the levels of TG and HDL-C were lower in group B than in other groups(all P<0. 01). The concentrations of TC and LDL-C in group C and group D were lower than those in group A (all P<0.01), but the differences of the levels of TG and HDL-C in group C and group D were not significant in comparison with group A (all P>0. 05). There were no significant differences in the lipid levels between group C and D (all P> 0. 05). (3)After 12-week high fat diet, there were a striking increases of serum ADMA level in all four groups (all P>0. 05). The level of ADMA was higher in group B than in other groups (P< 0. 05) andlower in group C and D than that in group A (both P<0. 05). There was no significant difference between group C and D (P<0. 05). (4)12 weeks later, the areas of senescent endothelium and atherosclerotic plaque in group B were significantly larger than those in other groups (all P< 0. 01), and the areas were smaller in group C and D than those in group A (all P<0. 01), there was no significant difference between group C and group D(both P>0.05). (5)Linear correlation analysis showed that there was an obvious positive correlation between the areas of senescent endothelium and atherosclerotic plaque(r=0. 962, P<0. 01), and both of them were positively correlated with the levels of serum AMDA (r=0. 812,0. 824,0. 755,0. 797,0. 749,0. 727), TC (r=0. 812,0. 824) and LDL-C(r=0. 755,0. 797) (all P<0. 01) and negatively correlated with the concentration of serum E2 (r=-0. 762,-0. 743, both P<0. 01).Conclusions Early ERT can improve serum lipid metabolism, reduce ADMA level, inhibit vascular endothelial senescence and attenuate atherogenesis. The delaying effect of estrogen on endothelial senescence may be due to its improving lipid metabolism and lowering ADMA level.

Objective To establish the enzyme-linked immunosorbent assay (ELISA) methods for the quantitative determination of IgG antibodies against diphtheria (DT) and tetanus (TT).MethodsPurified diphtheria toxiod and tetanus toxoid were respectively used as the coating antigens,the human-derived serum antibody standard substance of DT and TT served as the standard substance.The dose-response curves of the tested samples and standard substance were fitted.Then the two quantitative ELISA methods for determining the antibody to DT (Anti-DT) and antibody to TT (Anti-TT) were established with the parallel lines method.Then the methodological verification and application study were conducted.Results The validation results of the two quantitative ELISA measurement methods were in accordance with the regulations.The quantity limit of ELISA method for quantitative detection of Anti-DT demonstrated to be 0.084 mIU/mL,its average recovery rate was 97.6%.The intra-assay coefficient of variation(CV) and inter-assay CV of this Anti-DT assay were ≤ 3.40% and ≤5.05%,respectively.The quantity limit of ELISA method for quantitative detection of Anti-TT demonstrated to be 0.175 mIU/mL,its average recovery rate was 97.5%.The intra-assay CV and inter-assay CV of this Anti-TT assay were ≤ 2.42% and ≤5.58%,respectively.These two methods were applied for the immunogenicity evaluation after infantile basic immunization by diphtheria and tetanus vaccines.Conclusion The two established quantitative ELISA methods demonstrate high accuracy and good reproducibility,which are suitable for the ordinary laboratory to carry out the work and can be used in the serological effect evaluation after diphtheria and tetanus vaccine immunization and epidemiological study of diphtheria and tetanus disease.

Objective To develop an intelligent moxibustion instrument based on MCU,on the basis of moxibustion principles of traditional Chinese medicine.Methods A mixed-signal system-on-chip MCU C8051F020 was used to design four kinds of moxibustion heads to heat,light,and to upload detected temperature to the host microcontroller.Sixteen output ports were designed of which temperature and moxibustion time can be individually set.Results The temperature range of moxibustion head was 30-60℃,with error ≤±3 ℃.The timing range was 10-90 min,and full-scale timing error was ≤±1 min.The average infrared all normal emission rate of moxibustion head was 0.89.Conclusions The test parameters meet the requirements for moxibustion instrument,and the instrument is safe,reliable and simple to operate.

Background Permethrin is a commonly used pyrethroid insecticide and has been found to be potentially neurotoxic. Microglia are innate immune cells in the central nervous system and are involved in the development of a range of neurodegenerative diseases. Objective To observe possible toxic effects of permethrin on human microglia clone 3 (HMC3) in vitro and explore associated mechanism. Methods HMC3 were treated with 0, 10, 25, and 55 μmol·L⁻¹ permethrin for 72 h. Cell cycle and apoptosis were measured using flow cytometry. Cyclin-dependent kinase 1 (CDK1), cyclin-dependent kinase inhibitor 1A (CDKN1A), cyclin B2 (CCNB2), cellular tumor antigen p53 (p53), factor-related apoptosis (FAS), caspase 3 (CASP3), and H2A histone family member X (H2AX) were detected by quantitative real-time PCR (qPCR). The differential genes and enrichment pathways of HMC3 after 0 and 25 μmol·L⁻¹ permethrin treatment was analyzed by RNA sequencing. HMC3 was treated by 0, 10, 25, and 55 μmol· L⁻¹ permethrin for 72 h. The content of nitric oxide (NO) in the supernatant was detected using Griess reagent. The secretion level of interleukin-6 (IL-6) was detected by enzyme linked immunosorbent assay (ELISA). The mRNA expression levels of mitogen-activated protein kinase (MAPK) pathway (including MAPK1, MAPK8, and MAPK14), interleukin-1β (IL-1β), IL-6, and matrix metalloproteinase (MMP) families (including MMP1, MMP2, MMP3, and MMP9) were detected by qPCR. The protein expressions of phosphorylated p38 mitogen-activated protein kinase (p-p38), phosphorylated extracellular signal-regulated kinase (p-ERK), IL-1β, IL-6, and MMP1 were detected by Western blot. Results HMC3 was arrested in G2/M phase after 0, 10, 25, and 55 μmol·L⁻¹ permethrin treatment for 72 h, of which there was a statistically significant difference between the 55 μmol·L⁻¹ permethrin treatment group and the control group (P<0.01), and the mRNA expression of CDKN1A was up-regulated according to the qPCR (P<0.05). There was no statistically significant difference in the proportions of apoptosis between the groups (P＞0.05). The RNA sequencing showed that the differential genes were enriched in the MAPK pathway, and the mRNA expressions of MAPK1, MAPK8, and MAPK14 were up-regulated after the permethrin treatment at 55 μmol·L⁻¹ compared to the control group by qPCR (P<0.05). The Western blot revealed that, compared to the control group, the levels of p-p38 and p-ERK were increased after the 10 μmol·L⁻¹ permetrin treatment (P<0.05), the p-ERK level was increased after the 25 μmol·L⁻¹ permetrin treatment (P<0.05), and the p-p38 level was up-regulated after the 55 μmol·L⁻¹ permetrin treatment (P<0.05). The secretion of NO in the supernatant of HMC3 increased after permetrin treatment compared to the control group (P<0.05), the mRNA and protein expressions and the secretion of IL-6 showed an upward trend, the mRNA and protein expressions of IL-1β were up-regulated (P<0.05), and the mRNA and protein expressions of MMP1 were up-regulated in the 25 and 55 μmol·L⁻¹ permethrin groups (P<0.05). Conclusion Permethrin inhibits HMC3 cell proliferation in vitro, induces cell cycle arrest, activates MAPK pathway, and promotes the expression of inflammatory factors IL-1β and MMP1, which may be one of the mechanism of neurotoxicity induced by permethrin.

Objective To characterize the resistance mechanisms of a clinical Shigella sonnei strain harboring blaCTX-M-55 .Methods A double-disk synergy test was conducted to detect ESBL.Antibiotic resistance genes were determined by PCR followed by amplicon sequencing.Conjugation experiments were performed to verify the transferability of the plasmids carrying ESBL genes.The minimum inhibitory concentration values were tested using VITEK 2.The transposition unit was confirmed by DNA sequencer,and the transcriptional start site was identified using primer extension assay.Results Strain #1083 produced CTX-M-55,which was encoded by plasmid p1083-CTXM that could be transferred into E.coli through conjugation experiments to confer corresponding antibiotic resistance to the transconjugant #1083-EC600.The transposition unit mediating the transfer of blaCTX-M-55 was ISEcp1-blaCTX-M-55 -Δorf477.ISEcp1 offered strong promoter regions for the resistance genes,facilitating their expressions.Besides,the expressions were constant,not induced by antibiotics.Conclusion BlaCTX-M-55 on plasmids is the major resistance genes for strain #1083.Their expressions and spread are mediated by the insertion sequence ISEcp1.

Objective:To explore the clinical effect and application value of simultaneous modulated accelerated radiotherapy (SMART) in the suspicious positive lymph nodes of head and neck.Methods:From January 2017 to February 2019, 60 patients with suspected positive lymph nodes in the head and neck in the Hanzhong Central Hospital of Shaanxi Province were divided into experimental group and control group according to different treatment plans, and 30 patients in each group were included. In the experimental group, 63.36 to 66.66 Gy patients were treated with SMART, while in the control group, 54.12 to 60.06 Gy patients were treated with conventional neck prophylactic radiation. In order to evaluate the feasibility of the method, the change of the short diameter of the largest cross section of the suspicious positive lymph nodes in the two groups were observed, and the adverse reactions in the treatment of the two groups were analyzed.Results:There was no significant difference between the two groups before treatment ( P>0.05). After treatment, the size of short diameter of lymph nodes in the two groups was smaller than that before treatment. The maximum short diameter of the largest cross section of lymph nodes in the experimental group was smaller than that before treatment: (0.43 ± 0.07) cm vs. (0.72 ± 0.10) cm, and the difference was statistically significant ( P<0.05). In the control group, the maximum short diameter of the largest cross section of lymph node decreased after treatment, and the difference was not statistically significant ( P>0.05). After treatment, the reduction of the short diameter in the experimental group was more obvious than that in the control group. The maximum short diameter of the largest cross section between the two groups: (0.43±0.07) cm vs. (0.66±0.08)cm was statistically significant ( t = 11.523, P<0.05). Before treatment, hemoglobin (HGB) levels of the two groups were in the normal physiological range, and there was no significant difference between the two groups ( P>0.05); the white blood cell (WBC) levels of the two groups at different time after treatment were compared: in the first week (7.83 ± 2.53) × 10 9/L vs. (8.26 ± 3.16) × 10 9/L, in the third week (7.14 ± 3.65) × 10 9/L vs. (7.08 ± 2.53) × 10 9/L, in the fifth week (5.47 ± 2.81) × 10 9/L vs. (6.41 ± 2.57) × 10 9/L, and in the seventh week (4.36 ± 2.59) × 10 9/L vs. (4.98 ± 1.64) × 10 9/L, and there were statistical differences ( P<0.05), which indicated that the WBC index levels of the two groups were gradually decreased during the treatment, and the decreased degree of the experimental group was higher than that of the control group. The levels of HGB and PLT were maintained in the normal physiological range before and after treatment, and there was no significant difference between the two groups ( P>0.05). The main complications in the treatment of the experimental group were xerostomia and stomatitis. The adverse reactions in the control group were pain in the target area of radiotherapy. There was no significant difference between the two groups ( P>0.05). Conclusions:The application of IMRT is an effective method for the treatment of occult lymph node metastasis, and it is also a therapeutic diagnostic method, which can provide evidence for the study of the law of lymph node metastasis in the head and neck. The safety and tissue tolerance of IMRT in the treatment of suspicious positive lymph nodes in the head and neck are good, which can be used for the suspicious lymph nodes in the head and neck. The treatment of positive lymph nodes and the evaluation of patients′ prognosis provide an effective way of clinical treatment.

Objective:To construct talent training objectives and courses for undergraduate education of tropical medicine.Methods:Two rounds of questionnaire consultation were conducted among 15 experts by Delphi method. SPSS 26.0 software was used for statistical analysis, and the recovery rate, expert authority coefficient, mean of importance score, full score ratio, coefficient of variation and Kendall coordination coefficient were calculated respectively. Kendall's rank correlation test was used to analyze the degree of expert coordination, and the "boundary value method" was used to screen the indicators.Results:The effective recovery rates of the two rounds of consultation were all 100.00% and the expert authority coefficient was 0.815. The coordination coefficient was 0.25, 0.32, and 0.27, 0.36 respectively, and the significance test showed P<0.001. Finally, 11 talent training objectives and 7 courses for undergraduate education of tropical medicine were formed. Conclusions:The talent training objectives and courses for undergraduate education of tropical medicine are reasonable and reliable, which can provide theoretical support for tropical medicine talent training and have certain guiding value.