ABSTRACT:Objective To construct V-set and immunoglobulin domain containing 4 (Vsig4)nanobodies (Nbs) as specific macrophage probes so as to use them as molecular probes of macrophagocytes.Methods A nanobody phage library was generated by using peripheral blood lymphocytes isolated from an alpaca immunized with recombinant Vsig4 protein.After three rounds of selection against recombinant Vsig4.The Nbs were subjected to sequencing and genome alignment to obtain VHH sequence.Nbs were isolated and tested for Vsig4 specificity in an ELISA using recombinant Vsig4.The affinity capacity of Nbs was verified by the cell line stably expressing Vsig4. Results A nanobody phage library with an estimated 7.27 × 107 clones with 70% insertion was successfully constructed.Totally 1 3 6 Vsig4-positive clones were sequenced and aligned according to different CDR3 sequences. In summary,1 5 Vsig4 nanobodies were obtained and grouped into 3 different CDR3 epitopes.The affinity of representing nanobody and Vsig4 was analyzed via ELISA;Nb1 1 9 showed the highest affinity against both recombinant and native Vsig4.Conclusion We successfully constructed and screened Vsig4 specific nanobody number 1 1 9 with high affinity and specificity.It can help with macrophage detection and in vivo monitoring.

Objective To analyze the epidemiological features of leprosy in China from 2011 to 2015,and to provide scientific evidences for prevention and treatment strategies.Methods An epidemiological analysis and a trend analysis were conducted based on the national leprosy surveillance data from 2011 to 2015.Results The leprosy detection rate in China decreased from 0.085 per 100 000 in 2011 to 0.049 per 100 000 in 2015,with an average annual decline rate being 12.9％.A total of 4 775 leprosy cases were newly detected during 2011-2015,including 106 (2.2％) children,1 499 (31.4％) females,518 (10.8％) floating people,4 041 (84.6％) multibacillary cases and 1 134 (23.7％) cases with grade 2 disabilities.From 2011 to 2015,328 relapsed cases were reported,including 153 (46.6％) cases recurring after combined chemotherapy.The prevalence rate of leprosy in China decreased from 0.407 per 100 000 in 2011 to 0.235 per 100 000 in 2015,with an average annual decline rate being 12.9％.By the end of 2015,there had been 3 230 registered leprosy cases and 124 counties with a prevalence rate above 1 per 100 000.Conclusions The detection rate and prevalence rate of leprosy in China were both decreasing continuously from 2011 to 2015.The high-epidemic provinces were Yunnan,Guizhou,Sichuan and Guangdong.However,leprosy control in middle-and low-epidemic provinces can not be ignored.

Objective:To observed the effects of electroacupuncture frequency on the nerve function and postsynaptic destination-95 (PSD-95), growth associated protein-43 (GAP-43) in brain tissue of rats with cerebral ischemia were observed.Methods:Healthy adult male SD rats were randomly divided into the blank group, sham operation group, model group and 2 Hz, 50 Hz, 100 Hz electroacupuncture groups. MCAO models were prepared in each group, except blank group and sham operation group. After modeling, Qiansanli and Waiguan acupoints were given stimulation with 2 Hz, 50 Hz and 100 Hz respectively in 2 Hz electroacupuncture group, 50 Hz electroacupuncture group and 100 Hz electroacupuncture group, 1 time/d and 20 min/time for 21 consecutive days. All groups were scored by Bederson score on the 1st, 3rd, 7th, 14th and 21th day after operation. The ultrastructure of synapses was observed by transmission electron microscope, and the positive expression of PSD-95 and GAP-43 was detected by immunohistochemistry. Results:After modeling 7, 14 and 21 days, compared with the model group, the Bederson behavioral score (1.70 ± 0.52 vs. 2.40 ± 0.56, 1.20 ± 0.65 vs. 2.30 ± 0.46, 0.70 ± 0.72 vs. 2.00 ± 0.86) in 50 Hz electroacupuncture group significantly decreased ( P<0.01 or P<0.05). The expression of PSD-95 (58.67 ± 1.72, 55.22 ± 2.45, 60.10 ± 2.49 vs. 70.87 ± 2.34) intergral optical density of rats’ cortex in 2 Hz, 50 Hz, 100 Hz electroacupuncture groups significantly decreased ( P<0.01), the expression of GAP-43 in 2 Hz and 50 Hz electroacupuncture groups (58.89 ± 1.28, 64.76 ± 3.94 vs. 53.24 ± 2.58) significantly increased ( P<0.01). Conclusions:Electroacupuncture with different frequency can improve motor nerve disfunction of cerebral ischemia rats, and 50 Hz electroacupuncture group showed the most obvious effect. It can regulate PSD-95 and GAP-43 expressions of ischemic area, lower neural excitatory toxicity, increase neuronal axon regeneration and exert the neural protect function.

Objective This study was to understand the structure characteristics of prophages in the genome of Enterococcus hirae R17,and also to analyze their interaction relationships with the host bacterium.Methods The gene distribution and gene encoding characteristics of prophages in the genome of Enterococcus hirae R17 were identified using the PHAST software.The virulence gene,antimicrobial resistance genes,and environmental resistance genes in the prophages were also analyzed.Results Three prophages were found on the chromosome of Enterococcus hirae,including two incomplete prophage elements (Prophage-1 and Prophage-2) and one complete prophage (Prophage-3).Some function genes of bacteria were found in the sequence of three prophages,including nucleotide transportation and metabolism related genes.One incomplete prophage carrying erythromycin-and bacitracin-resistance genes was identified in the plasmid,which suggested that prophage induced gene horizontal transfer caused erythromycin-and bacitracin-resistance of Enterococcus hirae R17.Conclusion This study laid a solid foundation for the diversity analysis of prophages of Enterococcus hirae.Prophages played an important role in promotion of antimicrobial resistance of enterococci.Scientists should pay more attention to the spread of antimicrobial resistance and pathogenicity induced by prophages.

Objective To set up a three-dimensional finite element model (FEM) to investigate biomechanics of point contact locking plate (PC-LP) fixating femoral shaft fractures. Methods One intact fresh adult cadaveric femur was scanned by CT at 1 mm interval. Then, the data of CT were utilized to establish three-dimensional FEM by using software Mimics and PRO/E and simulate the different clini-cal loading conditions. The changes of theoretical stress of femur and PC-LP were analyzed under flexion, axial compression and torsion loads. Results (1) Under four-point bending load, the distribution of femur stress was in uniformity, with the largest stress of the PC-LP focused on the edge. (2) Under axial compression load of 250 N, the largest stress of the femur was focused on the screw holes on beth distal ends, with the largest stress of the PC-LP focused on the middle screw holes. (3) Under the torsion load cused on the middlepart and the middle screw holes. Conclusions Under the four-point bending, ax-ial compression and torsion loads, the distribution of femur stress is in uniformity, when the largest stress of the PC-LP focuses on edge or the middle screw holes, while that of the PC-LP on two screw holes of proximal or distal ends.

Objective To find out parental locus of control and mental health affecting students test anxiety. Methods The samples were from 5 middle schools including 1000 students,and their parents. They were investigated with the general life scales, Sarason's test anxiety scale, Symptom checklist 90 (SCL-90)and Parenting Locus of Control Scale(PLOC). Results The ratio of test anxiety: the mild was 28.8%, moderate was 45.0%, severe was 26.2% ,and felt anxiety was 55.8%. Ratio of test anxiety was higher in the boy students (X2=9.284, P =0.010) ,and heavier(F:16.42±6.70; M:15.18 ±6.51, t=2.716, P=0.007). Student test anxiety was significantly positively correlated with their parental mental health (r fatherl～11=0.166～0.272, Pfather l～11= 0.000;r mother 1～11=0.182～0.242, P mother 1～11= 0.000); Student test anxiety positively correlated with the effectiveness cof parents education, father's belief on the fate, eontrol of fathers for their children, locus of control from fathers (r1～5=0.075～0.143; P1-5=0.000～0.030);felt anxiety positively correlated with self-expect ,pressure from their mothers,over take care attitude from their parents,self-pressure from their parents( r1～6=0.068～0.230; P1-6=0.000～0.050) ,and negatively correlated with respect attitude of parents for their child, Marital relations,Communication time between the students and their parents,attitude of mother for learning( r1-7=-0.074～-0.140;P1～7=0.000～0.034). Felting test anxiety was served as a dependent factor, some factors enter the regression equation,and they were somatization of father, psychotic mother, the pressure from parents, mother over expectations, self-expectations ,the child control from student's father by Logistic regression analysis ( OR1～16=0.675～3.029;P1-6=0.000～0.007). Conclusion Students test anxiety is a common problem in male and female students. Mental health and locus of control from students' parents show test anxiety has somatization of father, psychotic mother, the pressure from parents, mother' expectations,self-expectations, the control of father for the child 6 risk factors.

Objective: To evaluate health risks of five heavy metals in drinking water of rural schools in Henan Province, so as to provide scientific basis for drinking water safety in rural schools. Methods: Totally 1 269 drinking water samples were collected and analyzed for five heavy metals (As, Cd, Cr 6+ , Pb, Hg) concentration in 106 cities and counties of Henan Province, and its spatial distribution characteristics were explored by geographic information system (GIS). Risk assessment method recommended by the United States Environmental Protection Agency (U.S. EPA) was applied to evaluate health risks from five heavy metals through oral ingestion. Results: The qualified rate of As concentration was 100%, and the qualified rates of Cd, Cr 6+ , Pb and Hg were 99.9% , 99.9%, 99.5%, 99.6%. The highest carcinogenic risk was As (3.05×10 -5 ), followed by Cr 6+ (2.73×10 -5 ), and the highest non carcinogenic risk was As (0.158 3), followed by Pb(0.041 7). The carcinogenic risk and non carcinogenic risk were all higher in girls than boys. The carcinogenic and non carcinogenic risks were differences in different regions( P <0.05). Conclusion Health risks of five heavy metals in drinking water for rural schools in Henan Province were within the acceptable risk level recommended by EPA. As, Cr 6+ and Pb were the main health risk factors in drinking water, and that might impact negatively the healthy growth of primary and middle school students. These should be future efforts of drinking water safety management.

Objective: To study the effects of atmospheric fine particulate pollution on the lung function of primary school students before and after heating during the winter in Zhengzhou. Methods: In Zhengzhou, two areas with low and high level of PM 2.5 pollution(A and B), were selected as monitoring points from 2016 to 2018. Each monitoring station selected one elementary school within around 1 km and used a cluster random sampling method to extract students from grades 3 to 5 as the research subjects. Lung function tests were conducted before and after heating in winter, and mass concentration of PM 2.5 were recorded daily and compared to those recorded one month prior. Results: The average daily mass concentration of PM 2.5 were 74 μg/m 3 and 92 μg/m 3 in the light pollution monitoring points A and the heavy pollution monitoring points B, which exceeded the standard for 97 and 126 days, respectively. The FVC and FEV1.0 indexes for the first test of male students before heating were higher than those for the second test from 2016-2018 (P<0.05). Except in 2016,the FVC and FEV1.0 indexes for the first test of female students were also higher than those for the second test (P<0.05). After stratified by sex,multivariate linear regression showed that PM 2.5 was associated with lung function as measured by the FEV1.0, PEF, FEF25 and FEF75 values of the students were negatively correlated (B=-0.13--0.07,-0.13--0.08,P<0.05). Conclusion Fine particulate air pollution before and after heating during the winter in Zhengzhou had different effects on the lung function of primary school students. Therefore, it is necessary to strengthen the respiratory health protection of primary school students in winter to protect their health.

ObjectiveTo explore the influencing factors of occupational hand-arm vibration disease (OHAVD) caused by handheld workpiece polishing. Methods A total of 222 OHAVD patients (case group), 275 hand-transmitted vibration-exposed workers (exposed group) and 243 healthy workers without hand-transmitted vibration exposure (control group) in a sports equipment manufacturing enterprise were selected as the study subjects using the convenience sampling method. Worksite survey of occupational health was conducted on these three groups, and the human vibration measurement equipment was used to measure the vibration exposure level of handheld vibration among the study subjects. The 8-hour energy equivalent frequency-weighted vibrating acceleration ［A(8)］ and cumulative vibration exposure level (CVEL) were calculated. Results The prevalence of coldness, numbness, tingling fingers, and vibration-induced white finger was higher in the exposed group and the case group compared with the control group (all P<0.05). The prevalence of the above-mentioned hand symptoms was higher in the case group compared with the exposed group (all P<0.05). The A(8) and CVEL levels of the study subjects in the case group were higher than those in the exposed group (all P<0.05). Binary logistic analysis result showed that age and CVEL were both influencing factors of OHAVD (all P<0.05). According to the restricted cubic spline models, CVEL of the study subjects in the exposed group had a positive nonlinear dose-response relationship with the risk of OHAVD (overall trend P<0.01, nonlinear P<0.01), indicating an increasing risk of OHAVD with increasing CVEL. Conclusion Hand-transmitted vibration exposure is a risk factor for OHAVD. Early intervention should be carried out for hand-transmitted vibration-exposed individuals to reduce vibration-exposed levels and control vibration exposure time.

Objective:To select human chronic myeloid leukemia (CML) cell line K562 as the experimental object, and use lentivirus mediated CRISPR/Cas9 gene editing technology to construct a stable CML cell line K562/TCRP1-KO that knocks out the tongue cancer resistance related protein 1 (TCRP1) gene; and through functional tests such as cell proliferation, apoptosis, and drug sensitivity, compare the phenotypic differences between K562/TCRP1-KO and control cells (K562/cas9-CTL), and preliminarily explore the possible mechanism of TCRP1 gene involvement in the pathogenesis of CML.Methods:The small guide RNA (sgRNA) targeting TCRP1 was designed at a specific location. After annealing, the oligonucleotide fragments were recombined with the linearized Cas9 expression vector, and the lentivirus packaging system was transfected into 293T cells. The purified virus was collected and infected with K562 cells. Positive polyclons were screened for puromycin pressure, and monoclonal K562/TCRP1-KO was further screened by limited dilution method. Stable cell lines were successfully knocked out by sanger sequencing and Western blot detection; Simultaneously, K562 cells transfected with lentiCRISPR vector were constructed as control cell lines (K562/cas9-CTL); Using cell counting method, cell counting kit 8 (CCK8) method, imatinib (IM) gradient dilution method, and flow cytometry cell proliferation, drug sensitivity, and apoptosis analysis were performed on K562/TCRP1-KO and K562/cas9-CTL, respectively.Results:The sgRNA-Cas9 recombinant plasmid vector for TCRP1 knockout was successfully constructed, and after transfection into 293T cells, TCRP1 knockout monoclonal cell lines were successfully screened using limited dilution method. Compared with K562/cas9-CTL cells, the proliferation ability of K562/TCRP1-KO cells was significantly reduced, IM drug sensitivity was significantly enhanced, and the process of cell apoptosis was significantly accelerated (all P<0.05). Conclusions:A CML cell line with TCRP1 knockout was successfully constructed using CRISPR/Cas9. TCRP1 may act as a cancer related gene to affect the proliferation, IM resistance, and apoptosis process of CML cells.