Abstract: Objective To construct a shuttle vector pHT315-AaCPR100A with two spore-producing-dependent promoters and the target gene AaCPR100A in Escherichia coli-Bacillus thuringiensis. Methods The forward promoter of Cry3A, named Pro-1 (+), was amplified by PCR using pSVP27A plasmid as the template, and the target gene AaCPR100A was amplified using Aedes aegypti RNA reverse conversion cDNA as the template. The plasmid pHT315 was linearized by digestion with Hind Ⅲ and Sal Ⅰ. The forward promoter and the target gene were inserted into the linearized vector pHT315 successively by in-fusion cloning according to the transcription direction. The synthesized plasmid containing the Cry3A reverse promoter sequence was used as the template, and the Pro-1 (-) reverse promoter was amplified by PCR. The intermediate vector containing the forward promoter and the target gene was linearized by EcoR I restriction enzyme, and the reverse promoter was inserted downstream of the target gene by in-fusion cloning in the direction of transcription. Results By agarose gel electrophoresis, the forward promoter, target gene AaCPR100A and reverse promoter bands were clear and of good quality, which could be used for in-fusion cloning experiments. The two spore-producing-dependent promoters and target gene fragments were connected by In-fusion cloning. The recombinant vector pHT315-AaCPR100A was verified by PCR. The forward promoter, target gene fragment and reverse promoter were successfully amplified in the recombinant vector. Nucleotide sequencing verified that the sequencing results of the bidirectional promoter sequence and the target gene sequence were basically consistent with the sequence alignment results, which met the requirements of the construction of vector elements and proved that the recombinant vector was successfully constructed. Conclusions Based on the above results, this study proves that the recombinant shuttle vector with two spore-producing-dependent promoters can be successfully constructed by in-fusion cloning technology, laying the foundation for the construction of engineered Bacillus thuringiensis expressing dsRNA of AaCPR100A.

Objective To investigate the relationship and mechanism of the serum vitamin D levels with insulin sensitivity and adipokines APN,RBP4,visfatin and TNF-α in patients with gestational diabetes mellitus(GDM). Methods A total of 101 patients with gestational diabetes( GDM group) and 50 pregnant women with normal blood sugar( control group) were recruited to detect the serum levels of correlative biochemical indexes,APN,RBP4,visfatin,TNF-α and 25OHD3 using an HPLC method. Twenty-four patients in the GDM group diagnosed with vitamin D deficiency(<25 nmol·L-1 ) were randomized to receive either 1,25(OH) 2D3(0. 25 μg·d-1) orally or no treatment. Serum levels of correlative biochemical indexes were detected after 1 month. Results GDM patients had 25OHD3 levels significantly lower than the control group(P<0. 05),APN levels lower than the control group,and RBP4,visfatin,TNF-α levels significantly higher than the control group(all P<0. 05). After 1 month of 1,25(OH) 2D3 treatment,HOMA-IR increased,while HOMA-β decreased. APN levels was positively correlated(r= 0. 526) with 25OHD3;RBP4,visfatin,TNF-α decreased and were negatively correlated with 25OHD3( r values were -0. 272,-0. 153,-0. 072). Conclusion Vitamin D can reduce the adipokines RBP4,visfatin,TNF-α,increase APN,and thus play a protective role for gestational diabetes.

Objective Currently , the targeted therapy is the first-choice treatment of advanced non-small cell lung cancer (NSCLC) in with epidermal growth factor receptor (EGFR) mutations, but few studies have been reported on the relationship between immunohistochemical markers and the EGFR mutation.The aim of this study is to analyze the relationship of the EGFR mutation with the ex-pressions of thymidylate synthetase (TS), excision repair cross-com-plementation group 1 ( ERCC1 ) , β-tubulin-III, and ribonucleofide reductase large subunit-l ( RRM1) in NSCLC. Methods We retro-spectively analyzed 336 cases of NSCLC treated in the Department of Medical Oncology , the Affiliated Hospital of Inner Mongolia Medical University, from June 2014 to December 2015 and examined 29 EGFR mutations.We divided the patients into a mutation and a non-mutation group, performed immunohistochemical staining of the TS, ERCC1,β-tubulin-III and RRM1 proteins and compared their expressions in the NSCLC tissue between the two groups . Results EGFR mutations were found in 138 ( 41.07%) of the 336 NSCLC patients but not in the other 198 ( 58.93%) .The expression of TS was significantly lower in the mutation than in the non-mutation group (9.42%vs 39.39%, P<0.05), and so was that of β-tubulin-III (44.2%vs 60.1%, P<0.05).EGFR mutations were correlated with decreased expressions of TS (r=-0.332, P<0.05) andβ-tubulin-III (r=-0.157, P<0.05).Multivariate regression analysis showed that the risk of EGFR mutations was 2.109 times higher in the fe-male patients than in the males (OR=2.109, 95%CI:1.268-3.509), 24.265 times higher in the adenocarcinoma than in the adeno-squamous carcinoma patients (OR=24.265, 95%CI:3.508-167.845), 15.2 times higher in the squamous carcinoma than in the ade-nocarcinoma patients (OR=15.200, 95%CI:4.480-51.569), 2.364 times higher in the lung biopsy specimens than in the surgically treated patients (OR=2.364, 95%CI:1.266-4.413), and 6.171 times higher in the patients with lowly expressed than in those with highly expressed TS (OR=6.171, 95%CI: 3.145-12.109). Conclusion The decreased expressions of TS and β-tubulin-Ⅲ in NSCLC indicate the mutation of the EGFR gene.

The paper analyzes the cause for poor medication compliance of hypertensive patients,indicates the necessity of development of hypertensive health management APP,designs three core functions of the APP including health science popularization,medication reminding and blood pressure monitoring,promotes the improvement of medication compliance of patients,and improves the use intensity of users of the APP through the internal incentive measures that combining health belief and effect monitoring and the external incentive measures that focus on the reward system.

In order to understand the situation of schistosomiasis in Fujian Province, the surveillance and field survey data of 16 counties (cities or districts) were statistically analyzed from 2003 to 2008 according to the national and provincial surveillance protocols. The snail areas were 69 400 m~2 in 2003, and 506 700 m~2 in 2004, then decreased to 57 700 m~2 in 2008 after the snail control in 2005 and 2006. Serological examinations were conducted in 18 000 people, among which 5 412 people were the mobile population from schistosome-endemic areas, and 110 persons were positive, among which 8 persons had eggs and miracidia in their stools, among 7 885 appendix samples, Schistosoma japonicum eggs were found in 8 samples of the patients who came from the o-riginal endemic areas. By the stool examination, 2 709 head of farm cattle in the region with snails were not found infected with Schistosoma japonicum. It is suggested that the areas of residual snails are large and widespread, and the imported patients are found frequently, so the danger of schistosomisis transmission still remains.

Objective To explore the change patterns in the lumbar vertebral bone marrow fat content with age, analyze the interactions between lumbar vertebral bone marrow fat content, bone mineral density (BMD), and age, and compare the difference of lumbar vertebral bone marrow fat content between males and females in the same age groups. Methods According to the statistical sample size requirements that the minimum sample size of each group was 18, thus, we prospectively recruited healthy volunteers who met the inclusion criteria. Among them, there were 139 males and females, with age range of 21-70 years old. According to age, the patients were divided into group 1 (21-30-yr), group 2 (31-40-yr), group 3 (41-50-yr), group 4 (51-60-yr), and group 5 (61-70-yr). Quantitative computed tomography (QCT) was used to measure the lumbar vertebral BMD, and MR mDIXON-Quant technique was used to measure the marrow fat content of L3 lumbar vertebra. We compared the difference of lumbar vertebral bone marrow fat content and BMD between different age groups in males and females using one-way ANOVA, and compared the difference of lumbar vertebral bone marrow fat content and BMD between males and females in the same age groups using t-test. Correlation analysis was conducted between bone marrow fat content, BMD and age. Results Lumbar vertebral bone marrow fat content was generally increasing with the age. There were significant differences in the fat content of bone marrow at different age groups (male, F=13.598, P=0.000;female, F=73.419, P=0.000). Before the age of 50 years, lumbar vertebral bone marrow fat content in females was lower than one in males, and there was a significant difference between group 2 [females,(29.7± 7.1)%-(36.1 ± 6.6)%, males,(34.1 ± 8.4)%-(39.9 ± 5.9)%;21-30-yr, t=1.984, P=0.053;31-40-yr, t=5.699, P=0.000;41-50-yr, t=2.017, P=0.050]. Females older than 50 years had a higher marrow fat content than males, and there was a significant difference between group 5 [females,(48.3±8.8)%-(52.5±8.2)%, males, (45.5 ± 8.1)%-(46.2 ± 7.4)%;51-60-yr, t=-0.914, P=0.365;61-70-yr, t=-3.400, P=0.001]. For males, bone marrow fat content was positively correlated with age (r=0.527, P<0.05), and negatively correlated with BMD (r=-0.730, adjusted for age r=-0.584, P<0.05). For females, bone marrow fat content was positively correlated with age (r=0.761, P<0.05), and negatively correlated with BMD (r=-0.809, adjusted for age r=-0.473, P<0.05). Conclusions Lumbar vertebral bone marrow fat content was generally increasing with the age. Bone marrow fat content was positively correlated with age and negatively correlated with BMD for males and females.

Objective To explore the change patterns in the lumbar vertebral bone marrow fat content with age, analyze the interactions between lumbar vertebral bone marrow fat content, bone mineral density (BMD), and age, and compare the difference of lumbar vertebral bone marrow fat content between males and females in the same age groups. Methods According to the statistical sample size requirements that the minimum sample size of each group was 18, thus, we prospectively recruited healthy volunteers who met the inclusion criteria. Among them, there were 139 males and females, with age range of 21-70 years old. According to age, the patients were divided into group 1 (21-30-yr), group 2 (31-40-yr), group 3 (41-50-yr), group 4 (51-60-yr), and group 5 (61-70-yr). Quantitative computed tomography (QCT) was used to measure the lumbar vertebral BMD, and MR mDIXON-Quant technique was used to measure the marrow fat content of L3 lumbar vertebra. We compared the difference of lumbar vertebral bone marrow fat content and BMD between different age groups in males and females using one-way ANOVA, and compared the difference of lumbar vertebral bone marrow fat content and BMD between males and females in the same age groups using t-test. Correlation analysis was conducted between bone marrow fat content, BMD and age. Results Lumbar vertebral bone marrow fat content was generally increasing with the age. There were significant differences in the fat content of bone marrow at different age groups (male, F=13.598, P=0.000;female, F=73.419, P=0.000). Before the age of 50 years, lumbar vertebral bone marrow fat content in females was lower than one in males, and there was a significant difference between group 2 [females,(29.7± 7.1)%-(36.1 ± 6.6)%, males,(34.1 ± 8.4)%-(39.9 ± 5.9)%;21-30-yr, t=1.984, P=0.053;31-40-yr, t=5.699, P=0.000;41-50-yr, t=2.017, P=0.050]. Females older than 50 years had a higher marrow fat content than males, and there was a significant difference between group 5 [females,(48.3±8.8)%-(52.5±8.2)%, males, (45.5 ± 8.1)%-(46.2 ± 7.4)%;51-60-yr, t=-0.914, P=0.365;61-70-yr, t=-3.400, P=0.001]. For males, bone marrow fat content was positively correlated with age (r=0.527, P<0.05), and negatively correlated with BMD (r=-0.730, adjusted for age r=-0.584, P<0.05). For females, bone marrow fat content was positively correlated with age (r=0.761, P<0.05), and negatively correlated with BMD (r=-0.809, adjusted for age r=-0.473, P<0.05). Conclusions Lumbar vertebral bone marrow fat content was generally increasing with the age. Bone marrow fat content was positively correlated with age and negatively correlated with BMD for males and females.

OBJECTIVE: To investigate the bone mineral density (BMD) of cervical vertebrae in a population-stratified manner and correlate with that of the lumbar vertebrae. MATERIALS AND METHODS: Five hundred and ninety-eight healthy volunteers (254 males, 344 females), ranging from 20 to 64 years of age, were recruited for volumetric BMD (vBMD) measurements by quantitative computed tomography. Basic information (age, height, weight, waistline, and hipline), and vBMD of the cervical and lumbar vertebrae (C2-7 and L2-4) were recorded. Comparisons among sex, age groups and different levels of vertebrae were analyzed using analysis of variance. Linear regression was performed for relevance of different vertebral levels. RESULTS: The vBMD of cervical and lumbar vertebrae was higher in females than males in each age group. The vBMD of the cervical and lumbar vertebrae in males and the vBMD of lumbar vertebrae in females decreased with aging. In each age group, the vBMD of the cervical vertebrae was higher than that of the lumbar vertebrae with gradual decreases from C2 to C7 except for C3; moreover, the vBMD of C6 and C7 was significantly different from that of C2-5. Correlations of vBMD among different cervical vertebrae (females: r = 0.62-0.94; males: r = 0.63-0.94) and lumbar vertebrae (males: r = 0.93-0.98; females: r = 0.82-0.97) were statistically significant at each age group. CONCLUSION: The present study provided normative data of cervical vertebrae in an age- and sex-stratified manner. Sex differences in vBMD prominently vary with age, which can be helpful to design a more comprehensive pre-operative surgical plan.
Aging ; Asian Continental Ancestry Group* ; Bone Density* ; Cervical Vertebrae ; Female ; Healthy Volunteers ; Humans ; Linear Models ; Lumbar Vertebrae* ; Male ; Sex Characteristics ; Spine

Abdominoscrotal hydrocele is rare in clinic. In the past, routine examination and diagnosis were difficult, easy to be misdiagnosed. The daily operations were mostly completed through the groin area or abdominal incision, the wound is large. The application of laparoscopy can clearly diagnose the abdominoscrotal hydrocele through "springing back ball" sign, and can cure the disease by laparoscopic resection of interperitoneal mass and closure of the internal ring. It is worthy of clinical application. In this article, we summarized and analyzed the clinical experience of 15 cases of children with abdominoscrotal hydrocele diagnosed and treated by laparoscopy, to explore the value of the laparoscopic technology in the diagnosis and treatment of the abdominoscrotal hydrocele.

Objective To investigate the influence of abdominal adipose tissue on insulin resistance and other metabolic indicators in the patients with type 2 diabetes mellitus(T2DM)complicating obesity.Methods Fifty-one patients with newly diagnosed T2DM complicating obesity (T2DM group) and 50 cases of simple obesity(control group) in Jishuitan Hospital were enrolled.The two groups were performed QCT for measuring the total abdominal adipose tissue volume (TAV),subcutaneous adipose tissue volume (SAV),visceral adipose tissue volume (VAV),and then the VAV/SAV ratio was calculated.Meanwhile the waist circumference,body mass index(BMI),blood uric acid (UA),total cholesterol (C H O),triglycerides (TG),high-density lipo protein cholesterol (HDL),low density lipoprotein cholesterol(LDL),homocysteine(HCY),dosage of basal insulin(DBIns),dosage of prandial insulin (DPIns),fasting plasma glucose(FBG),fasting blood insulin (FIns) and HbA1c levels were also measured.The relationship between TAV,SAV and VAV with HOMA-RI and other indicators was investigated.The influence of abdominal adipose tissue on insulin resistance and other metabolic indicators was determined.Results The UA,TG,LDL,FPG and HbA1c levels in the T2DM group were increased compared with the control group(P＜0.05).Compared with the control group,the SAV level in the T2DM group was decreased(P＜0.05),the VAV and VAV/SAV levels were increased(P＜0.05).The multivariate regression analysis showed that VAV was the natural logarithm affecting In (HOMA-RI),UA,CHO,TG,LDL,HCY,DBIns (β=0.399,0.398,0.389,0.447,0.440,0.372,0.307;P＜ 0.05).TAV was the main factor affecting the waist circumference and BMI (β=0.868,0.639;P＜0.05).Conclusion The visceral adipose tissue volume has larger influences on insulin resistance,blood lipids,UA,HCY and DBIns.