Porphyromonas gingivalis is a specific causative agent of human chronic periodontitis. This anaerobe produce collagenase PrtC encoded by prtC. To constructed the prokaryotic expression system for the prtC gene from P.gingivalis so as to be used to explore antigenicity and immunoreactivity of prtC gene product as well as the relationship between the local level of PrtC and the periodontitis damage, the entire length of prtC genes fragment from the ATCC-33277 and 47A-1 strains of P.gingivalis was amplified by PCR. After T-A cloning and sequencing, the prokaryotic expression system for prtC was constructed by using pET32a plasmid and E.coli BL21DE3 strain. The expression of the target recombinant PrtC protein was induce with different concentrations of IPTG. Western blot assay was used to detect the antigenicity and immunoreactivity of PrtC protein, and ELISA assay was used to detect the PrtC level in the subgingival plaque samples of patients with chronic periodontitis. The experimental results showed that the nucleotide sequences of the prtC genes from ATCC-33277 and 47A-1 strains of P.gingivalis were entirely identical, and the sequence similarities of nucleotides and amino acids were 98.46% and 99.07% respectively. Under the induction of different concentration of IPTG, the output of recombinant expressed product PrtC may reach up to 50% of the total bacterial proteins. It was also proved that the recombinant PrtC could bind with antibody against the whole cell of P.gingivalis, and could induce the production of specific antibodies in rabbit. In 91.39% of the subgingival plaque samples, the PrtC could be detectable, in which the level of positive rates of detection was higher in severe cases of chronic periodontitis than that in the mild cases. So far, a prokaryotic expression system for the prtC gene from P.gingivalis with high expression efficiency was successfully constructed in the present study, and the expressed product PrtC possesses well antigenicity and immunoreactivity, suggesting the possibility to be used as the candidate antigen for developing the serological kit and P.gingivalis vaccine.

Objective To investigate the correlation of anthropometric indicators and blood pressure in children. Methods A total of 6 790 children aged 6 to 13 years were inspected by random sampling. The body mass index (BMI), waist circumference (WC), hip circumference (HC), and blood pressure were measured. The waist/hip ratio (WHR) and waist/height ratio (WHtR) were calculated. The data wrer analyzed by SPSS16.0 statistical software. Results The detection rate of hypertension in children was 5.57%. After controlling for age, both in male and female children, the BMI, WC, HC, WHR, WHtR and systolic blood pressure showed a significant positive correlation by partial correlation analysis (all P<0.05). Both in male and female children, the BMI, WC, HC, WHR, and WHtR were significantly higher in children with hypertension than those in children with normal blood pressure (all P<0.05). Among all subjects, 280 children (4.12%) were obese, 622 children (9.16%) were overweight. The detection rates of hy-pertension were significantly different among obese, overweight and normal weight children (P<0.01). The detection rate of hypertension was significantly higher in obese children than that in overweight and normal weight children. The systolic and diastolic blood pressure were all significantly higher in obese and overweight children than that in normal weight children (P<0.05). Conclusion The hypertension prevalence of children aged 6 to 13 years in Zhengzhou is in the low to median level in the same age groups. The BMI, WC, HC, WHR, WHtR are signiifcantly correlated with blood pressure in both gender, and the correlation is the most obvious in HC.

Objective Boy sexual development is affected not only by age, but also by environment factors .The present study was to survey and evaluate the status of sexual development in boys from Zhengzhou area and to explore their influencing factors. Methods The height, weight, waist circumference, hip circumference, penis and testicles in 3779 boys aged from 6 to 13 years old in Zhengzhou were measured .The penis length and testicular volume of different ages and their influencing factors were compared, and the beginning age and sexual development characteristics were explored .The development of genitalia and pubes were evaluated by Tanner staging criteria and standards, which were divided into five stages. Results There were 3779 boys, including 395 overweight cases (10.45%) and 208 obesity cases (5.50%).The testicular volume and penis length increased by age, which increased slowly at the age of 6 to 11, followed by a rapid increase after the age of 11 year old.Testicular volume increased more rapidly than that of penis length. The medians quartile of testicular volume in overweight group ,obese group and normal group were 2.50 mL, 2.50 mL, 2.00 mL, 2.00 mL,2.00 mL,1.00 mL respectively .The testicular volume in overweight group and obese group were significantly greater than that in nor -mal group (P<0.01) and there were no difference between overweight group and obese group .The penis length in overweight group and obese group were significantly less than that in normal group (P<0.01), and there was no difference between overweight group and obese group.Testicular volume and penis length were positively correlated with age, height, weight, body mass index, waist circumference and hip circumference; the correlation between testicular volume and height, penis length and age were the strongest. The age of development of testicle (average 12.02 years) in urban area was later by 2.73 years than that in the rural area (average 9.9 years), the difference was statistically significant (P<0.001).The age of pubic development (P2) and maturity (P5) was (12.34 ±0.83) years and (12.39 ±1.24 ) years.The incidence of precocity in boys was 0.40%. Conclusion The sexual development of boys from Zheng-zhou area is similar to the results of large scale survey from China .Attention should be paid to the effect of obesity on sexual develop-ment, and early intervention should be given, the occurrence of adverse events of sexual development can be avoided .

Objective To summarize the correlation between cytotoxic T lymphocyte associated antigen-4 (CTLA-4) promoter-49 A/G polymorphism and Graves' disease (GD)in China.Methods The publications before Sep.30,2011 about the correlation between CTLA-4 promoter-49 A/G polymorphism and GD in China were collected by searching CNKI,CBM,and Wanfang database.Meta-analysis was performed to check the difference of genotypes such as AG,GG,GG + AG,and G allele between GD group and the control group.Results A total of 6 studies involving 570 cases in GD group and 486 cases in the control group were included in the meta-analysis.According to the heterogeneity of these studies,meta-analysis was performed to analyze the correlation between AG,GG,GG + AG genotype,G allele and GD by fixed effects model.The pooled OR value(95％ CI) of various studies was 1.97 (1.37-2.82),3.66 (2.42-5.53),2.55 (1.82-3.58),and 1.99 (1.49-2.65).Conclusion The meta-analysis demonstrates that CTLA-4 promoter-49 A/G polymorphism increases the possibility of getting GD in Chinese.

Objective To explore the relationship between the expression of XRCC1 and glioma. Methods Total of 26 samples of glioma were divided into 4 groups:gradeⅠ,gradeⅡ,gradeⅢand gradeⅣ. The expression of XRCC1 in 26 Gliomas tissues were examined using SP immunohistochemical staining.Results The positive staining of XRCC1 protein was localized in nucleus of tumor cells in Glioma. There was no correlation among them. The difference of XRCC1 expression among gradeⅠ~Ⅳ was not significant ( >0.05) .Conclusion The difference of XRCC1 expression among gradeⅠ~Ⅳ was not significant. The expression of XRCC1 was closely correlated with the effect of radiotherapy.

Objective To investigate the molecular mechanism of retinoic acid in targeted treatment of craniopharyngioma by detecting the expression of RARαand PPARβ/δin craniopharyngioma cells and analyzing the correlation between the expression and effect of retinoic acid. Methods The expression of RARα and PPARβ/δ in craniopharyngioma cells from 31 patients cultured in vitro was quantified by reverse transcription-PCR. The inhibition rates of RA on craniopharyngioma with different expression of RARα and PPARβ/δ were detected by using MTT assay, and the correlation between the expression of RARα and PPARβ/δand the effect of RA was analyzed. Results 1. The RT-PCR results showed that the expression levels of PPARβ/δand RARα mRNA were different. Craniopharyngioma cells from 31 patients in primary culture were divided into three groups according the expression levels of nuclear receptor: PPARβ/δ>RARα group, RARα>PPARβ/δ group and RARα>>PPARβ/δ group. 2.MTT results showed that the inhibition rate of RARα>>PPARβ/δgroup was significantly higher than the other groups under same drug, the differences had statistical significance ( <0.01) . Conclusions The expression of PPARβ/δ, RARα can be used to evaluate the effect of RA in treatment of craniopharyngioma. The craniopharyngioma with low-expression of PPARβ/δ is more sensitive to RA. Targeting higher RARα or targeting lower PPARβ/δ is beneficial to the treatment of craniopharyngiomas.

Objective:To analyze the characteristics of clinical manifestation, auxiliary examination and gene mutation of 3-hydroxy-isobutyryl-coenzyme A hydrolase (HIBCH) deficiency to better understand this disease.Methods:The clinical manifestations and genetic results of a patient with HIBCH deficiency were analyzed. The clinical features and genetic characteristics of HIBCH deficiency were summarized based on the literature review.Results:The proband, female, one year and four months old, was admitted to Children′s Hospital Affiliated to Zhengzhou University for “vomiting and diarrhea for 15 days, dyspnea and intermittent convulsions for 13 days after digestive tract infection”. The intelligence was normal, however, the motor development was slightly delayed before onset. Physical examination showed light coma, poor response and insensitivity to light. She also had shortness of breath, weak positive three concave signs and coarse breath sound in both lungs with sputum purrs. In addition, the muscle tension of extremities was increased. Bilateral Brudzinski′s sign, Babinski′s sign and Kernig′s sign were negative. Serum hydroxybutyryl carnitine (C4OH) was increased. Cranial magnetic resonance imaging (MRI) showed atrophy in bilateral cerebral hemispheres and abnormal symmetry signals in bilateral globus pallidus and cerebral peduncle. Novel compound heterozygous variants of HIBCH, c.489T>A (p. C163*) and c.740A>G (p. Y247C), were found in the patient, which respectively inherited from her healthy parents. Her symptoms were relieved after“cocktail”therapy and symptomatic treatment. Literature related to HIBCH deficiency published all around the world was reviewed. As a result, 17 articles, including 24 cases, had been reported. The majority of patients presented with poor feeding, dystonia and progressive motor developmental delay in early infancy. Cranial MRI showed lesions in bilateral basal ganglia. Serum C4OH concentration was elevated. And compound heterozygous or homozygous variants of HIBCH gene were found in patients with HIBCH deficiency.Conclusions:The detection of serum amino acids and acylcarnitine profiles on HIBCH deficiency was relatively specific and it was helpful to make a clear diagnosis by combining with cranial MRI and genetic tests. In this study, a case of HIBCH deficiency was confirmed, which expanded the mutation spectrum of HIBCH gene. Meanwhile, summarizing the clinical and genetic characteristics of cases reported improved understanding of HIBCH deficiency.

To analyze the types of dietary patterns of women during pregnancy in Shaanxi and their association with adverse pregnancy outcomes. Methods Data were derived from a cross?sectional program named "The prevalence and risk factors of birth defects in Shaanxi Province" from July to November in 2013. A multi?stage stratified random sampling method was used to extract 15 980 women of childbearing age who met the inclusion exclusion criteria from Shaanxi Province. The questionnaire obtained information such as the pregnancy outcome and the food intake of the study subjects during pregnancy. The dietary pattern was extracted by factor analysis, and the each dietary pattern of subjects were divided into T1-T3 groups according to the factor score. The effects of each dietary pattern on major adverse pregnancy outcomes were estimated by unconditional logistic regression model. Results Four dietary patterns were established: vegetarian pattern, balanced pattern, traditional pattern, and processing pattern. The multivariate logistic regression model results showed that compared with the vegetarian pattern T2 group, the T1 group had lower risk of low birth weight (OR=0.56, 95%CI : 0.41-0.83), and the T 3 group had higher risk of low birth weight in offspring ( OR=2.32, 95%CI : 1.59-3.89); compared with the traditional pattern T2 group, the T3 group had higher risk of premature (OR=2.62, 95%CI :1.58-5.01); compared with the balanced pattern T2 group, the T3 group had a lower risk of spontaneous abortion (OR=0.73, 95%CI: 0.36-0.89); compared with the processing pattern T2 group, the T3 group had a higher risk of spontaneous abortion ( OR=1.97, 95%CI : 1.36-3.34) and higher risk of stillbirth ( OR=2.96, 95%CI : 1.49-6.26), and the T 1 group had a lower risk of stillbirths in offspring ( OR=0.52, 95%CI :0.33-0.83). Conclusion The women of childbearing age in Shaanxi have different dietary patterns, and there may be a correlation between dietary patterns and adverse pregnancy outcomes.

To analyze the types of dietary patterns of women during pregnancy in Shaanxi and their association with adverse pregnancy outcomes. Methods Data were derived from a cross?sectional program named "The prevalence and risk factors of birth defects in Shaanxi Province" from July to November in 2013. A multi?stage stratified random sampling method was used to extract 15 980 women of childbearing age who met the inclusion exclusion criteria from Shaanxi Province. The questionnaire obtained information such as the pregnancy outcome and the food intake of the study subjects during pregnancy. The dietary pattern was extracted by factor analysis, and the each dietary pattern of subjects were divided into T1-T3 groups according to the factor score. The effects of each dietary pattern on major adverse pregnancy outcomes were estimated by unconditional logistic regression model. Results Four dietary patterns were established: vegetarian pattern, balanced pattern, traditional pattern, and processing pattern. The multivariate logistic regression model results showed that compared with the vegetarian pattern T2 group, the T1 group had lower risk of low birth weight (OR=0.56, 95%CI : 0.41-0.83), and the T 3 group had higher risk of low birth weight in offspring ( OR=2.32, 95%CI : 1.59-3.89); compared with the traditional pattern T2 group, the T3 group had higher risk of premature (OR=2.62, 95%CI :1.58-5.01); compared with the balanced pattern T2 group, the T3 group had a lower risk of spontaneous abortion (OR=0.73, 95%CI: 0.36-0.89); compared with the processing pattern T2 group, the T3 group had a higher risk of spontaneous abortion ( OR=1.97, 95%CI : 1.36-3.34) and higher risk of stillbirth ( OR=2.96, 95%CI : 1.49-6.26), and the T 1 group had a lower risk of stillbirths in offspring ( OR=0.52, 95%CI :0.33-0.83). Conclusion The women of childbearing age in Shaanxi have different dietary patterns, and there may be a correlation between dietary patterns and adverse pregnancy outcomes.

OBJECTIVE: To carry out Sanger sequencing for MMACHC gene variants among 65 Chinese pedigrees affected with combined methylmalonic aciduria and homocysteinemia, and summarize their genetic and clinical characteristics and prognosis. METHODS: Clinical characteristics of the 65 children identified with Methylmalonic acidemia and homocysteinemia at the Children's Hospital Affiliated to Zhengzhou University (Zhengzhou Children's Hospital) from April 2017 to April 2022 were selected as the study subjects. Potential variants of the MMACHC gene were detected by direct sequencing of the PCR products. RESULTS: The median age of the 65 children was 3 months (14 days to 17 years old). These included 28 cases (43.08%) from neonatal screening, 11 cases (16.92%) with a history of jaundice, and 9 cases (13.85%) with various degrees of anemia. The main clinical symptoms included development delay, slow growth, epilepsy, hydrocephalus, lethargy, feeding difficulty, regression or decline in motor ability, recurrent respiratory infections, anemia, jaundice, respiratory and heart failures, hydrocephalus, limb weakness, and hypertension. Blood and urine tandem mass spectrometry screening has revealed increase of methylmalonic acid, propionyl carnitine, propionyl carnitine/acetylcarnitine ratio, and propionyl carnitine/free carnitine ratio to various extents, and blood homocysteine was increased in all patients. The detection rate of genetic variants was 98.46% (128/130), and in total 22 types of MMACHC gene variants were detected. The most common ones have included c.609G>A (W203X) (58/128), c.658-660del (K220del) (19/128), and c.80A>G (Q27A) (16/128). Two novel variants have been identified, namely c.565C>T (p.R189C) and c.624_ 625delTG (p.A208Afs), which were respectively predicted as likely pathogenic (PM2_Supporting+PM3+PP2+PP3) and pathogenic (PVS1+PM2_Supporting+PM3+PP2) based on the guidelines from the American College of Medical Genetics and Genomics (ACMG). Exon 4 had the highest frequency for the detection. CONCLUSION Identification of MMACHC gene variants has confirmed the diagnosis in the children, among which the c.609G>A variant has the highest frequency. Discovery of the new variants has enriched the mutational spectrum of the MMACHC gene.
Humans ; Amino Acid Metabolism, Inborn Errors/genetics* ; Hydrocephalus ; Oxidoreductases