Objective The purpose of this study was to investigate the feasibility of short tandem repeat(STR) genotyping of cell free DNA in plasma for individual identification and paternity testing. Methods EDTA-Na2 DNA anti-coagulant blood samples were collected from 36 unrelated healthy volunteers,and both DNA in leukocytes and cell free DNA in plasma were extracted respectively using phenol-chloroform method. Target DNA in blood cells and plasma were amplified using regular STR typing and fluorescent multiplex STR assay separately,accordingly,the PCR products were analyzed by polyacrylamide gel electrophoresis and capillary electrophoresis. Results Using either normal PCR-STR or fluorescent multiplex STR assay,the consistent STR genotyping results were detected with similar efficiency for cell DNA and plasma DNA samples from the same individual. Conclusion Cell free DNA in plasma samples can be used as useful biological samples for STR genotyping,which can be applied to individual identification and paternity testing in forensic practice.

Objective To clone and express HCMV UL123 gene exon 2,3(ie1-exon2,3) in bacterial two-hybrid bait plasmid,and identify its self-activation property.Methods HCMV ie1-exon2,3 carried on pTWIN1/ie1 recombinant was amplified by PCR and cloned into the pBT plasmid,which was transformed into E.coli XL1-Blue MRF' Kan host strain.The positive recombinant was identified by PCR,restriction enzyme digestions and sequencing analysis.The verified plasmid was transformed into bacterial two-hybrid system reporter strain.The soluble fusion protein was analyzed by SDS-PAGE and Western blot.The self activation effect of the recombinant was then tested.Results Bacterial two-hybrid pBT/ie1-exon2,3 bait plasmid was successfully constructed.The corresponding soluble fusion protein rIE1-N_(85)/?C1 was expressed in bacterial two-hybrid system reporter strain,and didn't show self-activation property.Conclusion Bacterial two-hybrid pBT/ie1-exon2,3 bait plasmid without self-activation property was successfully constructed,and it can be used to screen the library of fetal brains.

Objective:To investigate the relationship between primary resection and KRAS gene mutation in the mild symptomatic patients with stage Ⅳ colorectal cancer,and to clarify its significance of prognosis.Methods:The clinical data of 46 mild symptomatic patients with stage Ⅳ colorectal cancer 2010 to December 2010 were collected.All the patients received primary resection.The KRAS gene mutation in the patients was detected by direct sequencing and the patients were followed up for 5 years.The influence of primary resection and KRAS gene mutation in prognosis of the patients with stage Ⅳ colorectal cancer was analyzed, and the clinical pathological features which might influence the prognosis were analyzed by survival analysis.Results:In 46 patients with colorectal cancer, KRAS gene mutation was found in 20 cases, the mutation rate was 43.4%, and most mutation was found at Codon 12. The KRAS mutation had relationship with the tumor site and multiple metastasis (P<0.05).The Kaplan-Meier survival curve and univariate analysis results showed that the median survival time of the patients with wild type of KRAS gene was 58.4 months, the median survival time of the patients with mutant type of KRAS gene was 42.2 months, and there was no significant difference (P>0.05).The median survival time of right colon cancer patients was 34.2 months, the median survival time of left colon cancer patients was 58.3 months, and there was sigificant difference (P<0.05).The cancer metastases including liver, lung and multiple metastasis were closely related to the poor prognosis of the colorectal cancer patients(P<0.05).The median survival time of the patients with stage Ⅳ colorectal cancer was 39.6 months after operation.Conclusion:After primary resection of the mild symptomatic patients with stage Ⅳ colorectal cancer,the median survival time of the patients with colorectal cancer in left colon site and right colon site were prolonged.Right colon cancer has more poorer prognosis than left colon cancer.KRAS gene mutation is associated with the tumor site and the multiple metastasis.The location of metastasis affect the prognosis.

Objective To investigate the clinical efficacy of the urethra URSL combined with micro-puncture and drainage treatment for pediatric calculus of lower urinary tract. Methods From January 2003 to January 2013, 66 cases with pediatric urinary tract calculi in our hospital were randomly equally divided into experimental group and control group. The experimental group was treated with transurethral bladder micro URSL combined puncture and drainage treatment, and the control group was given a simple transurethral URSL method of treatment. The operative time, postoperative urine turned clear time,pulling stone clearance after catheter time,postoperative hospital stay and the incidence of postoperative urethral stricture were observed and recorded. In addition, these data were compared between the two groups. Results The operations of two groups have successfully completed. The mean operative time, postoperative urine turned clear time, pull out the catheter stone clearance after time and postoperative hospital stay of control group was 30.2±2.45 minutes,2.5±0.5 days,2.1±0.8 days and 4.0±0. 5 days, respectively. Patients were followed up 1 year after operation, and 1 case of urethral stricture occurred. However, the mean operative time, postoperative urine turned clear time, pull out the catheter stone clearance after time and postoperative hospital stay of the test group was 20.36±2.35 minutes,1.5±0.7 days,1.1±0.25 days and 3.1±0.3 days,respectively,with no urethral stricture case occurred. There were significant differences between the two groups ( <0.05) . Conclusion Transurethral bladder URSL micro puncture and drainage combined with transurethral ureteroscopy lithotripsy treatment for children with lower urinary tract calculi has better efficacy and safety. It could shorter operative time, postoperative urine turned clear time, the gravel discharge time and average hospitalization time, reduce the incidence of postoperative secondary urethral stricture.

Objective To investigate the feasibility and safety of endoscopic treatment of ureteral calculi acute obstruction with urinary extravasation. Methods 56 patients with ureteral calculi acute obstruction and urinary extravasation were randomly divided into two groups:the treatment group and the control group,28 cases in each group. Patients in the treatment group were given URSL or percutaneous nephrostomy drainage, and the secondary fistula was given URSL stone clearance treatment. Patients in control group were given traditional ureterolithotomy treatment. The stone clearance rate, the average recovery time after surgery, postoperative wound infection rate and the abnormal rate of postoperative albumin were observed in two groups. Results In the treatment group,28 patients had no residual stones with mean postoperative recovery time of (5.2 1.3) days,postoperative fever was found in 3 cases,obvious abnormal postoperative albumin in 3 cases. In the control group,residual stones were found in 3 cases,the average recovery time after surgery was (7.9 2.6) days,postoperative fever was found in 10 cases, and obvious abnormal postoperative albumin in 11 cases. There were statistically significant differences in stone clearance rate, the average recovery time after surgery, postoperative wound infection rate and the abnormal rate of postoperative albumin between two groups (P<0.05) . Conclusion Endoscopic treatment of ureteral calculi acute obstruction and urinary extravasation has advantages including better efficacy, less trauma, less complications and quicker recovery.

OBJECTIVEExplore the hygienic status of indoor environment in railway air-conditioned passenger trains.

METHODSWe carried out a hygienic investigation of various air-conditioned passenger trains on total 10 railway lines in Shanghai, Chengtu, Kunming, Harbin, Urumchi and Guangzhou railway bureaus for two years between July, 1997 and Oct., 1999. The monitoring was conducted on difference train cars at different time, different railway status. The control groups were selected in the same condition.

RESULTSThe results of temperature and wind speed in air-conditioned trains both meet with the national standards GB 9673-1996 in summer and winter. The humidity in summer exceeded the national standards. Of different railways the status, the noise in four different cars except dining cars exceeded the national standards. The results of shock demonstrated that all measure up to the standards of ISO 2631/1-85. As to indoor air quality, the concentration of CO in air-conditioned trains in difference seasons(summer, autumn, winter) all met with the national standards, the ratio was 100%. In summer, the concentrations of CO2 in hard seat cars and soft berth car exceeded the national standard with a statistic significance(P < 0.01). In winter, the concentrations of CO2 in soft berth and hard berth cars both exceeded the same standard. The results of inhalable particulate matter(IP) and total count of bacteria all satisfied the standards, meanwhile the natural lighting and artificial illumination in air-conditioned train cars all were superior to that in ordinary trains.

CONCLUSIONHumidity in summer and noise in dining car exceeded the standards, the concentration of CO2 in different cars in summer and winter exceeded the standards in some degrees.

Air Conditioning ; Air Pollution, Indoor ; Carbon Dioxide ; analysis ; Environmental Monitoring ; Humidity ; Motor Vehicles ; standards ; Particulate Matter ; analysis ; Seasons

Atherosclerosis(AS), characterized with the accumulation of lipids on the vessel wall, is an immune-related inflammatory disease which promotes the progression of cardiovascular diseases(CVD). The imbalance of Treg/Th17 accelerates the progression of AS. Yangyin Huoxue Prescription(YHF)is an efficient traditional Chinese medicine used in the treatment of AS, but the effects of YHF on the balance of immunity have still not been clarified. This project was designed to investigate the effects of YHF on the imbalance of Treg/Th17 and AS in ApoE-/- mice induced by high-fat diet(HFD). ApoE-/- mice were given HFD to induce AS and administered low-dose YHF(18 g/kg)or high-dose YHF(36 g/kg)for 20 weeks. Atherosclerotic plaque area was analyzed by oil red O staining. Serum lipids were measured by biochemical kits. Treg or Th17 cells in peripheral blood were detected by flow cytometry. mRNA and protein expression of Foxp3 and RORγt of aortas were determined by qRT-PCR, Western blot and immunohistochemistry. Splenic CD4+T cells of mice were isolated and activated by anti-CD3/CD28, and then treated with lipopolysaccharide(LPS)and YHF. The expression of mRNA and protein of Foxp3 and RORγt were detected by qRT-PCR and immunofluorescence. It was found that YHF reduced the plaque area, decreased lipid level and increased the ratio of Treg cells in peripheral blood. Moreover, YHF increased mRNA or protein expression of Foxp3 in aortas in vivo or CD4+T cells in vitro while decreasing mRNA or protein expression of RORγt. These results suggested that YHF can regulate the imbalance of Treg/Th17 in ApoE-/- mice induced by HFD, and reduce the inflammatory stimulation of LPS on CD4+T cells, thereby improving AS.

Objective:To explore the efficacy of Matas test combined with endovascular intervention on patients with traumatic internal carotid-cavernous fistula.Methods:A retrospective case study was performed on 144 patients with traumatic internal carotid-cavernous fistula treated in our department from August 2012 to June 2018. The clinical symptoms, intraocular pressure and visual acuity data before and after surgery were counted, and the paired t test was used for statistical analysis. Logistic regression was performed to analyze factors affecting recurrence.Results:The main symptoms of traumatic internal carotid-cavernous fistula were proptosis, bulbar conjunctival hyperemia and cranial sound. Postoperative intraocular pressure was significantly lower than preoperative intraocular pressure [(11.13±2.97) mmHg vs. (22.37±6.64) mmHg] and postoperative visual acuity was significantly higher than preoperative visual acuity [(0.69±0.36) vs. (0.47±0.25)] and (both P<0.05). Postoperative intraocular pressure was an independent risk factor for recurrence within 3 months ( OR=0.357, 95% CI: 0.135-0.944, P=0.037), and the recurrence rate was 10.42%. Conclusions:Matas test combined with endovascular intervention in the treatment of traumatic internal carotid-cavernous fistula can effectively reduce intraocular pressure, improve visual acuity, and has a definite curative effect. It is one of the effective treatment methods for traumatic internal carotid-cavernous fistula.

Objective: To understand the nutritional literacy level and associated factors of primary and secondary school students in Beijing, so as to provide a scientific basis for improving student nutrition. Methods: From October 2022 to May 2023, a multi stage cluster random sampling method was employed to select a total of 14 568 primary, junior and senior high school students from 16 districts (ecluding the Economic Technological Development area) in Beijing. Through a survey questionnaire on nutritional literacy and dietary hehavior of school age children, basic information as well as data on nutritional literacy levels across four dimensions:nutrition related knowledge concepts, food selection, food preparation, and food intake dimensions were obtained. The Wilcoxon rank sum test, Kruskal-Wallis test, Spearman correlation analysis, Chi square test and binary Logistic regression were used for the analysis. Results: The median total score of nutritional literacy among primary and secondary school students in Beijing was 68.8. Approximately 26.0% of primary and secondary school students achieved nutritional literacy standards. The median scores and rates of meeting the standards for nutrition related knowledge concepts, food selection, food preparation and food intake dimensions were 23.0, 42.1%; 17.0, 27.4%; 6.5, 33.5%; 23.0, 33.3%, respectively. There were positive correlations between all pairs of the four dimensions ( r=0.33-0.49, P <0.05). The results of multiple Logistic regression analysis showed that primary school students, junior high school students, female students, suburban students, caregivers with a college education level and a bachelor s degree or above were the positive arrelation factors that promoted the achievement of nutritional literacy standards ( OR =2.21, 1.39, 1.18, 1.27, 1.42, 1.66, P <0.05). Conclusion The literacy level of primary and secondary school students in Beijing needs to be significantly improved. School stage, gender, region and caregiver s education level are associated factors.