Objective To develop a gas chromatography-mass spectrometric method(GC/MS) for simultaneous determination of environmental estrogens including 4-nonylphenol,bishphenol A,diethylstilbestrol and 17?-ethinylestradiol and endogenic estrogens including 17?-estradiol,17?-estradiol,estriol and estrone in mutton. Methods Mutton samples were extracted using a mixture of acetonitrile-ethyl acetate(3∶2,V/V) and cleaned up by florisil cartridge. Derived with pentafluoropropionic anhydride(PFPA) ,the derivatives were determined by GC-MS. Results The linear range of determination was 0.2-5.0 ?g/kg. The intraday relative standard deviations of the peak area were 1.25%—5.85% and the detection limits ranged from 0.024 to 1.40 ?g/kg,and the rates of recovery ranged from 71.5%—111.0% except for 17?-ethinylestradiol( 55.4%) . Conclusion This method is simple,rapid,accurate and applicable to the determination of trace environmental estrogens in mutton

ObjectiveTo study the protective effects of 2,3,5,4'-tetrahydroxystilbene-2-O-β-D-glucoside(TSG) on the PC12 cells injury induced by H2O2.MethodsAll cells were divided into 5 groups: the saline control group(control group), Oxidative damage PC12 cells model group(H2O2 group) which was induced by H2O2, and TSG treated group, which oxidative damage PC12 cells model which was induced by H2O2 after given TSG 2 h at TSG 120 μg/L(TSG 120 μg/L+H2O2 group), 60 μg/L(TSG 60 μg/L+H2O2 group) and 30 μg/L(TSG 30 μg/L+H2O2 group) once. The survival rate of the cells was determined by MTT method, the content of lactate dehydrogenase (LDH) was determined by ultraviolet spectrophometry and the content of malonyldialdehyde (MDA) and superoxide dismutase (SOD) activity was measured respectively by thiobarbituric aicd and xanthine oxidese method. Immunohistochemitry method were used to detect the expression of bcl-2.ResultsTSG reduced obviously cells injury induced by H2O2. 30～120 μg/L TSG improved the cells survival rate, reduced LDH releasing and MDA content, increased SOD activity, and decreased the expression of bcl-2 in the PC12 cells injury induced by H2O2(P<0-05, P<0-01). ConclusionTSG has significantly potective effect on the PC12 cells injury induced by H2O2.

Sacral fractures are usually caused by high-energy violence,often complicated with injury to the lumbosacral plexus.In severe cases,they may cause instability of the posterior pelvic ring or of the lumbosacral junction.Their classifications commonly used in clinic are Denis,Tile,Isler and refined Denis type Ⅲ ones proposed by Roy-Camille and Gibbons.Recently,classifications of the lumbosacral lesions and scoring systems of the injury severity have often been used clinically.At present,internal instrumentation of sacral fractures is developing along the direction of high efficiency,safety and minimal invasion,but different methods of internal fixation have their own indications which should be strictly followed.This review summarizes the classifications of sacral fractures and their treatment advances.

OBJECTIVE To investigate the prevalence of erythromycin resistance genes ermB and mefA and the relationship of drug resistance and genes in Streptococcus pneumoniae.METHODS Forty three strains of S. pneumoniae were collected from respiratory system infected children from Dec 2004 to Oct 2005 at Yuying Pediatric Hospital of Wenzhou Medical College.Erythromycin sensitivity test was done by using MIC method.The erythromycin resistance genes ermB and mefA were detected by PCR.RESULTS In all forty three strains,forty were erythromycin resistant(93%),three were erythromycin sensitive.The total detection rate of erythromycin resistance genes ermB and mefA was 76.7% and 23.3%,respectively.There were neither gene ermB nor gene mefA in 3 erythromycin-sensitive S.pneumoniae.In 40 strains the detection rate of gene ermB was 82.5% and that of gene mefA was 25%.The erythromycin resistance gene ermB or mefA were detected in 35 of the 43 strains.The total detection rate of erythromycin resistance gene was 81.4%.In the 35 erythromycin resistance strains there were 25 strains in which gene ermB existed lonely and 2 strains in which gene mefA existed lonely.There were both genes ermB and gene mefA in 8 of the 35 erythromycin resistance strains.CONCLUSIONS The erythromycin resistance of S.pneumoniae can be caused mainly by gene expression of ermB or mefA,but the gene mefA seems to be less important than gene ermB.Obviously the erythromycin isn′t useful in treating S.pneumoniae infection.

Objective To evaluate the effectiveness of mindfulness-based stress reduction( MBSR) on anxiety and depression in spouses of breast cancer patients and to analyze the mediating effect of caregiver burden for the training effects. Methods Randomized control trial was used in this research.100 spouses of breast cancer patients were recruited and they were randomized into experimental group or control group. 91 of them completed the study finally (45 in experimental group and 46 in control group). Participants in con-trol group received no intervention,while participants in experimental group received MBSR once a week for 8 weeks. All participants were assessed with the Zarit Caregiver Burden Interview ( ZBI,measuring caregiver burden) ,Beck Depression Inventory( BDI,measuring depression level) and Hamilton Anxiety Scale( HAMA, measuring anxiety level) before and after intervention. Results After 8-week intervention,participants in experimental group got significantly lower total scores of ZBI,BDI and HAMA compared with pretest ((22.31 ±5.76) vs (25.98±6.00),(8.47±3.21) vs (11.87±3.51),(14.98±4.75) vs (17.93±4.65), P<0. 01), while there were no significant differences between posttest and pretest on any score in control group (P>0.05).Caregiver burden played mediating effect in MBSR for improving depression,and mediating effect was 56.6% of the total effect. Conclusion MBSR can effectively improve anxiety and depression with the spou-ses of breast cancer patients,and the caregiver burden is the intermediary variable in the effect of MBSR for improving depression.

Objective To retrospectively analyze the clinical characteristics ,pathological features and aetiology of pulmonary carcinoid tumourlets .Methods The clinical data of 10 patients with pulmonary carcinoid tumourlets in the West China Hospital of Sichuan University from April 1976 to October 2013 were collected for conducting the retrospective analysis and combining the lite‐raute review .Results Of the total 10 patients ,3 cases were male and 7 cases are female .9 cases were accompanied by bronchiecta‐sis ,1 case was complicated with lung adenocarcinoma .The clinical manifestations were lack of specificity ,which were dominated by repeated cough ,expectoration ,dyspnea and hemoptysis .The microscopic pathological examination suggested that there were differ‐ent degrees of inflammation reaction or fibrous tissue hyperplasia .All patients had no carcinoid syndrome ,lymph node and distant metastasis .10 cases received the surgical treatment .6 cases were pulmonary hypoplasia ,in which 1 case was carcinoid tumourlets complicating pulmonary sequestration .The postoperative follow up lasted for 5 - 80 months .8 cases had no recurrence and 2 cases were loss of follow up .Conclusion For the middle‐aged women patients with bronchiectasis and poor therapeutic effect ,complica‐ting carcinoid tumourlets should be vigilant ,active surgery combined with the pathologic examination is a suitable clinical decision .

Objective:The clinical manifestations, types of gene mutations, therapeutic effects and prognostic characteristics of 15 children with cblC type methylmalonic acidemia (MMA) and hydrocephalus were analyzed to improve the clinical understanding of the disease, so as to provide a basis for the treatment of the disease.Methods:From April 2015 to January 2019, 15 patients with MMA and hydrocephalus in Department of Pediatric Surgery, Peking University First Hospital were enrolled, and all gene detection showed clbC type.All the 15 patients underwent ventriculoperitoneal shunt.After surgery, the clinical manifestations and imaging findings were applied as the basis to adjust the pressure of the diverter valve appropriately.Clinical data and gene mutation characteristics of 15 children with cblC type and hydrocephalus were retrospectively analyzed, and the therapeutic effects and prognosis were summarized and analyzed as well.Results:There were 8 males (53.3%) and 7 females (46.7%), aged from 2 to 33 months.All the cases were followed up from 11 to 55 months, without death case and serious postoperative complications of hydroce-phalus.The head circumference of 3 cases (20.0%) was in the normal range, 1 case (6.7%) was greater than the normal range, and 11 cases (73.3%) were less than the normal range.Four patients (26.7%) were transferred to the pediatric intensive care unit after surgery.c.609G>A mutation was the most common in this study, with 7 cases (46.7%) of c. 609G>A homozygous mutation, and 5 cases (33.3%) of c. 609G>A heterozygous mutation.Clinical symptoms of intracranial hypertension were relieved or disappeared.The head circumference progressive enlargement was stopped.The anterior fontanelle tension greatly decreased, all " setting-sun" sign of eyes disappeared, and vision loss and hearing loss were better compared with the pre-operation.Four cases (26.7%) displayed normal intelligence and exercise, and 11 cases (73.3%) were left with mild to severe psychomotor retardation.During the follow-up pe-riod, the head CT showed that the ventricle was remarkably narrowed, and interstitial brain edema obviously improved.Conclusions:Ventriculoperitoneal shunt in the treatment of cblC type MMA with hydrocephalus has positive effects.The head circumference of most cblC type MMA with hydrocephalus is less than the normal range.c.609 G>A is the most common mutation in cblC type MMA with hydrocephalus.Perioperative " metabolic crisis" can result in serious complications.

Objective To study the relationship between the variation of the 20 exon of leptin receptor (LEPR) and lipid metabolism of the children with obesity. Methods Polymerase chain reaction-restricted fragments length polymorphism (PCR-RFLP) and polyacrylamide gel electrophoresis were used to analyze the variation of the 20 exon of the LEPR gene of the obesity group (120 obese children) and the control group (120 healthy children). Their serum triglyceride (TG), total cholesterol (TC), high density lipoprotein cholesterol (HDL-C), height, weight measured and the body mass index (BMI), fat percent (%fat) were calculated. Results 3 genotypes of the 20 exon of LEPR gene has been detected. Compared with the control group, the frequency of the gene variation at 3057 nucleotide G→A transversion is higher (P<0.05).The concentration of the serum TG, the level of BMI and %fat of the obese children with A/A genotype were higher than that with the G/G genotype (P<0.01),but the level of the serum HDL was lower (P<0.01).To the children with A/G genotype, only their TG level in serum are higher than that with the G/G genotype (P<0.05). Conclusion There is polymorphism at 20 exon LEPR gene in children with obesity, which may affect the lipid metabolism and the fat distribution. The obese children with A/A genotype should pay more attention to their diet to avoid some diseases followed obesity.

Objective To transfect the recombinant mIL-28A adenovirus vector into lung adenocarcinoma cell line LA795 and research its anticancer activity. Methods Transfected the constructed mouse IL-28(mIL-28) recombinant adenovirus vector into LA795 cell line, detected with PCR, immunocytal fluorescence, Tunel, Annexin V and MTT. Results Transfected with rAd-mlL-28A into the LA795 cells, mIL-28A gene expression products mRNA increased obviously, IL-28 expression was detected in cells obviously,apoptosis cell number increased, and the growth of LA795 cells transfected with rAd-mIL-28A were inhibited obviously. Conclusion The recombinant miL-28A adenovirus vector we have constructed, which expresses IL-28 when transfected to lung adenocarcinoma cell line LA795, inhibits growth of carcinoma cell to some extent, and may work by promoting the apoptosis of cancer cells.

Objective To express mouse IFN-λ2 stably and study its biological activity. Methods Full-length of mIFN-λ2 cDNA was obtained by using RT-PCR from cells of mouse spleen stimulated by ve-sicular stomatitis virus(VSV) and then subcloned to eukaryotic expressing vector PCAGG-EGFP. The recom-binant was transfected into CHO cells. VSV * GFP-B16 system was used to measure the antivirus activity. The constructed cell line MDBK-Mxp-Luc was used to study the character of Mx1 protein induced by the mIFN-λ2. Results The recombinant pMD18-T-mIFN-λ2 was digested by two kinds of enzyme, Sac I and Xho I, to produce the fragment was of 582 bp, and of which the sequence analysis of sequence shows it was entirely consistent with the nucleotide sequences reported in GenBank. PCAGG-EGFP-mIFN-λ2 eukaryotic expressing vector was constructed successfully and expressed stably in CHO cells, and the mRNA of mIFN-λ2 was verified expressing in CHO-PCAGG-EGFP-mIFN-λ2 cell line by RT-PCR. The antivirus activity of in the supernatant secreted by the CHO-PCAGG-EGFP-mIFN-λ2 cell line was 10~4 AU/ml. The mIFN-λ2 pro-tein can could induce the expression of the antivirus protein Mx1, and the expression of Mx1 protein induced by mIFN-λ2 enhanced with time going, 9 to 12 hours achieved the peak, 24 hours vanished. Conclusion Gene cloning of mIFN-λ2 was successful. The eukaryotic expressing vector of mIFN-λ2 was constructed suc-cessfully and expressed stably in CHO cells, and its product has obvious antivirus activity in vitro. And the antivirus activity of the product was closely correlated with inducing expression of antivirus protein Mx1.