Objective To establish a method for detecting K-rag gene point mutation in pancreatic adenocarcinoma based on the pyrosequencing and to compare its performance with that of Sanger sequencing.Methods Genomic DNA was extracted from formalin-fixed,paraffin-embedded pancreatic tissues including 49 pancreatic adenocarcinoma,10 normal pancreas,11 chronic pancreatitis,18 benign pancreatic tumor,7 insulin carcinoma,9 ampullary carcinoma,7 bile duct carcinoma and 7 duodenum papillary adenocarcinoma tissues.K-rag gene point mutations at codon 12 were analyzed by pyrosequencing and Sanger sequencing,respectively.Results No mutant K-ras gene Wag detected in normal pancreas,chronic pancreatitis,benign pancreatic tumor,insulin carcinoma,ampullary carcinoma,bile duct carcinoma and duodenum papillary adenocarcinoma tissues by either pyrosequencing or Sanger sequencing.K-rag gene point mutation was detection rate in pancreatic adenocarcinoma tissues was 71.4%(35/49)by pyrosequencing and 61.2%(30/49)by Sanger sequencing,respectively.Conclusions Pyrosequencing is more sensitive than Sanger sequencing and is also accurate,rapid and of high throushput in detecting mutant K-ras gene.It may serve as a practical method for fast batch analysis of clinical samples.

Objective:This study aims to analyze the relationship between the effect of induction chemotherapy and the timing of radiotherapy in limited-disease or limited-stage small-cell lung cancer (LSCLC). Methods: Data from 148 LSCLC patients who re-ceived induction chemotherapy and radiotherapy between January 2009 and December 2012 were retrospectively analyzed. The effect of two to three cycles of induction chemotherapy was evaluated according to the RECIST version 1.1, which includes complete re-sponse (CR), partial response (PR), stable disease, and progressive disease. CR and PR were used to calculate response rate. The pa-tients were divided into early and late groups based on immediate radiotherapy after two to three cycles of induction chemotherapy. The survival rate was analyzed using the Kaplan-Meier method. Log-rank test and Cox regression model were used to evaluate the influenc-ing factors of the survival rate. Results: The median overall survival (OS) and progression-free survival (PFS) were 22.8 and 13.0 months, respectively. The early and late radiotherapy groups exhibited OS of 34.0 and 18.0 months, respectively, and corresponding PFS of 16.8 and 10.9 months. In the subgroup analysis, for the patients who responded to the induction chemotherapy, the early and late radiotherapy groups showed median OS of 18.0 and 19.5 months, respectively, and corresponding PFS of 19.4 and 11.7 months. For the patients who had no response to the induction chemotherapy, the early and late radiotherapy groups exhibited median OS of 18.0 and 9.5 months, respectively, and corresponding PFS of 12.4 and 10.3 months. Conclusion:All LSCLC patients who received two to three cycles of induction chemotherapy should receive radiotherapy as soon as possible after chemotherapy, regardless of their response to the induction chemotherapy.

Objective To evaluate the value of transvaginal ultrasound in diagnosing intrauterine adhesions.Methods Transvaginal ultrasound was performed in 136 patients with suspicious intrauterine adhesions and compared with hysteroscopy correspondingly .The ultrasonographic features of intrauterine adhesions on transvaginal ultrasound were summarized .Results One hundred and twenty one cases (89.0%, 121/136 ) of intrauterine adhesions were verified by hysteroscopy .The hysteroscopic findings included:(1) Forty seven cases(38.9%,47/121) were minimal intrauterine adhesions , 46 cases(38.0%, 46/121) were moderate intrauterine adhesions , and 28 cases (23.1%,28/121) were severe intrauterine adhesions.(2) Sixty one cases(50.4%,61/121) were central intrauterine adhesions , 24 cases(19.8%, 24/121) were marginal intrauterine adhesions , and 36 cases (29.8%, 36/121) were mixed type of intrauterine adhesions.The transvaginal ultrasound findings included:(1)Nineteen cases(40.4%,19/47) were minimal intrauterine adhesions ,33 cases(71.7%,33/46)were moderate intrauterine adhesions ,and 23 cases(82.1%,23/28) were severe intrauterine adhesions .(2) Thirty nine cases (63.9%,39/61) were central intrauterine adhesions ,9 cases(37.5%,9/24) were marginal intrauterine adhesions ,and 27 cases (75.0%, 27/36 ) were mixed type of intrauterine adhesions .By transvaginal ultrasound, seventy-five (62.0%,75/121) cases of intrauterine adhesions were correctly diagnosed , whereas 46 cases (38.0%, 46/121) were missed.And 3 cases ( 3.8%, 3/78 ) were misdiagnosed as intrauterine adhesions on transvaginal ultrasound,including one endometrial polyp ,one thin endometrium and one septate uterus .The sensitivity, specificity and accuracy of transvaginal ultrasound in diagnosing intrauterine adhesions were 62.0%(75/121), 80.0%(12/15) and 64.0%(87/136) respectively.There were significant statistical differences in diagnosing different degrees of intrauterine adhesions ( χ2 =15.956,P=0.000) and different parts of intrauterine adhesions( χ2 =8.792,P=0.012) by transvaginal ultrasound.Conclusions Transvaginal ultrasound is an effective, easy to perform and noninvasive technique in screening and diagnosing intrauterine adhesions.Transvaginal ultrasound is an effective way in diagnosing intrauterine adhesions showing a noninvasive and simpler way than hysteroscopy .Transvaginal ultrasound is of great value in screening and diagnosing intrauterine adhesions .

Objective To summarize and analyze prenatal ultrasound and postnatal autopsy ifndings in fetuses with urorectal septum malformation sequence (URSMS). Methods An analysis of prenatal ultrsound ifndings and postnatal autopsy features was performed on eleven cases of fetuses with URSMS that were identiifed by ultrasonography at Shenzhen Maternity&Child Healthcare Hospital in the period of January 2003 to December 2012. Results Prenatal ultrasonography showed a large abdominal cystic mass concomitant with imperforate anus in eleven fetuses with URSMS. The cyst contained unilocular or bilocular cystic structures in two fetuses, and trilocular cystic structures in nine fetuses. The cyst was demonstrated as clear acoustic transmission in three fetuses and unclear in eight fetuses. Out of them, seven fetuses had kidney abnormalities, six had ascites, and three had enterolithiasis. The associated systemic abnormalities included tethered cord in two fetuses, single umbilical artery in two fetuses, sacrococcygeal dysplasia in one fetus, and myocardial noncompaction in one fetus. 21-trisomy was found in one fetus by chromosome examination. Eleven cases were all identiifed as female fetuses by autopsy ifndings, including a single perineal opening and ambiguous genitalia with clitoral hypertrophy and labial fusion. The internal genital abnormalities included double vagina or longitudinal vaginal septum in nine fetuses, double uterus or uterus bicornis in ten fetuses and vaginal dysplasia in one fetus. Conclusions URSMS is a complex congenital malformation, which includes abnormalities of the urinary system, reproductive system and gastrointestinal track. An abdominal cystic mass visualized by prenatal ultrasonography might be the distinctive lesion in female with URSMS, and have an important diagnostic value. The kidney abnormalities and ambiguous genitalia can contribute to the diagnosis of URSMS.

The chemical structures of P1 A was identified by complete acid hydrolysis, partial acid hydrolysis, periodate oxidation-Smith degradation, methylation analysis, IR and NMR. The results showed that P1 A had a backbone consisting rhamnose, mannose, glucose and galactose. The side chain possessed arabinose and xylose. 1-->, 1-->6 and non-reducing terminal linkages existed in polysaccharide P1A, but there are doubling amount of 1-->2 and 1-->4 linkages. Oxidable linkage of P1 A accounted for 45%, and inoxidable linkage of P1A accounted for 55%. Mannose, glucose and galactose were mainly linked by 1-->2 linkage. Rhamnose, arabinose and xylose were mainly linked by 1-->2 and 1-->4 linkages. PlA contained beta-Glc(1,6)-,beta-Gal(1,3)-,beta-Man(1,4)-beta-Rha,-Glc(1,4)-, Glc(1)-,-Gal(1,4)- and Man(1)-.
Acanthaceae ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Molecular Weight ; Polysaccharides ; chemistry

Objective To study the awareness of Brucellosis prevention knowledge and prevalence of risk behaviors among the primary and secondary school students from breeding livestock’s families in Western pastoral areas of Jilin province,and to provide evidence for the prevention and control of Brucellosis.Methods With application of multi-stage random sampling method,a questionnaire survey which including Brucellosis prevention knowledge and risk behaviors was conducted among 209 primary and secondary school students in west part of Jilin province in 2012.Results 105(50.97%)students had heard Brucellosis among 206 students,the older students knew more about Brucellosis than the younger(χ2 =28.80,P=0.00).The M(Q)of the score of the Brucellosis-related knowledge was 8(6.00,9.00),the total passing rate was 28.64%.72(68.61%)students learned Brucellosis from village doctors and acquaintances,only 11(10.51%)students learned it from school;the contact rates of eight kinds of high-risk behaviors were helping feeding 21.84%(62/206),holding lamb 16.50%(58/206),cleaning sheepfold 8.73%(26/206),vaccinating sheep 5.34%(17/206),milking 2.91%(6/206);cleaning apoblema 2.91%(6/206),slaughtering 2.42%(5/206)and delivering lamb 1.94%(4/206);there was no significant difference between the passing rate and the eight kinds of the basic protective behaviors except feeding (P>0.05).Conclusion The level of Brucellosis prevention knowledge is low in Western pastoral areas of Jilin province, and there was a high-risk of contact behavior with sheep and low basic protection rate. It indicates that the education about prevention and control of Brucellosis in the students from breeding livestork’s familes should be strengthened in order to prevent Brucellosis infection.

Objective To investigate transcriptional expression and promoter CpG methylation status of A-kinase anchoring protein 12 (AKAP12) gene and analyze their correlation with clinical pathological stage in bladder transitional cell carcinoma. Methods AKAP12 mRNA expression level and promoter CpG metbylation status was measured by fluorescent quantitative RT-PCR (FQ-RT-PCR) and methylation specific PCR (MSP) in 30 bladder transitional cell carcinoma and adjacent normal tissues. The products of PCR were cloned and bisulfite sequenced. Results Decreased AKAP12 mRNA expression was demonstrated in 22 carcinomas (73. 3% ) and was significantly associated with turnout grade (P =0. 02).The frequency of promoter methylation of AKAP12 gene was 53. 3 % (16/30) and correlated with the tumor stage(r =0.52,Pn =0.03)and grade(r =0.61,Pn =0.01). Conclusion Aberrant promoter methylation of AKAP12 can result in the loss of gene expression and may association with bladder transitional cell carcinoma.

Objective To detect quantitatively AKAP12 methylation and evaluate its clinical significance in peripheral blood in colorectal cancer. Methods MS-HRM technology was used to detect quantitatively AKAP12 methylation in peripheral blood from 80 colorectal cancer patients and 20 healthy volunteers. They also validated the reproducibility and compared with MSP. Results Thirty-eight of the 80 colorectal cancer samples (47. 5% ) were found to be methylated at the AKAP12 promoter region by MS-HRM (the methylation levels of 24 cancer samples ranged between 1 % and 20% , the methylation levels of 12 cancer samples ranged between 20% and 60% , the methylation levels of 2 cancer samples ranged between 60% and 100% ). The methylation levels of 2 health samples were less than 10% . They also compared the results generated by MS-HRM with a traditional MSP assay. The AKAP12 MS-HRM assay was able to reproducibly detect 1% AKAP12 methylated DNA, whereas the MSP method was unable to detect less than 10% methylation. No significant correlation was observed between the AKAP12 methylation levels and patients' age and gender. However, AKAP12 methylation was significantly higher in DNA from colorectal cancer patients with high Dukes stage and differentiation (x2 =5. 93 or 8. 41, P = 0.01). Conclusions The authors demonstrate here for the first time, the utility of quantitative AKAP12 MS-HRM analysis of promoter methylation in peripheral blood samples. AKAP12 MS-HRM quantitative methods have many promising applications in the detection of colorectal cancer.

yggG, a Era-binding protein gene, was isolated and cloned from the E.coli genomic DNA library. Previous studies indicated that the product of yggG gene, YggG294(amino acids 1-294), strongly inhibited the growth of host bacteria and caused the death of bacteria cells. To elucidate whether Era is related to the death of bacterial cells expressed YggG294,A double promoter expression vector that can express YggG294 and Era proteins controllably in cells was constructed. Using this vector to express YggG294 and Era protein in the same E.coli cells, then analyzed the relation between YggG294 and Era. The results showed that the ratio of Era proteins to total proteins increased with the increase of induction time in E.coli cells without YggG294 expression and with little YggG294 expression;the ratio of Era proteins to total proteins seemed to be a constant level in E.coli cells overexpressing YggG294;but we could not detect any Era hydrolyzate in E.coli cells overexpressed YggG294 could not be detected. The results also showed that pre-expression of Era protein did not produce any effect on the growth inhibition of E.coli cells caused by YggG294. These results indicate that YggG294 can not hydrolyze Era protein in E.coli cells, and that YggG-Era interaction is not associated with the death of bacteria expressed YggG294. It is thus reasonable to draw a conclusion that Era is not associated with the growth inhibition of E.coli cells caused by YggG294. YggG294 inhibits the growth of bacteria by other way.

Carcinoma, Hepatocellular ; therapy ; Embolization, Therapeutic ; Humans ; Liver Neoplasms ; therapy ; Portal Vein