OBJECTIVETo detect the methylenetetrahydrofolate reductase(MTHFR) gene C677T mutation with molecular beacon technique and assess the revant applicability.

METHODSA total of 228 samples were analyzed using molecular beacons which are oligonucleotide probes to become fluorescent upon hybridization. Wild-type molecular beacon and mutant beacon were designed to detect the genotypes of MTHFR gene.

RESULTSAnalysis of the 228 samples indicated that there were three genotypes including 41 homozygous mutants, 113 heterozygous individuals and 74 wild-type individuals. Every sample was identified clearly.

CONCLUSIONThe present method, a closed-tube PCR/hybridization assay, is a simple, high-throughput and fast procedure that is fully automated for detecting gene mutation.

DNA Mutational Analysis ; methods ; Fluorescent Dyes ; chemistry ; Genotype ; Humans ; Methylenetetrahydrofolate Reductase (NADPH2) ; genetics ; Oligonucleotide Probes ; chemistry ; genetics ; Point Mutation ; Sensitivity and Specificity

Objective To investigate the expression of Toll-like receptor(TLR) 9 of circulating plasmacytoid dendritic cell (pDC) and analyze the frequency and interferon (IFN)-?production of circulating pDC during hepatitis B virus(HBV) infection.Methods Peripheral blood was collected from 69 HBV-infected patients,including 14 cases of asymptomatie HBV infection,30 cases of chro- nic hepatitis B(CHB),25 cases of HBV-related liver cirrhosis,and 21 healthy blood donors as con- trols.Flow cytometry was used to analyze the frequency of circulating pDC and the mean fluorescence intensity(MFI) of TLR9.Fresh peripheral blood mononuclear cells(PBMC) were stimulated by CpG ODN 2216 for 24 h in vitro.IFN-?in the supernatant was measured using enzyme-linked immunosorbent assay(ELISA) to analyze the frequency and IFN-?production of pDC during HBV in- fection.Data were analyzed with SPSS 13.0 for windows.Results Compared with healthy controls (62.6?10.7),the MFI of TLR9 of patients with asymptomatic HBV infection,those of CHB and HBV-related cirrhosis were significantly reduced (P

In order to develop a rapid detection kit for novel avian influenza virus (AIV) subtype H7N9, two sets of specific primers and probes were designed based on the nucleotide sequences of hemagglutinin antigen (HA) and neuraminidase antigen (NA) of novel H7N9 virus (2013) available in GenBank to establish the method of TaqMan probe-based multiplex real-time RT-PCR for rapid detection of AIV subtype H7N9. The primer and probe of HA were for all H7 subtype AIVs, while the primer and probe of NA were only for novel N9 subtype AIVs. The results showed that this method had high sensitivity and specificity. This method was applicable to the testing of positive standard sample with a minimum concentration of 10 copies/microL; it not only distinguished H7 subtype from H1, H3, H5, H6, and H9 subtypes, but also distinguished novel N9 subtype from traditional N9 subtype. A total of 2700 samples from Zhuhai, China were tested by this method, and the results were as expected. For the advantages of sensitivity and specificity, the method holds promise for wide application.
Animals ; Birds ; virology ; Influenza A Virus, H7N9 Subtype ; genetics ; isolation & purification ; physiology ; Influenza in Birds ; prevention & control ; virology ; Real-Time Polymerase Chain Reaction ; methods ; Species Specificity ; Taq Polymerase ; metabolism ; Time Factors

Objetive: To investigate whether apoptosis of SGC7901 cells can be induced by the expression of the recombinant gene of anti-HER2 ScFv/tBid. Methods: The recombinant anti-HER2 ScFv/tBid gene was cloned into vector pCMV and the recombinant plasmid was transfected into SGC7901 cells. The gene expression was detected by RT-PCR and immunofluorescent staining. Cell counting was carried out to show the effect of the gene transfection on cell growth. At the same time, significant apoptotic peak was detected by flow cytometry in recombinant anti-HER2 ScFv/tBid gene transfected cells. Results: The fusion protein of anti-HER2 ScFv/tBid was observed in the cytoplasm of transfected SGC7901 cells. The transfected cells displayed typical cell growth inhibition and apoptosis. Conclusion: Fusion protein of anti-HER2 ScFv/tBid can induce apoptosis of SGC7901.

OBJECTIVETo modify the current PCR-based method for rapid and efficient preparation of small interfering RNA (siRNA) expression cassette to improve the efficiency of RNA interference.

METHODSThe U6 promoter sequence was amplified by PCR using the genomic DNA of K562 cells as the template, and cloned into pMD18-T vector which served as the template for further PCR amplification with the primers on the plasmid. The amplified product was directly used as the template for preparing siRNA expression cassette. The siRNA expression cassette targeting p53 gene was amplified, verified by sequencing, and transfected into SH-SY5Y cells. After a 48-hour transfection, the cells were harvested and the total RNA was for RT-PCR for evaluating the effect of RNA interference.

RESULTSThe sequencing result confirmed the correct U6 promoter sequence cloned from K562 cells. After transfection of SH-SY5Y cells for 48 h with siRNA expression cassette, the p53 gene expression was inhibited at the mRNA level in comparison with the control cells as demonstrated by RT-PCR detection.

CONCLUSIONThe siRNA expression cassette prepared using the established method described hereby can be well applicable in RNA interference research.

Gene Silencing ; Gene Targeting ; methods ; Genetic Vectors ; genetics ; Humans ; K562 Cells ; Polymerase Chain Reaction ; RNA, Small Interfering ; biosynthesis ; genetics ; RNA, Small Nuclear ; Tumor Suppressor Protein p53 ; biosynthesis ; genetics

OBJECTIVETo investigate the effect of lymphadenectomy adjacent to inferior mesenteric artery root on the prognosis of rectal cancer.

METHODSClinicopathological data of 260 cases with rectal cancer undergone radical operation were analyzed retrospectively. The patients were divided into two groups. Group D(2): the lymph nodes adjacent to mesenteric artery root were not excised (n=188). Group D(3): the lymph nodes adjacent to mesenteric artery root were excised (n=72). Prognosis of two groups was compared during the follow-up period.

RESULTSIn group D(2), the 1-, 3-, 5-year total survival rates (TS) were 97.3%, 87.2% and 77.1%, and tumor-free survival rates (TFS) were 93.1%, 83.0% and 76.8% respectively. In group D(3 ), the 1-, 3-, 5-year total survival rates (TS) were 94.4%, 79.2% and 73.6%, and tumor-free survival rates (TFS) were 86.1%, 76.4% and 71.0% respectively. The differences of TS and TFS between two groups were not significant according to Kaplan-Meier analysis (P>0.05). Multivariate analysis revealed that the excision of lymph nodes adjacent to mesenteric artery root was not statistically correlated with the recurrence, metastasis and survival time after radical operation of rectal cancer.

CONCLUSIONExcision of lymph nodes adjacent to inferior mesenteric artery root has no significant impact on prognosis and it is unnecessary in the radical operation of rectal cancer.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lymph Node Excision ; methods ; mortality ; Lymph Nodes ; surgery ; Lymphatic Metastasis ; Male ; Mesenteric Artery, Inferior ; surgery ; Middle Aged ; Prognosis ; Rectal Neoplasms ; mortality ; pathology ; surgery ; Survival Rate ; Treatment Outcome

The automatic cleaning machine we have developed, adopts a SCM system in automatic cleaning. The machine has five functions: ultrasonic cleaning, cold or hot water spraying, drying and greasing. The clinical applications show that the machine with a good effectiveness is suitable for the cleaning of many surgical instruments. It also raises working efficiency, cuts down on the cost of repair and maintenance and reduces the injury and infection to nurses caused by manual cleaning, satisfying the needs of clinical applications.
Automation ; instrumentation ; Disinfection ; instrumentation ; Equipment Design ; Surgical Instruments ; standards ; Ultrasonics ; instrumentation

OBJECTIVESTo investigate the effects of astilbin on the expressions of TNF alpha and IL-10 during liver warm ischemia-reperfusion injury.

METHODSC57BL/ 6 mice were randomly divided into 4 groups (n = 8): sham-operated group (Sham), model control group(I/R), low dosage of astilbin treatment group (10 mg/kg) and high dosage of astilbin (40 mg/kg) treatment group. The treatment group mice were intraperitoneally injected with 10 or 40 mg/kg astilbin 24 hours and one hour before Ischemia, the hepatic ischemia-reperfusion model were thus established. After jn90 of min ischemia and 6 h reperfusion of the partial hepatic lobe, the expressions of TNF alpha and IL-10 in liver tissues collected from the experimental groups were detected by Western blot and semiquantitative RT-PCR.

RESULTSThe expression of TNF alpha protein in liver tissues gradually decreased in treatment groups (low and high dosages of astilbin treatment groups) as compared to the I/R model control group. Similar results were observed in the mRNA expressions of these genes as determined by semiquantitative RT-PCR (P less than 0.05 for low dosage group; P less than 0.01 for high dosage group). Compared with the I/R model control group, the expression of IL-10 was increased in both treatment groups (low dosage group P less than 0.05; large dosage group P less than 0.01).

CONCLUSIONTreatment with astilbin decreases TNF alpha expression but induces IL-10 expression in liver during warm ischemia-reperfusion injury.

Animals ; Flavonols ; pharmacology ; Interleukin-10 ; metabolism ; Liver ; drug effects ; metabolism ; Male ; Mice ; Mice, Inbred C57BL ; Reperfusion Injury ; etiology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism ; Warm Ischemia

To improve the recognition of angioimmunoblastic T-cell lymphoma (AITL) and to reduce misdiagnosis, a case diagnosed as AITL with large granular lymphocytosis was reported and the related articles were reviewed. A series of clinical tests, pathologic examination and immunohistochemical test, TCR gene rearrangement detection by multiple PCR and assay of lymphocyte immunophenotypes were carried out. The results indicated that the patient was characterized by fever, skin rash, generalized lymphadenopathy, splenomegaly, pleural effusion, ascites, anemia and thrombocytopenia, increase of circulating large granular lymphocytes with CD3(-) and CD16(+), CD56(+) were detected, T-cell receptor γ-chain gene was rearranged. More large granular lymphocytes with abnormal mitosis were found in ascites. The histological and immunohistochemical changes observed by the lymph node biopsy were compatible with AITL, some cells of which were CD56-positive. In conclusion, AITL is characterized by aggressive progress and generally occurs in elderly patients, its clinical prognosis is poor, the large granular lymphocytosis may be an autoimmune response to the tumor cells or originate from tumor stem/progenitor cells.
Humans ; Immunoblastic Lymphadenopathy ; complications ; immunology ; pathology ; Immunophenotyping ; Leukemia, Large Granular Lymphocytic ; complications ; immunology ; pathology ; Male ; Middle Aged

OBJECTIVESTo study the knowledge, attitude and practice (KAP) concerning asthma in parents of children with asthma and the related factors.

METHODQuestionnaire survey was conducted by using the multi-center research methods in twenty hospitals with asthmatic clinic or pediatric clinic in Beijing from August to October in 2008. The investigation focused on Beijing permanent residents with children who have been diagnosed as asthma for more than 6 months.

RESULTTotally 390 parents of asthmatic children were investigated among whom 351 provided valid questionnaires, the response rate reached 90.0%; 75.2% of the parents knew that the nature of asthma is a chronic allergic airway inflammatory disease; 54.4% of parents think that even if the child is in a state of asthma control he/she can participate in only mild exercise; 88.4% of parents consider that the long-term use of inhaled corticosteroid would have a mild effect on growth and development of children. Of the asthmatic children, 32.5% adhered to use inhaled corticosteroid prescription; 47.2% adhered to use leukotriene receptor modulator; 45.3% of parents of children with acute asthma attack will add antibiotics; 18.1% of parents would choose short-acting beta2-agonist. The multi-factors Logistic regression suggested that the parents' educational background and the frequency of referral in children with asthma under control may have an influences on the parents' KAP (P < 0.05, OR > 1).

CONCLUSIONThe general understanding of asthma in asthmatic children's parents in Beijing area is good, the higher the parents' educational background and revisit frequency of asthmatic children whose disease is under control, the better the Parents' KAP.

Adolescent ; Asthma ; Child ; Child, Preschool ; China ; Health Knowledge, Attitudes, Practice ; Humans ; Infant ; Parents ; Surveys and Questionnaires