The etiological agent of severe acute respiratory syndrome (SARS) was identified as a new coronavirus, termed SARS-CoV. Establishment of an efficient and sensitive diagnostic system of SARS-CoV genetic materials is crucial for SARS control. In this study, we quantified SARS-CoV mRNAs in both infected cell culture lysate and in supernatant by using Real-time quantitative revere transcription-PCR based on EvaGreen鈩?dye and Taqman-MGB probes. For extensive evaluation of sensitivities and specificities, 13 pairs of primers and 4 probes were designed based on different genes of SARS-CoV. Glyceraldehydes-3-phosphate dehydrogenase (GAPDH) was selected as the internal control gene. Results showed that S-gene-specific PCR was the most sensitive for detection, but because of its sequence variability in the different viral strains, primers and a probe based on the N gene were suitable substitutions. Meanwhile, we found the mRNA concentrations in cell culture lysates were much higher than in cell supernatant and facilited more sensitive detection of the SARS-CoV.

Objective To explore the potential changes of connexin Cx36 in hippocampus and cortical neurons of rats with hyperthermia -induced convulsion.Methods Rats were divided into 2 groups according to the random number table method:normal control group and experimental group.Febrile convulsion model was elicited through im-mersion in warm water.The experimental group was generated following febrile convulsion model:hyperthermia group and febrile convulsion group.Among normal control group,hyperthermia group and febrile convulsion group,western blot analysis and immunofluorescence labeling techniques were used to examine the expression of Cx36 protein in the hippo-campus and cortex area.One -Way ANOVA was used to compare the mean of multiple sample,the LSD test was used to compare the two means.Tamhane′s test was used when variance were uneven.Results The incubation period,seizure duration and temperature were (4.39 ±0.08)min,(5.38 ±0.07)min,(41 .87 ±0.06)℃ after hyperthermia-in-duced convulsion,respectively.Western blot analysis showed that the expression of Cx36 protein in the hippocampus and cortex area decreased gradually after 1 0 times of seizure in normal control group,hyperthermia group and febrile convulsion group,and the febrile convulsion group decreased most obviously.Compared with normal control group and hyperthermia group,respectively,in febrile convulsion group Cx36 expression obviously decreased in the hippocampus and cortex in rats with 1 ,5,1 0 seizure times induced by hyperthermia,and with the increase of number of induced con-vulsion,the expression of Cx36 was significantly decreased in the cortex (0.1 04 ± 0.01 2)and CA1 (0.091 ± 0.01 1 ),CA3 (0.090 ±0.01 1 )and DG (0.092 ±0.01 2)areas of hippocampal neurons compared with the normal control group (0.21 2 ±0.01 7,0.1 67 ±0.01 3,0.1 59 ±0.01 4,0.1 71 ±0.01 3)and the hyperthermia group (0.1 89 ± 0.006,0.1 44 ±0.008,0.1 29 ±0.005,0.1 65 ±0.01 1 )(all P <0.05).Furthermore,the extent of reduction in Cx36 expression seemed to correlate with the number of seizures.Conclusion With the increase of thermal seizure frequen-cy,Cx36 expression of rats was decreased obviously which may lower convulsion threshold and lead to recurrent seizures.

Background and purpose:miR-101 has been reported to be down-regulated in gastric cancer, colorectal cancer, breast cancer as well as prostate cancer acting as a tumor suppressor gene. However, its function in ovarian cancer is still unknown. The aim of this study was to investigate whether miR-101 can suppress cell growth and invasion of ovarian cancer cells by targeting DNMT3A, so as to reveal molecular mechanism to inhibit ovarian cancer. Methods:Quantitative real-time palymerase chain reaction (qRT-PCR) method was employed to detect the expression of miR-101 in ovarian cancer and cancer adjacent normal ovarian tissues. SKOV3 cells were transfected with miR-101 mimics, and DNMT3A siRNA was transfected as a positive control. Then Western blot was used to detect the expres-sion of DNMT3A protein regulated by miR-101 in SKOV3 cells. The growth and invasion ability of SKOV3 cells were evaluated by MTT and Transwell invasion assays.Results:qRT-PCR showed that miR-101 was down-regulated in ovarian cancer tissues. Western blot showed that the level of DNMT3A protein was inhibited by restored miR-101 or knock-down of DNMT3A in SKOV3 cells. Following transfection of miR-101 mimics or knock-down of DNMT3A for 48, 72 and 96 h respectively, MTT assay showed that theD values were signiifcantly lower than the control group, (P<0.05). After transfection of miR-101 mimics or knock-down of DNMT3A for 36 h, Transwell invasion assay showed that the numbers of cells through the basement membrane was (105±7) and (107±13), respectively, which are signiifcantly different from the control group (213±11), indicating invasion of SKOV3 cells signiifcantly slowed down (P<0.05).Conclusion:miR-101 suppresses cell growth and invasion by targeting DNMT3A in ovarian cancer.

To investigate the relationship between the expression of early growth response gene 1 (EGR-1) and p38MAPK pathway in the paclitaxel resistance of ovarian carcinoma cells, the effect of p38MAPK inhibitor SB203580 on cell apoptosis was examined by using Hoechst 33258 staining. The intracellular Rh123 (Rhodamine 123) accumulation was detected by the flow cytometry (FCM). The 50% inhibition concentration (IC50) of paclitaxel for A2780/Taxol cells was determined by MTT method. Electrophoretic motility shift assay (EMSA) was employed to examine the EGR-1DNA binding activity. MDR1 and EGR-1 mRNA were assessed by RT-PCR. The expressed of p-gp, phosphorylated p53 and p38 were detected by Western blotting. SB203580 could remarkably promote the apoptosis of A2780/Taxol cells, and the cell apoptosis was in a time-dependent manner. Cellular Rh123 accumulation was increased, and the IC50 of paclitaxel for A2780/Taxol cells was decreased significantly. A2780/Taxol cell line after SB203580 treatment was shown to have a significantly higher level of EGR-1 DNA binding activity. SB203580 down-regulated the activity of p38MAPK pathway, but up-regulated EGR-1 expression. SB203580 significantly increased the level of cellular phosphorylated p53 protein, but decreased the p-gp protein level and MDR1 mRNA level in A2780/Taxol cells. There existed a close relationship between p38MAPK pathway and the paclitaxel resistance of ovarian carcinoma cells. The expression of EGR-1 mediated by p38MAPK pathway plays a critical role in paclitaxel resistance of ovarian carcinoma cells.

Objective To evaluate the efficacy and safety of diacerhein in the treatment of patients with knee osteoarthritis.Methods The efficacy and safety of diacerhein was randomly investigated in 42 pa- tients with knee osteoarthritis using parallel group methodology and a double-dummy technique to ensure dou- ble blind status with respect to diacerhein and control drugs diclofenac.Results Significant changes were ob- served in 20 meters walk pain,knee joint tenderness scale,WOMAC index scale,5F-36 health survey,knee joint swelling scale,compared with baseline(P＞0.05)in both diacerhein and diclofenac group respectively.No difference was found between diacerhein group and diclofenae group.The patient global assessment and physi- cian's global assessment were similar in diacerhein group and diclofenac group(P＞0.05).The side effect was similar in two groups.All of these side effects in gastrointestinal tract appeared to be transient.Conclusion Diacerhein can effectively relieve pain and swelling of knee osteoarthritis,and provides us a new effective and safe approach for treating knee osteoarthritis.

Objective To analysis the quality of life of patients with active ankylosing spondylitis (AS).Methods The quality of life was assessed in 52 patients with active AS using SF-36 and was compared with the general population.The correlation between the quality of life and the clinical measures of disease,in- cluding the Bath AS disease activity index(BASDAI),Bath AS functional index(BASFI),Bath AS metrology index(BASMI),patient's global assessment(PGA),spinal inflammation,total back pain,nocturnal back pain, and enthesis index(EI),were determined.Results The patients with active AS reported significant decreased scores on all dimensions of SF-36.The score of physical health components was poorer than that of mental health components.BASFI was the strongest factor correlated with the score of SF-36,followed by BASDAI and PGA.BASMI and total back pain were correlated with three dimensions only.In multivariate regression analysis,BASFI showed relative closer relationship to the qulity of life with active AS than other clinical mea- sures of disease and it accounted for 50.3%,35.2% and 47.9% of the variance in the physical health compo- nents,the mental health components and the overall score of SF-36,respectively.Conclusion The quality of life in patients with active AS is significantly declined compared with general population.The physical aspects seem to be more severely affected.Functional status of the patients with active AS are correlated with the quality of life closely.

Objective To analyze the relationship between the radiation doses to mediastinal lymph nodes regions and the regional failure patterns when involved field radiation therapy (IFRT) was used for limited-stage small cell lung cancer (SCLC).Methods The mediastinal lymph node regions (group 1 to 10) of the iimited-stage SCLC patients received definitive radiotherapy were contoured in treatment planning system.The intentional or incidental radiation doses to each lymph node regions were recorded.In-field recurrence,marginal recurrence and out-of-field recurrence were respectively defined as the volume of failed lymph nodes located within the 80％ iso-dose lines,in the 80％-20％ iso-dose lines and beyond the 20％ iso-dose lines of prescribed doses.Results A total of 1 216 lymph node regions in 76 patients were contoured.The median follow-up time was 17.4 months.At diagnosis,lymph node regions with metastatic rates ＞50％ were 4R (68.7％),4L (57.9％),10R (57.9％),2R (56.6％) and 7 (51.3％).The positive lymph node regions were all subjected to prescribed doses.The lymph node regions that received incidental radiation doses of more than 3 000 cGy were:3P,4L,7,6,4R,5,2L.The median lymph node failure-free time was 9.8 months.In this study,only 1 patient developed out-of-field mediastinal lymph nodes failure.The rest of out-of-field recurrences and marginal recurrences were developed in the supraclavicular regions or contralateral hila.Conclusions When IFRT is used to treat mediastinal lymph node regions for patients with SCLC,negative mediastinal regions can be subjected to considerable incidental radiation doses.Out-of-field recurrences of the mediastinal lymph node regions are rare.This is contributed by the incidental radiation dose to these regions.

Objective To explore the relationship between semen leukocyte and liquefaction and sperm motility through the anal‐ysis of routine semen examination results .Methods Retrospectively analyze on 8 666 cases of routine semen examination .Accord‐ing to the leukocyte count the semen samples which had sperms were divided into three groups ,Group A :white blood cells≤1 × 106/mL ,Group B:(1-4)× 106/mL ,Group C :>4 × 106/mL .Then compare the liquefaction time and sperm motility of these three groups .Results Among the 8 666 cases of semen analysis ,there were 164 cases of azoospermia(accounting for 1 .9% ) and 8 502 fine cases(accounting for 98 .1% ) .There were 7 419 ,1 014 and 69 cases in Group A ,B ,C respectively .In the three groups ,there were 5 323 ,740 and 50 cases of normal sperm motility respectively ;there were 2 096 ,274 and 19 cases of the abnormal motility ca‐ses respectively .In the three groups ,the normal cases of semen liquefaction time were 4 593 ,608 and 43 respectively ;the abnormal were 2 826 ,406 and 26 cases respectively .Statistical analysis showed no significant correlation between semen leukocyte and lique‐faction(P=0 .712) ,and between semen leukocyte and sperm motility(P=0 .486) .There were 1 217 cases of normal sperm motility and 4 027 cases of abnormal sperm motility in normal liquefaction group;there were 1 172 cases of normal sperm motility and 2 086 cases of sperm motility in abnormal liquefied group .There was statistically significant difference between the two groups(P=0 .000<0 .05) .Conclusion There were no correlation between semen leukocyte count and liquefaction or sperm motility ,but sperm mo‐tility and liquefaction are correlated .

Adolescent ; Cholesteatoma ; microbiology ; Ear, External ; Fistula ; etiology ; Humans ; Male ; Parotid Gland

Objective To investigate the relationship between folate metabolism-related gene polymorphism and fetal congenital defects,and discuss the effect of genetic factors on fetal congenital defects.Methods Retrospective analysis was used to investigate the genotype and gene frequency of 5,1O-methylenetetrahydrofolate reductase (MTHFR) C677T,A1298C gene loci and ethionine synthase reductase (MTRR) A66G gene locus in 132 cases of adverse pregnancy pregnant women (case group) and 150 cases normal pregnant women (control group) at the same period.The statistical differences were analyzed between the levels of their serum folate,vitamin B12 (Vit B12) and homocysteine (HCY).Results In the serum of case group,folate was positively correlated with Vit B12,and was negatively correlated with HCY,only HCY of skeletal system defects(6 cases) was higher (t =3.409,P ＜ 0.05).Comparing genotypes frequency of the MTHFR C677T,A1298C gene loci and MTRR A66G gene locus in case group with control group,the difference above was not statistically significant (P ＞ 0.05).In these three gene loci C/T,A/C and A/G allele frequency with the control group,the difference above was not statistically significant (all P ＞ 0.05).Different genotype combinations of MTHFR C667T and A1298C gene loci in control groups had no statistically different from the control group (P ＞ 0.05),and there was no synergy.Conclusions Maternal folate metabolism-related MTHFR and MTRR genes polymorphisms can affect the metabolic products levels accordingly.However,the correlation between the changes and the genetic mechanism of fetal congenital defects needs more large samples study in depth.