Objective To investigate the expressions of OCT4 and P27kip1 and their correlations in renal hyaline cell carcinoma. Methods A total of 24 samples of renal hyaline cell carcinoma were detected by immunohistochemistry. The rate of cells with positive labelling of OCT4 and P27kip1 and their intra cellular distribution were observed in renal hyaline cell carcinoma. The expression levels of OCT4 and P27kip1 were compared between different gender, age (<60y and≥60y) and cancer cell metastasis groups. Results The rates of cells with positive OCT4 and P27kip1 expressions were 66.7%and 75% in renal hyaline cell carcinoma respectively. The ratio of cells with high, middle and low expression of OCT 4 were 33.3%, 20.5%and 12.5%in renal hyaline cell carcinoma tissues. OCT4 was mainly expressed in cytoplasm and nuclei with a few was expressed in the cell membrane. The ratio of cells with high, middle and low expression of P27kip1 were 12.5%, 25%and 37.5%in renal hyaline cell carcinoma tissues respectively. The positive staining of P27kip1 was in the cytoplasm and cell membrane, and a small number of nuclei were expressed. There were no significant differences in OCT4 and P27kip1 between different age and gender groups. There were significant differences in OCT4 and P27kip1 expressions between patients with metastasis and patients without metastasis (P < 0.05). A negative correlation was found between OCT4 and P27kip1 expres?sions in renal hyaline cell carcinoma (rs=-0.408, P<0.05). Conclusion The expression of OCT4 is negatively correlated with the expression of P27kip1. Inhibiting expressions of OCT4 or P27kip1, especially knocking down P27kip1 in cytoplasm can be used as a new diagnosis and treatment of renal cancer gene therapy.

Objective:To explore the relationship between loneliness and attachment in college students and the mediating effect of rumination.Methods:Totally 545 students [239 males,306 females;average age (20 ± 3) years] were assessed with the Russell UCLA Loneliness Scale (UCLA),Experiences in Close Relationships Inventory (ECR,including attachment avoidance and attachment anxiety) and Nolen-Hoeksema Ruminative Response Scale (RRS).The structural equation model was used to research the mediating effect of rumination on relationship between loneliness and attachment.Results:The scores of UCLA,ECR and RRS were positively correlated each other (r =0.30,0.20,0.44,Ps ＜ 0.01) in college students.The inspection of the mediating effect found fit indices for hypothetical model were acceptable (x2 =16.69,df=5,NFI =0.98,IFI =0.99,TLI =0.96,CFI =0.99,AGFI =0.96,RMSEA =0.07).Path analysis revealed that attachment avoidance and attachment anxiety predicted directly loneliness (β =0.10,0.06) and rumination (β =0.05,0.16),and predicted loneliness by rumination (β =0.18).Conclusion:The rumination may be associated with loneliness and individual attachment and play a partial mediating effect between loneliness and attachment in college students.

Objective To discuss the clinical significance of the changes of endothelin-1 and troponin-1 levels in kawasaki disease after acute stage. Methods 60 patients of kawasaki disease in which 12 cases of patient were complicated with coronary artery disease and 48 cases of patient who were not complicated with coronary artery disease were enrolled this study. Blood plasma endothelin-1 and serum cardiac troponin I were determined in kawasaki disease acute phase and recovery phase and 30 healthy children. Results Acute phase ET-1 and cTnI [(90.19±3.43)ng/L, (1.35±0.14)μg/L] and recovery phase ET-1 and cTnI [(89.09±2.44)ng/L, (1.12±0.11)μg/L ] in coronary artery lesions of (CAL) group had no significant difference（P＞0.05）. The concentration values of ET-1, cTnI in no coronary artery lesions (NCAL) group of acute phase (64.49±4.78)ng/L,(0.62±0.02)μg/L were different with convalescent phase (50.47±4.49)ng/L,(0.07±0.05)μg/L. There were significant difference between the the two phases (P＜0.01). Plasma endothelin-1 and serum troponin I concentrations were positively correlated in acute phase of Kawasaki disease in children (r＝0.93,0.96,P＜0.01). Conclusions Plasma endothelin-1 and serum cardiac troponin I test for the diagnosis of Kawasaki disease with coronary artery disease may be early indicators of risk monitoring, they are valuable indexes in diagnosis and treatment of Kawasaki disease.

Objective To study the effect of propofol intravenous anesthesia on T helper cells of patients with primary liver cancer during perioperative period.Methods A total of 86 patients with primary liver cancer in our hospital from November 2014 to October 2015 were selected,who were divided into observation group and control group according to the method of random numbers,43 cases in each group.The observation group were taken propofol intravenous anesthesia,and the control group were treated with sevoflurane inhalation anesthesia.The Th1 cells percentage,Th2 cells percentage and the ratio changes of the Th1/Th2 cells of two groups before anesthesia and postoperative 1 day were compared.The plasma cortisol levels of two groups before anesthesia,after anesthesia,intraoperative and 1 day after operation were observed.Results The percentage of Th2 cells in observation group and control group 1 day after surgery had no significant difference (P ＞ 0.05).The percentage of Th1 cells and Th1/Th2 cells ratio of the observation group were higher than those of the control group [(16.32 ± 1.76) ％ vs.(14.16 ± 1.03),(8.48 ± 0.92) vs.(7.11 ± 0.72)],the differences were significant (P ＜ 0.05).The plasma cortisol levels of observation group during operation and 1 day after operation were lower than those of the control group[(12.34 ± 1.02) μg/dL vs.(16.13 ± 1.26) μg/dL,(12.01 ± 0.94) μg/dL vs.(15.25 ± 1.08) μg/dL],the differences were significant(P ＜ 0.05).Conclusion The propofol intravenous anesthesia can encourage more Th to differentiate into Th1 cells,which plays a protective role in the patient's immune function.

AIM:To study the expressions of galectin-3 and the possible anti-infectious function in genital tract of female mouse. METHODS:The expressions of galectin-3 mRNA in different reproductive tissues were analyzed by RT-PCR. The mouse model of genital tract infection was constructed by injection of LPS and Freunds adjuvant incomplete into the vaginal mucous. The expression pattern of galectin-3 protein was detected by immunohistochemistry. RESULTS:The expression of galectin-3 mRNA was detected in the uterus,vagina,oviduct and ovary of normal adult mouse. The galectin-3 protein was expressed in the genital tract mucous of epithelial cell surface and uterine gland. After stimulation with LPS,the expression of galectin-3 protein was significantly up-regulated compared to normal control and adjuvant control (P

We analyzed the pathogenic characteristics of Salmonella strains isolated from diarrhea patients in Wuxi City,Jiangsu Province,China and compared the differences among pulse field gel electrophoresis (PFGE) patterns of main serotype strains,so as to provid scientific basis for disease control.After biochemical identification of the Salmonella strains isolated from infectious diarrhea patients in Wuxi in 2015,drug susceptibility test,serotyping and PFGE were applied to analyze these strains.Results showed that a total of 32 Salmonella strains were detected from 756 diarrhea specimens with a positive rate of 4.23 ％.The infection occurred more frequently between May and October and adults aged more than 60 years old affected mostly.There was no significant difference between genders in infected population.The drug susceptibility test indicated that the antibiotic resistance rate of these Salmonella strains to ampicillin (56.25 ％) was the highest,and to ciprofloxacin(6.25 ％)and Ceftazidime (6.25％) were the lowest.The 32 Salmonella strains belonged to 11 serotypes,and S.enteritidis(31.25％)and S.typhimurium(21.88％) were the predominant serotypes.PFGE showed that the pattern similarity of all S.enteritidis was more than 85 ％;PFGE patterns of S.typhimurium were different.In conclusion,the infection of Salmonella from diarrhea patients in Wuxi City had obvious season and age specific distribution,and the most prevalent serotype of Salmonella was the S.enteritidis.It is necessary to strengthen the surveillance of Salmonella concurrently in food and environment.

Objective To investigate the effects of TLR7 on imiquimod induced apoptosis of THP-1 derived macrophages.Methods Three cell lines ( THP-1 derived macrophages, MDCK cell line and HUVEC cell line) with different capabilities of expressing TLR7 were selected.The survival rates of cells af-ter the treatment with different concentrations of imiquimod were detected by MTT assay.The levels of IL-6 in the supernatants of TLR7 inhibitor chloroquine or TLR7-siRNA treated cells were detected by enzyme-linked immunosorbent assay.The apoptosis of cells was detected by flow cytometry after inhibiting the ex-pression of TLR7.Results Imiquimod induced the apoptosis of THP-1 derived macrophages, MDCK cell lines and HUVEC cell lines.The levels of IL-6 were significantly decreased as the expression of TLR7 was inhibited by treating THP-1 derived macrophages with chloroquine or TLR7-siRNA.Treating THP-1 derived macrophages with chloroquine or TLR7-siRNA did not affect the cell apoptosis induced by imiquimod.Con-clusion Imiquimod could induce the apoptosis of THP-1 derived macrophages through TLR7 independent pathway.

Objective To study the effects of Toll-like receptor 4(TLR4) on oxidized low density lipoprotein ( ox-LDL) induced macrophage apoptosis and its possible mechanism .Methods THP-1 derived macrophages were divided into four groups including untreated control group , ox-LDL treated group , ox-LDL+LPS treated group and tunicamycin treated group .MTT assay and flow cytometry analysis were performed to measure cell vitality and cell apoptosis , respectively .Oil red O staining was used to observe the phagocytosis of lipids by macrophages .The persistent and intense endoplasmic reticulum ( ER) stress markers were de-tected by analyzing the expression of glucose-regulated protein 78 ( GRP78 ) and CCAAT/enhancer-binding protein homologous protein ( CHOP) at mRNA and protein levels by q-RT-PCR and Western blot .Small in-terfering RNA ( siRNA) was used to silence the expression of TLR 4 to further elucidate its possible mecha-nism.Results Flow cytomotry and MTT assay showed that the number of apoptotic cells in ox-LDL+LPS treated group were increased more significantly than that in ox-LDL treated group (P<0.01), and cell apop-tosis in both two groups were greater than that in control group (P<0.01).Compared with control group, the expression of GRP78 and CHOP at mRNA and protein levels were up-regulated in ox-LDL+LPS treated group and ox-LDL treated group (P<0.01), and the expression of GRP78 and CHOP in ox-LDL+LPS treated group was significantly higher than that in ox-LDL treated group (P<0.01).Silenced expression of TLR4 al-leviated the endoplasmic reticulum stress (P<0.05).Conclusion Increased expression of CHOP contribu-ted to cell apoptosis .TLR4 might promote ox-LDL induced macrophage apoptosis through accelerating endo-plasmic reticulum stress .

A simple, rapid and sensitive colorimetric reverse-transcription loop-mediated isothermal amplification (RT-LAMP) was developed for rapid detection of Middle East respiratory syndrome coronavirus (MERS-CoV). The method employed six primers that recognized sequences of a nucleocapsid gene for amplification of nucleic acids under isothermal conditions at 63 degrees C for 60 min. Products were detected through a LA-320c Loopamp Turbidimeter (real-time RT-LAMP) or visual inspection of color change by pre-addition of Hydroxynaphthol Blue dye (visual RT-LAMP). Specificity of RT-LAMP was validated by detection of several human coronaviruses and common respiratory viruses. MERS-CoV real-time RT-LAMP had a linear correlation (R2) of 0.995 at 10(3)-10(6) copies. The limit of detection of real-time RT-LAMP, visual RT-LAMP and quantitative real-time PCR was 500, 1000 and 100 copies/reaction, respectively. The established RT-LAMP assay was demonstrated to be a rapid screening tool for MERS-CoV infection, and could be suitable in resource-limited clinical sites and for field studies.
Coronavirus Infections ; virology ; DNA Primers ; genetics ; Humans ; Middle East Respiratory Syndrome Coronavirus ; classification ; genetics ; isolation & purification ; Nucleic Acid Amplification Techniques ; methods ; Reverse Transcription

Objective To evaluate the role of opioid receptors in fentanyl-induced inhibition of proliferation and migration of human gastric cancer cell line MGC-803.Methods The human gastric cancer cell line MGC-803 was cultured in DMEM liquid culture medium.The cells were seeded in 6-well or 96-well plates and then randomly divided into 4 groups (n =54 each):control group (group C),fentanyl group (group F),naloxon group (group N) and naloxon + fentanyl group (group NF).The cells were exposed to 0.1 μmol/L fentanyl and 10 μmol/L naloxon in F and N groups,respectively.The cells were incubated with 10 μmnol/L naloxon for 30 min and then O.1 μmol/L fentanyl was added to the culture medium in group NF.The viability of the cells was detected by MTT assay after being incubated with fentanyl for 12,24,36,48,60 and 72 h.The cell apoptosis was assessed by flow cytometry after being incubated with fentanyl for 24 h.The migration of the cells was detected by wound healing assay after being incubated with fentanyl for 48 h.The proliferation of the cells was determined by colony formation assay at 7 day of incubation with fentanyl.Results Compared with group C,no significant changes in the viability of the cells,rate of colony formation,apoptotic rate and rate of cell wound healing were found in group N (P ＞ 0.05),and the viability of the cells,rate of colony formation and rate of cell wound healing were significantly decreased,and the apoptotic rate was increased in F and NF groups (P ＜ 0.05).There was no significant difference in the viability of the cells,rate of colony formation,rate of cell wound healing and apoptotic rate between group NF and group F (P ＞ 0.05).Conclusion Opioid receptors are not involved in fentanyl-induced inhibition of proliferation and migration of human gastric cancer cell line MGC-803 in vitro.