Objective To establish a rapid detection method for zika virus based on direct amplification re-al-time fluorescent quantitative reverse transcription polymerase chain reaction(RT-PCR)technique.Methods A direct amplification RT-PCR technique for the rapid detection of zika virus in 5 samples(whole blood,serum,saliva,throat swab and urine)was established by using a special function DNA polymerase and a preferred PCR enhancer.Results The detection limits of the 5 samples were 103 PFU/mL in serum,102 PFU/mL in urine,throat swab,and saliva,and 104 PFU/mL in whole blood.The coefficient of goodness-fit of stand-ard curves was above 0.98,and the amplification efficiency was 90％-110％.Zika virus nucleic acid was suc-cessfully amplified,but non-zika virus nucleic acid was not amplified.Based on the repeatable detection of sam-ples from urine,whole blood,and saliva,the variation coefficient of 6 repeated Ct values at 106 PFU/mL and 102 PFU/mL concentrations were all＜5％.The zika virus detection method established by the direct amplifi-cation RT-PCR technique was consistent with the detection results of conventional RT-PCR technique.Only two serum samples were detected in eight zika virus samples,and the remaining 62 non-zika virus samples and 12 negative samples were not amplified.Conclusion A rapid detection method for zika virus based on direct ampli-fication RT-PCR technique is successfully established.The method is simple,rapid,sensitive and specific.

Objective:To explore the effects of coenzyme Q10(CoQ10) on high-fat diet-induced obesity, lipid disorders, and bile acid metabolism in mice.Methods:Eight-week-old C57BL/6J mice were randomly divided into control group(regular chow), high-fat diet group(45% high-fat chow), and CoQ10 intervention group(45% high-fat chow+ 100 mg·kg -1·d -1CoQ10) based on their body weights according to the randomized block design. The body weight and food intake of mice in each group were collected. The levels of serum total cholesterol, triglyceride, low density lipoprotein-cholesterol, high density lipoprotein-cholesterol, alanine aminotransferase, and aspartate aminotransferase were detected. The contents of 17 bile acids in serum, liver, and colon contents of mice were detected by ultra-performance liquid chromatography-tandem mass(UPLC-MS/MS). The protein expressions of cholesterol 12α-hydroxylase(CYP8B1) and oxysterol 7α-hydroxylase(CYP7B1) in liver were detected by Western blotting. Results:CoQ10 significantly reduced body weight and ameliorated lipid metabolism disorders in mice fed a high-fat diet. Compared with the control group, serum total bile acid levels were reduced in the high-fat diet group( P<0.05); CoQ10 intervention elevated serum and colonic total bile acid levels( P=0.021, P=0.014) and increased liver, colon, and serum deoxycholic acid and ursodeoxycholic acid levels( P<0.05) in the mice compared with the high-fat diet group. Both colonic and serum deoxycholic acid levels in the CoQ10 intervention group were negatively correlated with body weights( P=0.024, P=0.019), and colonic deoxycholic acid and total cholesterol levels were also negatively correlated( P=0.006). CoQ10 increased the expression of CYP8B1 and CYP7B1 proteins in the liver of mice. Conclusion:CoQ10 can modulate bile acid metabolism in high-fat diet-fed mice and alleviate their obesity and lipid metabolism disorders.

Objective:To investigate the pathological characteristics and the clinical significance of novel coronavirus (2019-nCoV)-infected pneumonia (termed by WHO as coronavirus disease 2019, COVID-19).Methods:Minimally invasive autopsies from lung, heart, kidney, spleen, bone marrow, liver, pancreas, stomach, intestine, thyroid and skin were performed on three patients died of novel coronavirus pneumonia in Chongqing, China. Hematoxylin and eosin staining (HE), transmission electron microcopy, and histochemical staining were performed to investigate the pathological changes of indicated organs or tissues. Immunohistochemical staining was conducted to evaluate the infiltration of immune cells as well as the expression of 2019-nCoV proteins. Real time PCR was carried out to detect the RNA of 2019-nCoV.Results:Various damages were observed in the alveolar structure, with minor serous exudation and fibrin exudation. Hyaline membrane formation was observed in some alveoli. The infiltrated immune cells in alveoli were majorly macrophages and monocytes. Moderate multinucleated giant cells, minimal lymphocytes, eosinophils and neutrophils were also observed. Most of infiltrated lymphocytes were CD4-positive T cells. Significant proliferation of type Ⅱ alveolar epithelia and focal desquamation of alveolar epithelia were also indicated. The blood vessels of alveolar septum were congested, edematous and widened, with modest infiltration of monocytes and lymphocytes. Hyaline thrombi were found in a minority of microvessels. Focal hemorrhage in lung tissue, organization of exudates in some alveolar cavities, and pulmonary interstitial fibrosis were observed. Part of the bronchial epithelia were exfoliated. Coronavirus particles in bronchial mucosal epithelia and type Ⅱ alveolar epithelia were observed under electron microscope. Immunohistochemical staining showed that part of the alveolar epithelia and macrophages were positive for 2019-nCoV antigen. Real time PCR analyses identified positive signals for 2019-nCoV nucleic acid. Decreased numbers of lymphocyte, cell degeneration and necrosis were observed in spleen. Furthermore, degeneration and necrosis of parenchymal cells, formation of hyaline thrombus in small vessels, and pathological changes of chronic diseases were observed in other organs and tissues, while no evidence of coronavirus infection was observed in these organs.Conclusions:The lungs from novel coronavirus pneumonia patients manifest significant pathological lesions, including the alveolar exudative inflammation and interstitial inflammation, alveolar epithelium proliferation and hyaline membrane formation. While the 2019-nCoV is mainly distributed in lung, the infection also involves in the damages of heart, vessels, liver, kidney and other organs. Further studies are warranted to investigate the mechanism underlying pathological changes of this disease.

Objective To investigate the clinical value of serum lipoprotein associated phospholipase A 2 (Lp-PLA2)as a biomarker for benign prostatic hyperplasia(BPH).Methods A total of 65 serum samples of BPH patients were selected as BPH group,64 serum samples of healthy male as control group.The serum lev-el of Lp-PLA2,total prostate specific antigen(tPSA)and free prostate specific antigen(fPSA)in both groups were detected,and the specificity and sensitivity of Lp-PLA2,F/T ratio and combine both were analyzed by ROC curve.Results The serum level of Lp-PLA2,tPSA and fPSA in BPH group were significantly higher than those in control group,the different were significant(P<0.05).ROC curve analyze shown that the area under the curve(AUC)of Lp-PLA2 was 0.763,95% confidence interval(CI)was 0.680-0.833;AUC of F/T ratio was 0.715,95% CI 0.633 -0.795;AUC of Lp-PLA2 combined with F/T ratio was 0.832,95% CI 0.756-0.892.Conclusion The serum level of Lp-PLA2 could be used as a potential diagnostic marker for BPH,and the diagnostic value of Lp-PLA2 combined with F/T ratio was better than ther single indicator. Key words:benign prostatic hyperplasia; lipoprotein associated phospholipase A 2; total prostate spe-cific antigen; free prostate specific antigen; F/T ratio

Objective To investigate the correlation of the 1896 and 1899 mutations of hepatitis B virus (HBV)with the conversion of e antigen in serum and the progression of the disease. Methods 238 serum samples from the patients with HBsAg positive for over six months and HBV-DNA copy number > 5.0 × 102 IU/mL were collected,and the sequence analysis was used to analyze the nucleotide sequences of the 1896 and 1899 sites in the pre-C region of HBV. At the same time,the relevant clinical data and the expressions of HBeAg were collected,followed by Spearman correlation analysis and chi square test with SPSS 20.0. Results Both 1896 and 1899 sites in the pre-C region of HBV were mutated,and the base G was A,which was closely related to the expression of e antigen(P<0.05). Both G1896A and G1899A promoted the e antigen serological conversion ,and the e antigen serological conversion of G1899A was higher than that of G1896A. G1899A was associated with HBV related disease progression (correlation coefficient 0.280,P < 0.05),especially with the incurrence of HCC. Conclusions G1896A and G1899A in the pre-C region of HBV can promote the serological conversion of e antigen.

Objective To explore clinical results of posterior debridement combined with atlantoaxial fusion for upper cervical Tuberculosis.Methods From March 2007 to April 2012,8 patients with upper cervical Tuberculosis underwent posterior debridement combined with atlantoaxial fusion in our hospital were selected for retrospective analysis.3 cases were males and 5 females,aged 29-65 (43.5 ±13.2) years.According to the pedicle destruction,using different screws (pedicle screw or laminar screw)fixation.In the preoperative and final follow-up,Japanese Orthopaedic Association score (JOA) and neck disability index (NDI) were used to evaluate neurological function and calculate improvement rate JOA score.At final follow-up,clinical efficacy was evaluated by Odom's grade.situation of internal fixation,fusion of upper cervical were assessed by imaging examination.During follow-up,complications were documented and analyzed.Results Postoperatively 12 months,all bony fusion were achieved.Tuberculosis were reached clinical cure in 12-18 months.The JOA score increased from 10.5 ± 2.0 preoperatively to 15.6 ± 1.1 in final follow-up(P ＜0.05),and the NDI decreased from 29.9 ± 6.2 preoperatively to 8.6 ±1.6 (P ＜ 0.05).At last follow-up,according to Odom's standard,excellent were obtained in 6 cases(75.0％),good 1 cases (12.5％) and ordinary 1 case (12.5％).No severe complications was documented during follow-up.Conclusions The treatment of posterior debridement combine with atlantoaxial fusion,and structure grafting and local anti-Tuberculosis drug using intraoperative,not only could obtain reliable clinical efficacy,completely removal of lesions,but also obtain strong stability,which plays an important role in the treatment of cervical tuberculosis.

Objective To explore clinical results of posterior debridement combined with atlantoaxial fusion for upper cervical Tuberculosis.Methods From March 2007 to April 2012,8 patients with upper cervical Tuberculosis underwent posterior debridement combined with atlantoaxial fusion in our hospital were selected for retrospective analysis.3 cases were males and 5 females,aged 29-65 (43.5 ±13.2) years.According to the pedicle destruction,using different screws (pedicle screw or laminar screw)fixation.In the preoperative and final follow-up,Japanese Orthopaedic Association score (JOA) and neck disability index (NDI) were used to evaluate neurological function and calculate improvement rate JOA score.At final follow-up,clinical efficacy was evaluated by Odom's grade.situation of internal fixation,fusion of upper cervical were assessed by imaging examination.During follow-up,complications were documented and analyzed.Results Postoperatively 12 months,all bony fusion were achieved.Tuberculosis were reached clinical cure in 12-18 months.The JOA score increased from 10.5 ± 2.0 preoperatively to 15.6 ± 1.1 in final follow-up(P ＜0.05),and the NDI decreased from 29.9 ± 6.2 preoperatively to 8.6 ±1.6 (P ＜ 0.05).At last follow-up,according to Odom's standard,excellent were obtained in 6 cases(75.0％),good 1 cases (12.5％) and ordinary 1 case (12.5％).No severe complications was documented during follow-up.Conclusions The treatment of posterior debridement combine with atlantoaxial fusion,and structure grafting and local anti-Tuberculosis drug using intraoperative,not only could obtain reliable clinical efficacy,completely removal of lesions,but also obtain strong stability,which plays an important role in the treatment of cervical tuberculosis.

OBJECTIVETo explore the enhanced sensitivity of leukemia cell line KG-1a to activated immune cell-mediated cytolysis after treated with resveratrol.

METHODSThe value of 50% inhibition concentration (IC₅₀) for KG-1a by resveratrol was analyzed using trypan blue staining. Peripheral blood mononuclear cells were separated, and then activated by interleukin (IL)-2 and IL-15. The sensitivity of KG-1a treated with and without resveratrol to activated immune cell-mediated cytolysis was assayed by lactate dehydrogenase (LDH) -releasing assay. The expression of tumor necrosis factor related apoptosis inducing ligand (TRAIL) on the surface of activated immune cells and its receptors (DR4/5 and DcR1/2) on the surface of KG-1a were detected by flow cytometry.

RESULTSResveratrol could inhibit the proliferation of KG-1a and IC50 at 24 h was 25 mmol/L. At a ratio of 10:1 or 20:1 between effect and target, the cytolytic rates of treated KG-1a by activated immune cells were (55.80 ± 10.88)% and (72.31 ± 13.06)%, significantly higher than (24.96 ± 9.25)% and (37.93 ± 5.21)% of untreated KG-1a (P<0.05). The expression of DR5 on the surface of KG-1a treated with resveratrol was (9.05 ± 3.57)%, significantly higher than (3.11 ± 0.54)% of untreated KG-1a (P<0.05). Conversely, the expression of DcR1 on the surface of treated KG-1a was (13.23 ± 3.56)%, lower than (53.75 ± 10.51)% of KG-1a (P<0.05). When TRAIL pathway on the surface of activated immune cells was blocked, the cytolytic rates of treated KG-1a were (35.97 ± 6.36)% and (49.80 ± 10.68)%, significantly lower than (52.92 ± 6.98)% and (70.73 ± 9.79)% of untreated KG-1a (P<0.05) at the same ratio of effector and target.

CONCLUSIONResveratrol could enhance cytolytic sensitivity of KG-1a by activated immune cells through TRAIL pathway.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Humans ; Leukemia ; metabolism ; pathology ; Leukocytes, Mononuclear ; drug effects ; immunology ; metabolism ; Male ; Receptors, TNF-Related Apoptosis-Inducing Ligand ; metabolism ; Receptors, Tumor Necrosis Factor, Member 10c ; metabolism ; Stilbenes ; pharmacology ; TNF-Related Apoptosis-Inducing Ligand ; metabolism

Objective To evaluate the quality of life for tuberculosis patients after anterior debridement, autograft bone fusion and anterolateral fixation operation. Methods 17 cases of thoracolumbar spinal tuberculosis were treated surgically from January 2008 to March 2011. All the cases underwent anterior debridement, autograft bone fusion and anterolateral fixation operation. MOS health survey 36-item Short Form (SF-36), Japanese Orthopaedic Association Scores (JOA) and Visual Analogue Scale (VAS) were used to evaluate the quality of life, spine function and pain symptom before and 1 and 6 months after surgery. Results Compared to the results 1 month after surgery, the scores of physical function (PF), role physical (RP), bodily pain (BP), general health (GH), vitality (VT), social function (SF), role emotional (RE), and mental health (MH), and JOA subjective symptoms, JOA clinical signs, and JOA daily activity limitation, and VAS improved (P<0.05) 6 months after surgery; compared to pretreatment, the scores of PF, BP, GH, VT, SF, and MH, and JOA subjective symptoms, JOA clinical signs, and JOA daily activity limitation, and VAS improved (P<0.05). Conclusion The anterior debridement, autograft bone fusion and anterior fixation operation is effective to improve the quality of life, spine function and pain symptom for tuberculosis patients.

Objective:To investigate the therapeutic range of tacrolimus and effects of tacrolimus on liver and re- nal functions and blood routine in renal transplant recipients.Method:The whole blood tacrolimus concentration was meas- ured by micro-particle enzyme immunoassay(MEIA).Blood tacrolimus concentrations in 390 cases of renal transplant re- cipients were analyzed.The effects of tacrolimus on liver and renal function and blood routine were also studied.Result: The blood tacrolimus concentrations in 377 of 390 cases were within the range from 3 to 15?g?L~(-1).Their blood tacrolimus concentration differed greatly in renal transplant recipients within 6 months after transplantation.Their blood tacrolimus concentration was gradually decreased as time went on.Tacrolimus with therapeutic dosage had no effects on liver and renal function and blood routine.Conclusion:The therapeutic ranges of tacrolimus with MEIA were as follows:5 to 15?g?L~(-1) within 3 months after transplantation,5 to 10?g?L~(-1)between 4 to 6 months after transplantation,3 to 10?g?L~(-1)6 months after transplantation.The administration of tacrolimus had no effects on the liver and renal function and blood routine in re- nal transplant recipients.