Abstract: Objective　To analyze the nucleic acid detection results of severe acute respiratory syndrome corona virus-2 (SARS-CoV-2) by droplet digital PCR (ddPCR) and compare with the detection results of real-time fluorescence quantitative RT-PCR (qRT-PCR), so as to evaluate the advantages and disadvantages of detection, and to provide data support for optimizing the nucleic acid detection scheme of SARS-CoV-2. Methods　According to the SARS-CoV-2 specific primer probe published by the China Center for Disease Control and Prevention, a ddPCR detection method for SARS-CoV-2 was designed. One sample was selected for sensitivity test after gradient dilution; six respiratory virus nucleic acid positive samples including seasonal H3N2 influenza virus and SARS-CoV-2 positive samples were selected for specificity test; five SARS-CoV-2 positive samples were selected for repeatability test; in addition, 30 positive and 20 negative SARS-CoV-2 samples were selected for multiple clinical samples testing, and the results were analyzed and compared with those of qRT-PCR. Results　The ddPCR method can specifically detect SARS-CoV-2, and directly obtain the original copy number of the sample target gene to achieve accurate quantification; the sensitivity test of gradient dilution positive samples showed that qRT-PCR detected target genes in part of the 10-5 dilution of samples, and no target genes were detected in 10-6 dilution, while ddPCR detected all target genes in both 10-5 and 10-6 dilution of samples. The detection limit of ddPCR was two orders of magnitude higher than that of qRT-PCR, and the sensitivity was higher than that of qRT-PCR; in the comparison of the repeatability test results of the two methods, the coefficient of variation of ddPCR was 1.266%-11.814%, lower than 1.729%-26.174% of qRT PCR, and the repeatability was higher than qRT-PCR; among 50 clinical samples, 30 positive samples of confirmed cases of Coronavirus Disease 2019 (COVID-19) were detected by both methods, SARS-CoV-2 was successfully detected by both methods, and 20 negative samples of COVID-19 were detected by both methods, and the results were negative, with a coincidence rate of 100.00% (50/50). Conclusion　The ddPCR method can accurately quantify SARS-CoV-2 with strong specificity, and its sensitivity and repeatability are higher than those of qRT-PCR, but it also has certain detection limitations and is more suitable for the detection of low load samples. In the actual detection, the two methods can be reasonably combined to improve the detection accuracy.

Objective: This aim of the study is to analyze the general and specific inhibition control of college students who have mobile phone dependence and their craving for mobile phone after the wastage of inhibition control resource, and to provide a reference for the intervention. Methods: The Mobile Phone Addiction Tendency Scale was used to screen 40 mobile phone dependence and 40 non-dependence college students, who were divided into two groups completed two conditions of Numerical Stroop Task with and without mobile phone ringing sound interference. Craving for mobile phone before and after the Numerical Stroop Task was assessed. Results: Mobile phone dependence group had significantly more Stroop interference scores than mobile phone normal use group under the two conditions( F =19.34, P <0.01).After completing the Numerical Stroop Task under the interference condition, mobile phone dependence group experienced a significant boost craving of mobile phone ( t =-6.02, P <0.01). The scores of craving for mobile phones after the Numerical Stroop Task had significant positive relationships with Stroop interference scores( r =0.51, 0.51 , P <0.05). Conclusion The mobile phone dependence college students have poor inhibition control, inhibitory control which might associate with increase of craving for mobile phone.

Objective To investigate the effect of urate-lowering therapy on renal function in chronic kidney disease (CKD) stages 2-5 patients with hyperuricemia (HUA).Methods A total of 132 patients of CKD stages 2-5 with HUA between July 2016 and December 2017 in Department of Nephrology of the Second Affiliated Hospital of Anhui Medical University were prospectively and self-controlled analyzed.Serum uric acid (SUA),estimated glomerular filtration rate (eGFR) and other clinical parameters were measured at baseline and after 1-6 months treatment.The patients were divided into group A (CKD stages 2-3a) and group B (CKD stages 3b-5) on the baseline value of eGFR.The changes of SUA and eGFR before and after treatment were compared.According to the level of SUA after 6 months treatment,patients were divided into attainment group (SUA ＜ 360 μmol/L) and nonattainment group (SUA ≥ 360 μmol/L).The difference of renal function in pre-treatment and post-treatment was compared.Multiple stepwise linear regression was used to analyze the relationship among the change of eGFR after receiving 6 months' treatment (deGFR) and SUA level,baseline eGFR and other indexes.Results After 1,3,6 months treatment,the average levels of SUA,Scr and urea nitrogen of all patients were decreased significantly while eGFR value was increased significantly (all P ＜ 0.050) than those in pre-treatment period.After six-month-therapy,proteinuria and hematuria were improved significantly in all patients (P ＜ 0.001,P=0.001).Compared with pre-treatment period,both the SUA levels of group A and group B were declined significantly while eGFR had a significant rise after treatment (P ＜ 0.001).The change of eGFR post-treatment in group A was significantly higher than that of group B [(13.64±15.35) vs (8.97±9.79) ml· min-1· (1.73 m2)-1,P=0.044].At 6 months after treatment,the eGFR value increased markedly in both attainment group and nonattainment group compared with pre-treatment period (P ＜ 0.001).After six-month-therapy,the eGFR value in attainment group was increased more obviously than that of nonattainment group [(13.96 ± 14.64) vs (8.03±9.69) ml· min-1· (1.73 m2)-1,P=0.021].Multiple stepwise linear regression analysis showed that the baseline eGFR value was an influencing factor of deGFR (b=0.161,P=0.020).Conclusions The renal function of CKD stages 2-5 patients with HUA can be significantly improved by urate-lowering therapy,which can effectively reduce proteinuria and hematuria.

To investigate the clinic value of microsurgical shunt for the treatment of varicocele combined with asthenspermia,the clinical data and therapeutic method for 3 patients,who conducted the microscope spermatic vein high ligation combined with superficial epigastric vein flow,were retrospectively analyzed.No postoperative complications were found,and the original symptoms and signs were disappeared.All patients were conducted scrotal ultrasound and semen routine after 3 months,and all indexes,including maximum internal diameter of the cord vein (erect position),sperm density,sperm survival rate,sperm deformity rate and sperm forward movement rate,were gradually improved.Consequently,high ligation of spermatic vein combined with vascular bypass surgery under the microscope can block the countercurrent venous blood and establish a new return channel to the testis.Meanwhile,it can also protect the testicular artery and lymph-vessel.It is worth to be spread for the treatment ofvaricocele combined with asthenospermia.

This study was aimed to investigate the biological effects of Rhesus bone marrow mesenchymal stem cells (R-BMMSC) transfected by adenovirus bearing extracellular superoxide dismutase gene (AD-ECSOD). Using density gradient centrifugation and adherent culture way, the R-BMMSC transfected by AD-ECSOD and reporter gene EGFP were isolated, cultured and purified; the transfection efficiency was detected by fluorescence microscopy and flow cytometry; the ECSOD protein expression in cell culture supernatant were detected by ELISA; the surface antigens on R-BMMSC (CD34, CD29, CD45, CD90, HLA-DR) were detected by flow cytometry; and differentiation capability of transfected R-BMMSC were detected by oil red O and alizarin staining; the proliferation capability of R-BMMSC was assay by MTT method. The results showed that the transfection efficiency of AD-ECSOD (MOI 500, 1 000, 1 500 and 2 000) for R-BMMSC was > 95%. At 24 h after transfection, the ECSOD protein could be detected in cell culture supernatant, and its level was significantly higher than that of control group (P < 0.01). At 48 h after transfection, the expression level of ECSOD protein on MOI 1 500 and 2 000 was the highest. The proliferative capability, surface antigen expression and multi directive differentiation ability of transfected R-BMMSC were similar to non-transfected R-BMMSC. It is concluded that the AD-ECSOD can effectively transfect the R-BMMSC without influences on its biological features.
Adenoviridae ; genetics ; Adipogenesis ; Animals ; Bone Marrow Cells ; cytology ; Cell Differentiation ; Cells, Cultured ; Genetic Vectors ; Macaca mulatta ; Mesenchymal Stromal Cells ; cytology ; Osteoblasts ; cytology ; Superoxide Dismutase ; genetics ; Transfection

OBJECTIVETo detect the expression of thyroid transcription factor 1 (TTF-1) and study its application in the diagnosis of lung carcinomas.

METHODSOf 134 specimens from lung lobectomy, 105 were primary lung carcinomas including 76 non-small cell carcinomas (NSCLCs), 28 small cell lung cancers (SCLCs) and 1 complex carcinoma (SCLC and SCC), and 29 were metastatic carcinomas. Expression of TTF-1 was detected by immunohistochemistry. The expression level of TTF-1 was graded as, +:6% to 25% of tumor cells positive, ++:26% to 50%, +++:51% to 75%, and ++++:> 76%.

RESULTSThe positive nuclear immunoreactivity of TTF-1 was identified in 23 of 28 SCLCs (82.1%), but none in squamous cell cancer (SCC) (P < 0.001). The positive expression rate of TTF-1 in lung adenocarcinomas (ACs) was 73.8% (31/42). There was no correlation between TTF-1 expression and ACs differentiation or ACs subtypes (P > 0.05). All but one (thyroid follicular carcinoma) metastatic ACs were TTF-1-positive. Mesenchymal component and lymphoid or inflammatory cells were consistently TTF-1-negative.

CONCLUSIONA significant difference of TTF-1 expression may assist in distinguishing SCLC from SCC, lymphoma or inflammatory lesions. Owing to its restrictive expression in lung tissue, TTF-1 might be used to differentiate primary from metastatic adenocarcinoma of the lung.

Adenocarcinoma ; diagnosis ; metabolism ; Breast Neoplasms ; pathology ; Carcinoma, Non-Small-Cell Lung ; diagnosis ; metabolism ; Colorectal Neoplasms ; pathology ; Diagnosis, Differential ; Humans ; Lung Neoplasms ; diagnosis ; metabolism ; secondary ; Nuclear Proteins ; biosynthesis ; Thyroid Nuclear Factor 1 ; Transcription Factors ; biosynthesis

OBJECTIVETo discuss the clinicopathological features and prognostic factors of gastric lymphoma.

METHODS83 gastric lymphoma cases were analyzed retrospectively in accordance to the criteria of the new World Health Organization classification for neoplastic diseases of the hematopoietic and lymphoid tissues. The correlations between clinicopathological features, therapeutic measures and survival were discussed.

RESULTSThe age of patients ranged from 25 to 77, with a median of 52. The number of males were similar to that of females. There were no specific symptoms. The most common symptoms were stomach ache (60 cases, 72%) or discomfort. The duration of symptoms was often long and with a history of chronic gastric diseases (21 cases, 25%). 13 cases had multiple lesions in the gastrointestinal mucosa. 51 cases (61%) were accompanied by lymph node involvement. According to the new World Health Organization classification for neoplastic diseases of the hematopoietic and lymphoid tissues, 57 cases were extranodal marginal zone lymphoma of mucosa-associated lymphoid tissue (MALT)-type (MALT lymphoma), 23 were diffuse large B cell lymphoma accompanying MALT lymphoma, 2 were diffuse large B cell lymphoma and 1 was follicular lymphoma. Of all the cases, 31 were stage I E, 38 stage II E, 8 stage III E and 6 stage IV by the Ann Arbor staging system (1972). The total 5-year and 10-year survival rates were 77.8% and 70.1% respectively, with the mean survival time of 146 months. The 5-year and 10-year survival rates of MALT lymphoma were 77.4% and 72.3%, the 5-year and 10-year survival rates of diffuse large B cell lymphoma accompanying MALT lymphoma were 81.8% and 68.2%, the 5-year survival rate of diffuse large B cell lymphoma was 50.0%.

CONCLUSIONSThere are no specific symptoms in gastric lymphoma patients. Extranodal marginal zone lymphoma of MALT-type is the main histopathological type of gastric lymphoma, often accompanied by multiple mucosa involvement and also often accompanied by a history of chronic gastric disease. The lesion is usually localized for a long time, with a very good prognosis. Survival rate has a significant correlation with lymph node involvement and clinical stage. No correlations were found between the survival rates with age, gender, B symptoms, invasive depth of the wall of stomach, the size and range of the tumors or different therapeutic measures.

Adult ; Aged ; Chemotherapy, Adjuvant ; Female ; Follow-Up Studies ; Gastrectomy ; methods ; Humans ; Lymphatic Metastasis ; Lymphoma ; pathology ; surgery ; therapy ; Lymphoma, B-Cell ; pathology ; surgery ; therapy ; Lymphoma, B-Cell, Marginal Zone ; pathology ; surgery ; therapy ; Lymphoma, Large B-Cell, Diffuse ; pathology ; surgery ; therapy ; Male ; Middle Aged ; Neoplasm Staging ; Radiotherapy, Adjuvant ; Retrospective Studies ; Stomach Neoplasms ; pathology ; surgery ; therapy ; Survival Rate

Prostaglandin synthase (PGS) can catalyze the production of various types of prostaglandins and regulate the expression levels of related substances. The regulation mechanisms of the PGS gene are closely related with the occurrence and development of prostate diseases. However, few studies are reported on the regulation mechanisms of PGS in prostatic diseases, such as benign prostatic hyperplasia (BPH) and prostate cancer (PCa), or on the relationship between PGS gene regulation and prostate diseases. This review aims to analyze their correlation and provide some ideas for the prevention and control of BPH and PCa by intervention of the prostaglandin synthase regulatory pathway.
Gene Expression Regulation ; Humans ; Male ; Prostaglandin-Endoperoxide Synthases ; genetics ; physiology ; Prostatic Hyperplasia ; enzymology ; genetics ; prevention & control ; Prostatic Neoplasms ; enzymology ; genetics ; prevention & control

The role of (pro)rennin receptor (PRR) in cardiomyocytes of a heart failure (HF) rat model was studied. Spontaneously hypertensive rats (SHR) with HF (SHR-HF) or not were identified by two-dimensional (2-D) ultrasound. Age-matched Wistar Kyoto normotensive (WKY) rats were used as controls. PRR short hair RNA (sh-RNA) was injected into the heart of SHR-HF. Simultaneously SHR and controls received the same shRNA injection into the heart. Scramble shRNA was injected into the heart as controls. The expression of PRR mRNA and protein in cardiomyocytes was detected by using real-time PCR and Western blotting respectively. The heart function was evaluated by 2-D ultrasound, including eject fraction (EF%), fractional shortening (FS%), left ventricle thickness (LV), and inter-ventricular septal thickness (IVS). The number of apoptotic cardiomyocytes was counted by using flow cytometry. The results showed that the mRNA and protein expression levels of PRR were significantly higher in cardiomyocytes of SHR-HF group than in those of SHR group or control group. The apoptosis of myocytes in SHR-HF group was increased as compared with SHR group or control group. After knock-down of PRR with shRNA in SHR-HF group, the apoptosis of myocytes was reduced, resulting in the improved heart function. It was suggested that down-regulation of PRR might protect the heart from development of HF in SHR-HF by inhibiting the apoptosis of cardiomyocytes.
Animals ; Apoptosis ; genetics ; physiology ; Blotting, Western ; Chymosin ; metabolism ; Disease Models, Animal ; Enzyme Precursors ; metabolism ; Gene Expression ; Heart Failure ; genetics ; metabolism ; physiopathology ; Male ; Myocardium ; metabolism ; pathology ; Myocytes, Cardiac ; metabolism ; RNA Interference ; Rats ; Rats, Inbred SHR ; Rats, Inbred WKY ; Receptors, Cell Surface ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

The aim of this study was to investigate the effect of recombinant human granulocyte stimulating factor (rhG-CSF) on blood coagulation of beagles irradiated by 2.3 Gy neutron so as to provide new therapy for blood coagulation disorder after neutron irradiation. 10 beagles were exposed to 2.3 Gy neutron, and then randomly assigned into supportive care group and rhG-CSF-treated group. The rhG-CSF-treated cohorts were injected subcutaneously with rhG-CSF (10 µg/kg·d) beginning at the day of exposure for 21 consecutive days. Peripheral blood platelet counts were examined once every two days. In vitro platelet aggregation test, thromboelastography and blood clotting tetrachoric tests were also performed. The results indicated that the blood clotting system of irradiated dogs was in hypercoagulable state in the early days after 2.3 Gy neutron irradiation, and became hypocoagulable at crisis later and were mainly on intrinsic coagulation pathway. Blood fibrinogen increased markedly during the course of disease, while platelet counts and aggregation function were decreased remarkably. rhG-CSF administered daily could correct hypercoagulable state induced by 2.3 Gy neutron irradiation at the early time post exposure, shortened the thromboplastin generation time and clotting formation, down-regulated the abnormal high fibrinogen in blood, and improved platelet aggregation function. It is concluded that rhG-CSF can improve coagulation disorders of irradiated dogs.
Animals ; Blood Coagulation ; drug effects ; Bone Marrow ; radiation effects ; Dogs ; Granulocyte Colony-Stimulating Factor ; pharmacology ; therapeutic use ; Humans ; Leukocyte Count ; Neutron Diffraction ; Platelet Count ; Radiation Dosage ; Radiation Injuries, Experimental ; physiopathology ; Recombinant Proteins