This study aims to investigate the effects of Japanese encephalitis virus(JEV)on TLRs signaling pathway and its regulation of the secretion of inflammatory factors during the infection of testicular interstitial cells,In this study,the mRNA levels of TLR3,TLR7,TLR8,TRIF and MyD88 genes were detected by qPCR after 1 MOI dose of JEV was inoculated into testicular stro-mal cells at different time periods.Western blot assay was used to detect the expression levels of TLR3,TLR7,TRIF and MyD88 protein at 6 h after JEV infection,and ELISA was used to detect the expression levels of IL-1β,IL-6 and TNF-α at different time periods(6,12 and 24 h).The re-sults showed as follows:After 6 h of JEV infection,the mRNA levels of TLR3,TLR7,TRIF and MyD88 genes were significantly up-regulated(P＜0.05),and the mRNA levels of TLR8 genes were down-regulated(P＜0.05).Western blot results showed that the protein expressions of TLR3,TLR7,TRIF and MyD88 were significantly up-regulated when JEV infected testicular stromal cells for 6 h(P＜0.05),which was consistent with the corresponding mRNA transcription levels.There was no significant change in TLR8 protein expression.ELISA results showed that 6 h after JEV infection of testicular stromal cells,IL-6 was significantly increased(P＜0.01),and the expressions of IL-1β and TNF-α were not changed.TLR3,TLR7,TLR8,TRIF and MyD88 were si-lenced by siRNA,and the silenced cells were inoculated with JEV for 6 h,and IL-6 expression lev-els were detected by ELISA.The results showed that silenced TLR3,TLR7,TLR8,TRIF and MyD88 could significantly reduce the increase of IL-6 secretion induced by JEV infection(P＜0.05).These results indicated that JEV could induce the expression of inflammatory factor IL-6 by activating TLR3,TLR7 and TLR8 signaling pathway after infection of testicular stromal cells.This study provides a reference for further elucidating the mechanism of reproductive disorders caused by JEV infection.

Porcine circovirus type 2(PCV2)is the main pathogen causing porcine circovirus related diseases.PCV2 infection in pigs may lead to porcine dermatitis and nephrotic syndrome(PDNS)and weaned piglets multiple system failure syndrome(PMWS),etc.At present,the pathogenic mechanism is not fully understood.PCV2 is a single strand of negative link DNA,which can cause immune suppression in the body and lead to increased secondary susceptibility,which has a syner-gistic effect with various pig diseases and brings major economic losses to the pig industry.Al-though there are commercial vaccines,the prevention of vaccines has certain limitations and there is no effective drug treatment so far,an outbreak will threaten people's life and health and public safety,resulting in significant economic losses.In order to understand the latest progress of PCV2 escape mechanism and prevention and control,this paper summarizes the inhibition of interferon production,regulation of apoptosis,regulation of autophagy,regulation of pyroptosis and inflam-matory response,evasion of adaptive immune response,and prevention and control of PCV2,in or-der to provide new theoretical ideas for the research and prevention and control of PCV2.

[Purpose]To analyze the compliance and influencing factors of endoscopic screening in high-risk population of upper gastrointestinal cancer(UGC)in Chongqing Municipality.[Methods]Risk assessment of UGC was conducted among residents aged 40～74 years old in the areas covered by the Chongqing Urban Cancer Early Diagnosis and Treatment Program from 2012 to 2019.The residents with high risk of UGC were advised to receive free endoscopic screening in designated hospitals.The compliance and influencing factors of endoscopic screening among high-risk sub-jects were analyzed.[Results]There were 266 611 residents who completed the questionnaires and UGC risk assessment,among whom 48 000(18.00％)were assessed as high risk.A total of 9 162 high-risk individuals received the following endoscopic screening with a compliance rate of 19.09％.Multivariate Logistic regression analysis showed that residents aged 45～64 years old,with high school or above education,divorced or widowed status,occupational exposure to haza-rdous substances,hot food preference,high fat diet,frequent consumption of pickled and dried food,exposure to kitchen fume,psychic trauma or depression,upper gastrointestinal disease his-tory and family history of UGC were likely to accept endoscopic screening;while those aged 70 years old and above,current smokers,and having regular physical exercise were likely to have low compliance.[Conclusion]Among high-risk residents of UGC in Chongqing,the compliance to endoscopic screening needs be improved,health education and management should be targeted to those likely to have low compliance.

Objective    To summarize the clinical experiences of minimally invasive cardiac surgery (MICS) for cardiac atrioventricular valve reoperation. Methods    Perioperative data of 32 patients who underwent MICS for cardiac atrioventricular valve reoperation from 2009 to 2019 in the First Affiliated Hospital of Anhui Medical University were retrospectively reviewed, including 13 males and 19 females with a mean age of 51.0±12.6 years. All patients were given combined intravenous and inhalation anesthesia, and a double-lumen tube for mechanical ventilation. Cardiopulmonary bypass was established in all patients by femoral artery and venous cannulation or combined with percutaneous superior vena cava cannulation, without aortic cross-clamping. The MICS approaches included right anterolateral small incision surgery, thoracoscopic assisted small incision surgery and total thoracoscopic surgery. The clinical data of the 32 patients were compared with the perioperative indicators of 24 patients undergoing reoperation with conventional median thoracotomy during the same period. Results    Among them, 21 patients underwent isolated tricuspid valve replacement, 4 isolated tricuspid valvuloplasty, 1 combined tricuspid valve replacement and atrial septal defect repair and 6 combined mitral valve replacement and tricuspid valvuloplasty. Twenty-seven patients completed the operation in a beating heart, and 5 under the condition of ventricular fibrillation. Operation time (3.23±1.56 h vs. 5.46±2.13 h, P<0.001), postoperative mechanical ventilation time (9.19±5.40 h vs. 43.23±21.74 h, P<0.001), ICU stay (35.03±18.26 h vs. 79.15±22.43 h, P<0.001) and hospital stay of patients with minimally invasive surgery (9.35±6.43 d vs. 15.85±7.56 d, P=0.001) were shorter than those with median thoracotomy. And the extracorporeal circulation time was not significantly prolonged. There were 4 perioperative complications in patients with minimally invasive surgery, and 1 died in hospital after operation. Conclusion    MICS for cardiac atrioventricular valve reoperation can avoid the risk of median sternotomy and separation of cardiac scar adhesion. Especially, total thoracoscopic surgery has more advantages when compared with other operations, including less trauma, less myocardial ischemia reperfusion injury, more rapid recovery and fewer postoperative complications. Total thoracoscopic surgery may be the development direction of MICS for cardiac atrioventricular valve reoperation. However we should take effective and feasible measures to solve the problems caused by cardiopulmonary bypass.

PURPOSE: C-end rule (CendR) peptides are found to enhance the penetration of chemotherapeutic agents into tumor cells, while GX1 is a peptide that homes to gastric cancer (GC) vasculature. This study aimed to synthesize a novel peptide GX1-RPAKPAR (GXC) and to explore the effect of GXC on sensitizing GC cells to chemotherapeutic agents. MATERIALS AND METHODS: Intracellular Adriamycin concentration analysis was applied to conform whether GXC peptide increases the penetration of chemotherapeutic agents into GC cells in vitro. The effect of GXC peptide on sensitizing GC cells to chemotherapeutics was validated by apoptosis assay and in vitro/vivo drug sensitivity assay. The specificity of GXC to GC tissue was validated by ex vivo fluorescence imaging. RESULTS: In vitro, administration of GXC significantly increased Adriamycin concentrations inside SGC-7901 cells, and enhanced the efficacy of chemotherapeutic agents by decreasing the IC50 value. In vivo, FITC-GXC specifically accumulated in GC tissue. Moreover, systemic co-injection with GXC peptide and Adriamycin statistically improved the therapeutic efficacy in SGC-7901 xenograft models, surprisingly, without obviously increasing side effects. CONCLUSION: These results demonstrated that co-administration of the novel peptide GXC with chemotherapeutic agents may be a potential way to enhance the efficacy of anticancer drugs in GC treatment.
Apoptosis ; Doxorubicin ; Drug Therapy* ; Heterografts ; In Vitro Techniques ; Inhibitory Concentration 50 ; Optical Imaging ; Peptides ; Sensitivity and Specificity ; Stomach Neoplasms*

SUMMARY Thehumanembryonicstemcells(hESCs)serveasaself-renewable,genetically-healthy, pluripotent and single source of all body cells,tissues and organs.Therefore,it is considered as the good standard for all human stem cells by US,Europe and international authorities.In this study,the standard and healthy human mesenchymal progenitors,ligament tissues,cardiomyocytes,keratinocytes,primary neurons,fibroblasts,and salivary serous cells were differentiated from hESCs.The human cellular health-safety of NaF,retinoic acid,5-fluorouracil,dexamethasone,penicillin G,adriamycin,lead ace-tate PbAc,bisphenol A-biglycidyl methacrylate (Bis-GMA)were evaluated selectively on the standar-dized platforms of hESCs,hESCs-derived cardiomyocytes,keratinocytes,primary neurons,and fibro-blasts.The evaluations were compared with those on the currently most adopted cellular platforms.Parti-cularly,the sensitivity difference of PM2.5 toxicity on standardized and healthy hESCs derived fibroblasts, currently adopted immortalized human bronchial epithelial cells Beas-2B and human umbilical vein endo-thelial cells (HUVECs)were evaluated.The results showed that the standardized hESCs cellular plat-forms provided more sensitivity and accuracy for human cellular health-safety evaluation.

Objective:To differentiate human embryonic stem cells ( hESCs ) into keratinocytes ( K-hESCs) and analyse the expression characteristics of biomarkers of K-hESCs.Methods: The hESCs of line H9 were seeded on matrigel in mTeSR1 medium.The hESCs were directly differentiated into kerati-nocytes in epithelial differentiation medium with bone morphogenetic protein 4, retinoic acid and N2 sup-plement.The karyotype of K-hESCs was analyzed, comparing the gene expression differences of K-hESCs with human gingival epithelial cells (HGECs), human immortalized oral epithelial cells (HIOECs) and HaCaT by Real-time PCR.Molecular characteristics of the cell differentiation were observed throughout the process by immunocytochemical techniques.Results:H9-hESCs were successfully differentiated into the cells that exhibited characteristics of keratinocytes in epithelial differentiation medium.The karyotype of K-hESCs was 46, XX; and the keratinocyte gene p63 expression in K-hESCs was significantly lower than that in HaCaT ( P<0.05) , but there was no significant difference of p63 expression in K-hESCs, comparing with that in HGECs and HIOECs ( P >0.05 ) .Conclusion: H9-hESCs could be directly differentiated into K-hESCs.The gene expression of K-hESCs was similar to that of epithelial cells in the early stage of monolayer cells differentiation with high proliferative activity.

Objective To investigate the expression and clinical significance of serum microRNA (miRNA) expression profiling in the occurrence and progression of diabetic nephropathy.Methods The miRNA expression profiling was detected by miRNA TaqMan Low Density Array chip from 10 patient with diabetic nephropathy,10 diabetes patients with normoalbuminuria and 10 health control.Real-time quantitative PCR was applied to verify the result of miRNA array in serum samples of 66 patients with diabetic nephropathy (36 patients with microalbuminuria,30 patients with macroalbuminuria),40 diabetes patients with normoalbuminuria and 40 health control.And the relationship of differetial expression with clinical features was analyzed.Results miR-150-5p,miR-155-5p,miR-30e-5p and miR-3196 being validated by real-time quantitative PCR differentially expressed in 3 groups of serum samples from the diabetes patients with microalbuminuria (n=36),with normoalbuminuria (n=40) and health control (n=40) (P ＜ 0.05).Serum miR-150-5p (P=0.005) and miR-155-5p (P=0.006) changed significantly between diabetes patients with microalbuminuria (n=36) and with macroalbuminuria (n=30).Compared with the diabetes patients with microalbuminuria,serum miR-150-5p and miR-155-5p increased by 2.3 and 1.5 times in macroalbuminuria group,respectively.Estimated glomerular filtration rate and urinary albumin excretion rate significantly correlated with serum miR-150-5p and miR-155-5p level.Conclusions miR-150-5p and miR-155-5p may be involved in the process of pathological mechanisms of diabetic nephropathy.Serum miR-150-5p and miR-155-5p may be regarded as potential biomarkers to diagnosis the occurrence and development of diabetic nephropathy.

OBJECTIVEThis study aims to investigate the effects of mineral trioxide aggregate (MTA) and calcium hydroxide on proliferation and differentiation of human pulp cells from primary and permanent teeth.

METHODSCell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. The mRNA expression levels of dentinogenesis-related factors, including alkaline phosphatase (ALP) and dentin sialophosphoprotein (DSPP), and odontoclastogenesis-related factors, such as osteo- protegerin (OPG) and receptor activator of NF-κB ligand (RANKL), were determined by real time polymerase chain reac- tion (PCR).

RESULTSPrimary and permanent pulp cells treated with calcium hydroxide exhibited significantly lower proli- feration rates than the control cells (P<0.01). By contrast, the MTA-treated group showed significantly higher proliferation rates than the control group (P<0.01). Real time PCR results showed that calcium hydroxide-treated primary pulp cells exhi- bited significantly decreased ALP, DSPP, and OPG expression compared with the control group (P<0.01). Conversely, the MTA-treated group displayed significantly increased ALP, DSPP, and OPG expression (P<0.01). Calcium hydroxide-treated primary pulp cells also exhibited significantly upregulated RANKL expression (P < 0.01); by contrast, MTA-treated cells did not show any change in RANKL expression (P>0.05). Likewise, MTA-treated permanent pulp cells showed significantly upregulated ALP and DSPP expression (P < 0.01). However, the calcium hydroxide-treated group remained almost the same as the control group (P > 0.05). Neither MTA nor calcium hydroxide affected OPG and RANKL expression in per- manent pulp cells (P > 0.05).

CONCLUSIONMTA is more suitable as a pulp-capping agent, particularly in primary teeth, than calcium hydroxide.

Aluminum Compounds ; Calcium Compounds ; Calcium Hydroxide ; Cell Differentiation ; Cell Proliferation ; Dental Pulp ; Dentition, Permanent ; Drug Combinations ; Extracellular Matrix Proteins ; Humans ; Oxides ; Phosphoproteins ; Sialoglycoproteins ; Silicates

Objective This study aims to investigate the effects of mineral trioxide aggregate (MTA) and calcium hydroxide on proliferation and differentiation of human pulp cells from primary and permanent teeth. Methods Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) assay. The mRNA expression levels of dentinogenesis-related factors, including alkaline phosphatase (ALP) and dentin sialophosphoprotein (DSPP), and odontoclastogenesis-related factors, such as osteo-protegerin (OPG) and receptor activator of NF-κB ligand (RANKL), were determined by real time polymerase chain reac-tion (PCR). Results Primary and permanent pulp cells treated with calcium hydroxide exhibited significantly lower proli-feration rates than the control cells (P<0.01). By contrast, the MTA-treated group showed significantly higher proliferation rates than the control group (P<0.01). Real time PCR results showed that calcium hydroxide-treated primary pulp cells exhi-bited significantly decreased ALP, DSPP, and OPG expression compared with the control group (P<0.01). Conversely, the MTA-treated group displayed significantly increased ALP, DSPP, and OPG expression (P<0.01). Calcium hydroxide-treated primary pulp cells also exhibited significantly upregulated RANKL expression (P<0.01); by contrast, MTA-treated cells did not show any change in RANKL expression (P>0.05). Likewise, MTA-treated permanent pulp cells showed significantly upregulated ALP and DSPP expression (P<0.01). However, the calcium hydroxide-treated group remained almost the same as the control group (P>0.05). Neither MTA nor calcium hydroxide affected OPG and RANKL expression in per-manent pulp cells (P>0.05). Conclusion MTA is more suitable as a pulp-capping agent, particularly in primary
teeth, than calcium hydroxide.