This study aims to inquire about the fluctuations of ubiquitin-specific protease 47 on neu-roblastoma cells(Neuro-2a,N2a)infected by rabies virus(RABV).USP47 expression levels were detected after RABV infection in N2a cells through RT-qPCR,protein immunoblotting,and virus titer determination.The levels of RABV nucleoprotein and phosphoprotein gene and protein,and RABV titers in supernatants were analyzed during overexpression and knockdown of USP47.The results showed that RABV infection increased USP47 gene level in N2a cells.When overexpression of USP47,the levels of RABV N and P were increased,and the virus titers were also improved.Mo-reover,the level of interleukin-6(IL-6)genes decreased.Knocking down USP47 expression reduced levels of RABV N and P genes and proteins,lowered the virus titer,and elevated the IL-6 gene lev-el.The results suggest that USP47 promotes RABV infection and suppresses IL-6 expression.This finding lays the foundation for further investigation into the molecular mechanisms by which USP47 regulates RABV infection.

At present,some new tongue manifestations with distinct characteristics and high clinical value have not yet gained widespread consensus,which is unfavorable to their research and promotion.This paper representatively explores four types of tongue texture manifestations(liver gall line,bulging vein tongue,concave tip tongue,convex surface tongue)and two types of tongue coating manifestations(cracked tongue coating,white saliva line)among them.It is found that the liver gall line manifests as dark stagnation of various forms on the tongue surface,indicating liver qi stagnation and blood stasis as well as heat-toxin accumulation;the bulging vein tongue is characterized by clearly raised blood vessels on the tongue surface,suggesting blood stasis due to yang deficiency or cold congelation;the concave tip tongue is characterized by a depressed tip,indicating an abnormal tongue frenulum or malnutrition of heart,lung,and kidney;the convex surface tongue features a broad and thick tongue body with an overall"convex"shape,which is observed in healthy individuals or suggests qi movement disorder and spleen and stomach stagnation;the cracked tongue coating is characterized by cracks only on the tongue coating,indicating qi deficiency with disordered fluid or yin consumption due to intense heat;the white saliva line presents as strip-like white foam bands on the tongue margin,suggesting liver qi stagnation and dampness stagnancy due to spleen deficiency.No consensus exists regarding issues such as naming and description,reflecting the necessity of expanding and standardizing the theory of tongue diagnosis.This paper aims to emphasize the value and standardization of such new tongue manifestations,enrich the theoretical system of tongue diagnosis in traditional Chinese medicine,and provide novel insights and reference basis for clinical syndrome differentiation.

Objective To explore the ability of different regions of interest(ROI)location settings for the detection of bronchial artery and pulmonary vascular lesions in children with hemoptysis via computed tomography angiography(CTA)examination.Methods The hemoptysis group(79 cases)who underwent chest CTA examination and the control group(79 cases)with negative CTA results were retrospectively selected.The ROI of the hemoptysis group were placed in the pulmonary trunk,left ventricle,ascending aorta and descending aorta,which were defined as groups 1,2,3 and 4 respectively.Clinical data(age,sex)and CT parameters,including bronchial artery diameter,ascending aorta CT value,pulmonary artery CT value,△CTA-PA,bronchial artery image score,pulmonary artery image score,were recorded and analyzed,respectively.Results There was a statistically significant difference in the diameter of the left and right bronchial arteries between the hemoptysis group and the control group(P＜0.05).In the hemoptysis group,there were statistically significant differences in the image scores of the left and right bronchial arteries,pulmonary artery image scores,and△CTA-PA among the four subgroups(P＜0.05).According to post-hoc comparison results,there was a statistically significant difference in the overall mean scores of the right bronchial artery image score(x2=11.333,P＜0.05)and left bronchial artery image score(x2=8.111,P＜0.05)between groups 2 and 3.Conclusion When ROI is placed in the ascending aorta,bronchial artery lesions and pulmonary vascular lesions can be detected simultaneously in CTA examination,which is helpful for the diagnosis of hemoptysis vascular etiology in children.

This study investigates the feasibility of the VP8*protein as a subunit vaccine target for porcine rotavirus A(PoRVA),a major causative agent of diarrhea in piglets.The VP8* genes of PoRVA P[13]and P[23]genotype strains were amplified by RT-PCR.These genes were then liga-ted into the pET-28a(+)vector,yielding recombinant plasmids pET-28a-XJWF1-VP8*-P[23]and pET-28a-ShXYW13-VP8*-P[13].These plasmids were subsequently transformed into BL21(DE3)competent cells.The VP8*protein,induced by IPTG,was purified using affinity chroma-tography,and its expression and purification were verified by SDS-PAGE and Western blot.The purified VP8* protein was used to immunize mice,and serum samples were collected after three immunizations.Cross-neutralization assays were conducted to evaluate the ability of the VP8*protein immune serum to neutralize different genotype strains.The results demonstrated the ex-pression of soluble VP8*protein,with SDS-PAGE and Western blot analyses showing that the purified VP8*protein existed in both monomeric(27 kDa)and homodimeric(54 kDa)forms.ELISA results indicated that high levels of antibodies were produced in mice immunized with VP 8*-P[13]and VP8*-P[23]after three immunizations.Serum cross-neutralization assays revealed that the neutralizing titers of PoRVA VP8*-P[13]and VP8*-P[23]immune sera against homol-ogous genotype strains ranged from 1∶4 800 to 1∶19 200,significantly higher than those against heterologous genotype strains(1∶1 200).This suggests that the VP8*protein of different geno-type strains exhibits both antigenic conservation and distinct variability.The data obtained in this study provide a solid foundation for further exploration of the antigenic structure of the PoRVA VP8* protein and the development of novel subunit vaccines.

This study investigates the feasibility of the VP8*protein as a subunit vaccine target for porcine rotavirus A(PoRVA),a major causative agent of diarrhea in piglets.The VP8* genes of PoRVA P[13]and P[23]genotype strains were amplified by RT-PCR.These genes were then liga-ted into the pET-28a(+)vector,yielding recombinant plasmids pET-28a-XJWF1-VP8*-P[23]and pET-28a-ShXYW13-VP8*-P[13].These plasmids were subsequently transformed into BL21(DE3)competent cells.The VP8*protein,induced by IPTG,was purified using affinity chroma-tography,and its expression and purification were verified by SDS-PAGE and Western blot.The purified VP8* protein was used to immunize mice,and serum samples were collected after three immunizations.Cross-neutralization assays were conducted to evaluate the ability of the VP8*protein immune serum to neutralize different genotype strains.The results demonstrated the ex-pression of soluble VP8*protein,with SDS-PAGE and Western blot analyses showing that the purified VP8*protein existed in both monomeric(27 kDa)and homodimeric(54 kDa)forms.ELISA results indicated that high levels of antibodies were produced in mice immunized with VP 8*-P[13]and VP8*-P[23]after three immunizations.Serum cross-neutralization assays revealed that the neutralizing titers of PoRVA VP8*-P[13]and VP8*-P[23]immune sera against homol-ogous genotype strains ranged from 1∶4 800 to 1∶19 200,significantly higher than those against heterologous genotype strains(1∶1 200).This suggests that the VP8*protein of different geno-type strains exhibits both antigenic conservation and distinct variability.The data obtained in this study provide a solid foundation for further exploration of the antigenic structure of the PoRVA VP8* protein and the development of novel subunit vaccines.

At present,some new tongue manifestations with distinct characteristics and high clinical value have not yet gained widespread consensus,which is unfavorable to their research and promotion.This paper representatively explores four types of tongue texture manifestations(liver gall line,bulging vein tongue,concave tip tongue,convex surface tongue)and two types of tongue coating manifestations(cracked tongue coating,white saliva line)among them.It is found that the liver gall line manifests as dark stagnation of various forms on the tongue surface,indicating liver qi stagnation and blood stasis as well as heat-toxin accumulation;the bulging vein tongue is characterized by clearly raised blood vessels on the tongue surface,suggesting blood stasis due to yang deficiency or cold congelation;the concave tip tongue is characterized by a depressed tip,indicating an abnormal tongue frenulum or malnutrition of heart,lung,and kidney;the convex surface tongue features a broad and thick tongue body with an overall"convex"shape,which is observed in healthy individuals or suggests qi movement disorder and spleen and stomach stagnation;the cracked tongue coating is characterized by cracks only on the tongue coating,indicating qi deficiency with disordered fluid or yin consumption due to intense heat;the white saliva line presents as strip-like white foam bands on the tongue margin,suggesting liver qi stagnation and dampness stagnancy due to spleen deficiency.No consensus exists regarding issues such as naming and description,reflecting the necessity of expanding and standardizing the theory of tongue diagnosis.This paper aims to emphasize the value and standardization of such new tongue manifestations,enrich the theoretical system of tongue diagnosis in traditional Chinese medicine,and provide novel insights and reference basis for clinical syndrome differentiation.

Objective To explore the ability of different regions of interest(ROI)location settings for the detection of bronchial artery and pulmonary vascular lesions in children with hemoptysis via computed tomography angiography(CTA)examination.Methods The hemoptysis group(79 cases)who underwent chest CTA examination and the control group(79 cases)with negative CTA results were retrospectively selected.The ROI of the hemoptysis group were placed in the pulmonary trunk,left ventricle,ascending aorta and descending aorta,which were defined as groups 1,2,3 and 4 respectively.Clinical data(age,sex)and CT parameters,including bronchial artery diameter,ascending aorta CT value,pulmonary artery CT value,△CTA-PA,bronchial artery image score,pulmonary artery image score,were recorded and analyzed,respectively.Results There was a statistically significant difference in the diameter of the left and right bronchial arteries between the hemoptysis group and the control group(P＜0.05).In the hemoptysis group,there were statistically significant differences in the image scores of the left and right bronchial arteries,pulmonary artery image scores,and△CTA-PA among the four subgroups(P＜0.05).According to post-hoc comparison results,there was a statistically significant difference in the overall mean scores of the right bronchial artery image score(x2=11.333,P＜0.05)and left bronchial artery image score(x2=8.111,P＜0.05)between groups 2 and 3.Conclusion When ROI is placed in the ascending aorta,bronchial artery lesions and pulmonary vascular lesions can be detected simultaneously in CTA examination,which is helpful for the diagnosis of hemoptysis vascular etiology in children.

This study aims to inquire about the fluctuations of ubiquitin-specific protease 47 on neu-roblastoma cells(Neuro-2a,N2a)infected by rabies virus(RABV).USP47 expression levels were detected after RABV infection in N2a cells through RT-qPCR,protein immunoblotting,and virus titer determination.The levels of RABV nucleoprotein and phosphoprotein gene and protein,and RABV titers in supernatants were analyzed during overexpression and knockdown of USP47.The results showed that RABV infection increased USP47 gene level in N2a cells.When overexpression of USP47,the levels of RABV N and P were increased,and the virus titers were also improved.Mo-reover,the level of interleukin-6(IL-6)genes decreased.Knocking down USP47 expression reduced levels of RABV N and P genes and proteins,lowered the virus titer,and elevated the IL-6 gene lev-el.The results suggest that USP47 promotes RABV infection and suppresses IL-6 expression.This finding lays the foundation for further investigation into the molecular mechanisms by which USP47 regulates RABV infection.

In November 2018, the U.S. food and drug administration (FDA) issued guidance for the development of drugs for chronic hepatitis B virus infection (draft for comments) (hereinafter referred to as draft for comments), and in April 2022, the FDA issued Chronic Hepatitis B Virus Infection: Developing Drugs for Treatment, which has been updated with some details based on the Draft for Comments. This guidance further emphasizes the importance of HBsAg clearance in clinical trials, and classifies chronic suppressive therapy into two categories, namely noninferiority (NI) (or superiority) test with nucleos(t)ide analogues as control and add-on superiority trial with nucleos(t)ide analogues as control, and as for the latter, HBV DNA is no longer recommended as a primary endpoint of the trial, which poses a huge challenge to the development of innovative drugs targeting HBV DNA. The new finite duration therapy should aim to eliminate HBsAg and reduce virologic relapse and the risk of liver disease progression during treatment cessation. Reduction in HBsAg from baseline is not recommended as a primary endpoint for phase Ⅲ clinical trials, since the correlation between such reduction and clinical response remains unclear. In addition, this guidance also specifies the duration of treatment cessation and treatment consolidation period and the criteria for withdrawal of nucleos(t)ide analogues.

Objective: To investigate the rationale for appropriate diagnostic methods and treatment protocols for unexpected gallbladder carcinoma(UGC). Methods: The clinical and pathological data of 45 patients with UGC admitted at Department of General Surgery, Xin Hua Hospital, Shanghai Jiao Tong University School of Medicine,from January 2008 to December 2017 were retrospectively collected and analyzed.There were 11 males(28.9%) and 34 females(71.1%),aged 68 years(range:27 to 68 years).And there were 20 cases who aged above 70 years. Twenty-four cases were diagnosed preoperatively as cholecystolithiasis plus chronic cholecystitis.Ten cases were diagnosed preoperatively as cholecystolithiasis plus actue cholecystitis.Six cases were diagnosed preoperatively as cholecystolithiasis plus choledocholith.Six cases were admitted because of gallbladder polyp and 1 case was admitted because of gallbladder adenomyomatosis. Results: Thirty-four patients with UGC received radical surgery.Among them,11 patients experienced postoperative complication and no posterative mortality occoured during hospital stay.Thirteen patients were diagnosed with T1b UGC, the harvested lymph node of Nx, N0, N1 and N2 was 2, 9, 1 and 1, respectively.In addition, 2 cases were identified to have local-regional tumor recurrence during our rescue radical surgery.The median overall survival time of the patients who did not receive radical surgery was 7 months(range:2-56 months).Nevertheless,the median overall survival time for patients diagnosed with T1, T2 and T3 tumors who received radical surgery, was 41 months(range: 19-82 months), 33.5 months(range: 31-36 months) and 17 months(range: 7-46 months), respectively. Conclusions For patients with UGC, rescue radical surgery can achieve a better survival time.Furhtermore, our experience proved that rescue radical surgery for UGC is safe and feasible.Therefore,rescue radical surgery should be performed in patients with diagnose with UGC especially those T1b patients.