Objective: To prepare an inactivated hepatitis A vaccine using a attenuated strain of hepatitis A virus (HAV) H2 and to analyze its immunizing dose, so as to provide the reference for development and production of inactivated hepatitis A vaccines. Methods: Human embryonic lung diploid cells (KMB17) were infected with attenuated HAV H2 strain to proliferate the virus, then the cells containing viruses were harvested, extracted and purified. The obtained virus concentrate was prepared into vaccine bulk and test vaccines with 1 280 EU/mL antigen content. Vaccine testing was carried out according to the inactivated hepatitis A vaccine standards specified in the Part Ⅲ of the Pharmacopoeia of the People's Republic of China (2020 edition). A total of 110 mice were randomly divided into 11 groups, including 5 dose groups (80, 160, 320, 640 and 1 280 EU/dose) of the test vaccine and the reference vaccine, as well as the adjuvant control group. Mice were immunized twice by intraperitoneal injection, their serum HAV antibodies were detected, and the geometric mean titer (GMT) and positive conversion rate of antibodies were analyzed to evaluate the immunising dose of the vaccine. Results: The antigen content and viral titer of the virus harvest solution were 5 120 EU/mL and 8.33 lgCCID50/mL, respectively. The removal rate of foreign protein reached 98.05% and the recovery rate of antigen was 66.25%. The test vaccine met the requirements of Part Ⅲ of the Pharmacopoeia of the People's Republic of China (2020 edition). The GMTs of HAV antibodies in the test vaccine and the reference vaccine dose groups after the second immunization were more than twice higher than those after the first immunization. Regardless of primary immunization or secondary immunization, the GMTs (log2) of HAV antibodies in the test vaccine groups with doses of 160 EU/dose and above were higher than those in the 80 EU/dose group (all P<0.05), while there was no statistically significant differences between the dose groups of 160 EU/dose and above (all P>0.05). The antibody positive conversion rate of 160 EU/dose and above of the test vaccine was 100.00% after the secondary immunization. Conclusions The inactivated hepatitis A vaccine of attenuated H2 strain tested in this study demonstrates strong immunogenicity in mice, suggesting its potential as a candidate vaccine. The preliminary analysis indicates an immunizing dose of 320 EU/dose for children and 640 EU/dose for adults.

Objective:To analyze the etiology of chronic spontaneous urticaria (CSU) in children and associated factors affecting the disease severity.Methods:A single-center cross-sectional study was conducted. Children aged ≤ 17 years with CSU were prospectively enrolled at the Department of Dermatology, Children′s Hospital of Chongqing Medical University from November 2021 to November 2022. Clinical data were collected, serum total IgE and allergen-specific IgE (sIgE) were detected, and basophil activation test (BAT) and autologous serum skin test (ASST) were performed. According to the ASST and BAT results, the children were divided into the chronic autoimmune urticaria (CAU) group (positive for both ASST and BAT), non-CAU group (negative for both ASST and BAT), and partial CAU group (positive for either ASST or BAT). Differences in the etiology and clinical characteristics were analyzed between the CAU group and the non-CAU group. Based on the weekly urticaria activity score (UAS7), the children with CSU were divided into the mild group (UAS7 < 16 points) and moderate to severe group (UAS7 ≥ 16 points). Factors associated with the severity of CSU in children were analyzed using logistic regression. Non-normally distributed quantitative data were expressed as M ( Q1, Q3), and the non-parametric rank sum test (Kruskal-Wallis test) was used to compare quantitative data among multiple groups. Results:This study enrolled a total of 93 children with CSU, including 50 males (53.8%) and 43 females (46.2%), with the age being 5.9 (2.9, 9.2) years, and the disease duration being 4 (2, 8) months; 32 patients (34.4%) were complicated by angioedema, 28 (30.1%) had a family history of chronic urticaria, 49 (52.7%) had a family history of atopic diseases, 14 (15.1%) had a family history of autoimmune diseases, and 26 (28.0%) had at least one atopic comorbidity. Etiologic analysis showed that 32 cases (32/69, 46.4%) were positive for ASST and 28 (28/70, 40.0%) were positive for BAT. Both ASST and BAT were performed in 57 cases, and they were divided into the CAU group (18 cases), non-CAU group (24 cases), and partial CAU group (15 cases) according to the test results. There were no significant differences in the age, disease duration, gender ratio, proportion of patients with atopic comorbidity, or proportion of patients having a family history of atopic diseases among the 3 groups (all P > 0.05), while the proportion of patients with moderate to severe CSU (UAS7 ≥ 16 points) was higher in the CAU group (16/18) than in the non-CAU group (11/24, P < 0.05). Triggering factors were identified in 19 cases (20.4%), including 18 (19.3%) cases of food allergy and 1 case (1.0%) of antibiotic allergy. The serum total IgE level was elevated in 22 cases (22/89, 24.7%), and 40 (40/81, 49.4%) showed elevated levels of at least 1 sIgE. The UAS7 of the children with CSU was 16 (15, 21) points, and there were 31 (33.3%) children with mild CSU and 62 (66.7%) with moderate to severe CSU. Univariate logistic regression analysis showed that BAT positivity was associated with disease severity ( OR = 7.566, 95% CI: 2.238 - 25.572, P < 0.05). After adjustment for age and gender, multivariate logistic regression analysis showed that BAT positivity was associated with moderate to severe CSU ( OR = 6.725, 95% CI: 1.361 - 33.227, P < 0.05) . Conclusions:Autoimmunity may be the main cause of CSU in children, followed by allergic factors. ASST could be used as a primary screening test for the diagnosis of CAU in children, and BAT may help identify CAU and predict disease severity.

Background::The potential impact of pre-existing coronary artery stenosis (CAS) on acute pulmonary embolism (PE) episodes remains underexplored. This study aimed to investigate the association between pre-existing CAS and the elevation of high-sensitivity cardiac troponin I (hs-cTnI) levels in patients with PE.Methods::In this multicenter, prospective case-control study, 88 cases and 163 controls matched for age, sex, and study center were enrolled. Cases were patients with PE with elevated hs-cTnI. Controls were patients with PE with normal hs-cTnI. Coronary artery assessment utilized coronary computed tomographic angiography or invasive coronary angiography. CAS was defined as ≥50% stenosis of the lumen diameter in any coronary vessel >2.0 mm in diameter. Conditional logistic regression was used to evaluate the association between CAS and hs-cTnI elevation.Results::The percentage of CAS was higher in the case group compared to the control group (44.3% [39/88] vs. 30.1% [49/163]; P = 0.024). In multivariable conditional logistic regression model 1, CAS (adjusted odds ratio [OR], 2.680; 95% confidence interval [CI], 1.243–5.779), heart rate >75 beats/min (OR, 2.306; 95% CI, 1.056–5.036) and N-terminal pro-B type natriuretic peptide (NT-proBNP) >420 pg/mL (OR, 12.169; 95% CI, 4.792–30.900) were independently associated with elevated hs-cTnI. In model 2, right CAS (OR, 3.615; 95% CI, 1.467–8.909) and NT-proBNP >420 pg/mL (OR, 13.890; 95% CI, 5.288–36.484) were independently associated with elevated hs-cTnI. Conclusions::CAS was independently associated with myocardial injury in patients with PE. Vigilance towards CAS is warranted in patients with PE with elevated cardiac troponin levels.

OBJECTIVE: To explore the genetic basis of four children with congenital hyperinsulinemia (CHI). METHODS: The four children were subjected to high-throughput whole exome sequencing (WES). Candidate variants were validated by Sanger sequencing. RESULTS: WES analysis has identified 4 variants in the ABCC8 gene and 1 variant in GLUD1, including a ABCC8 c.382G>A variant in case 1, compound heterozygous c.698T>C and c.4213G>A variants of the ABCC8 gene concomitant with a de novo 14.9 Mb microduplication of chromosome 15 in case 2, and ABCC8 c.331G>A variant in case 3, and de novo c.955T>C variant of the GLUD1 gene in case 4. Of these, c.698T>C of the ABCC8 gene and c.955T>C of the GLUD1 gene were unreported previously. Based on the American College of Medical Genetics and Genomics guidelines, the c.382G>A(p.Glu128Lys), c.698T>C(p.Met233Thr) and c.4213G>A(p.Asp1405Asn) variants of ABCC8 gene and c.955T>C(p.Tyr319His) variant of GLUD1 gene were predicted to be likely pathogenic(PM1+PM2+PP3+PP4, PM1+PM2+PM5+PP3+PP4, PM1+PM2+PP3+PP4 and PS1+PM1+PM2+PP3), and the c.331G>A (p.Gly111Arg) variant of ABCC8 gene was predicted to be uncertain significance(PM1+PM2+PP4). CONCLUSION The variants of the ABCC8 and GLUD1 genes probably underlay the pathogenesis of CHI in the four patients. Above results have facilitated clinical diagnosis and genetic counseling for the affected families.
Child ; Genomics ; High-Throughput Nucleotide Sequencing ; Humans ; Hyperinsulinism ; Mutation ; Whole Exome Sequencing

OBJECTIVE: To analyze the metabolic profile and genetic variants for newborns with primary carnitine deficiency (PCD) from Guangxi, China. METHODS: From January 2014 to December 2019, 400 575 newborns from the jurisdiction of Guangxi Zhuang Autonomous Region Newborn Screening Center were subjected to tandem mass spectrometry (MS/MS) analysis. Newborns with positive results for PCD and their mothers were recalled for retesting. Those who were still positive were subjected to sequencing of the SLC22A5 gene. RESULTS: Twenty-two newborns and 9 mothers were diagnosed with PCD, which gave a prevalence rate of 1/18 208. Sequencing of 18 newborns and 4 mothers have identified 14 types of SLC22A5 gene variants, with the common ones including c.51C>G (10/44, 22.7%), c.1195C>T (9/44, 20.5%) and c.1400C>G (7/44, 15.9%), The c.517delC(p.L173Cfs*3) and c.1031C>T(p.T344I) were unreported previously and predicted to be pathogenic (PVS1+PM2_supporting+PM3+PP4) and likely pathogenic (PM1+PM2_supporting+PM3+PP3+PP4) based on the American College of Medical Genetics and Genomics standards and guidelines. CONCLUSION c.51C>G, c.1195C>T and c.1400C>G are the most common variants underlying PCD in Guangxi.
Cardiomyopathies ; Carnitine/deficiency* ; China ; Humans ; Hyperammonemia ; Infant, Newborn ; Metabolome ; Muscular Diseases ; Mutation ; Solute Carrier Family 22 Member 5/genetics* ; Tandem Mass Spectrometry

In April 2016, the Global Polio Eradication Initiative (GPEI) adjusted its polio vaccination strategy, converting trivalent oral polio vaccine (tOPV) into bivalent oral polio vaccine (bOPV), and withdrawing type 2 oral polio vaccine (OPV2) globally. However, after the withdrawal of OPV2, there were many outbreaks of type-2 circulating vaccine-derived poliovirus (cVDPV2) in Asia and Africa. In order to eradicate poliovirus completely, GPEI launched the research and development of the novel serotype 2 oral polio vaccine (nOPV2) in 2010 and considering whether it is necessary to reuse OPV. This paper summarizes the epidemiological situation of cVDPV2 before and after OPV2′s withdrawal, the related factors affecting the reuse of OPV and the related research progress of nOPV2.

Enteroviruses (EV) are the most common pathogens in humans, often causing large-scale infectious diseases, such as: hand, foot and mouth disease, herpes angina, myocarditis, encephalitis, aseptic meningitis, acute flaccidity Paralysis and acute flaccid myelitis and other nervous system and cardiopulmonary diseases, and them often infect children under 5 years old, severely can cause fatal complications. In recent years, the prevalence of non-enteric virus A71 (EV71) and non-coxsackievirus A16 (CV-A16) enteroviruses has gradually increased, and the dominant strains of EVs have gradually changed. A timely grasp of the etiology, epidemiology, and molecular evolution characteristics of EVs is of great significance to the prevention and control of EVs. Therefore, this article reviews the current status of diseases caused by non-EV71 and non-CV-A16 enteroviruses and analysis the molecular epidemiology, in order to have a certain prompting effect on the prevention and control of EVs.

In April 2016, the Global Polio Eradication Initiative (GPEI) adjusted its polio vaccination strategy, converting trivalent oral polio vaccine (tOPV) into bivalent oral polio vaccine (bOPV), and withdrawing type 2 oral polio vaccine (OPV2) globally. However, after the withdrawal of OPV2, there were many outbreaks of type-2 circulating vaccine-derived poliovirus (cVDPV2) in Asia and Africa. In order to eradicate poliovirus completely, GPEI launched the research and development of the novel serotype 2 oral polio vaccine (nOPV2) in 2010 and considering whether it is necessary to reuse OPV. This paper summarizes the epidemiological situation of cVDPV2 before and after OPV2′s withdrawal, the related factors affecting the reuse of OPV and the related research progress of nOPV2.

Enteroviruses (EV) are the most common pathogens in humans, often causing large-scale infectious diseases, such as: hand, foot and mouth disease, herpes angina, myocarditis, encephalitis, aseptic meningitis, acute flaccidity Paralysis and acute flaccid myelitis and other nervous system and cardiopulmonary diseases, and them often infect children under 5 years old, severely can cause fatal complications. In recent years, the prevalence of non-enteric virus A71 (EV71) and non-coxsackievirus A16 (CV-A16) enteroviruses has gradually increased, and the dominant strains of EVs have gradually changed. A timely grasp of the etiology, epidemiology, and molecular evolution characteristics of EVs is of great significance to the prevention and control of EVs. Therefore, this article reviews the current status of diseases caused by non-EV71 and non-CV-A16 enteroviruses and analysis the molecular epidemiology, in order to have a certain prompting effect on the prevention and control of EVs.

OBJECTIVE：To study the skin irritation and se nsitization of domestic generic drug Clobetasone butyrate cream ， and to compare it with commercial drug （original drugs ）. METHODS ：The skin irritation test was conducted on rabbits. Totally 24 rabbits were randomly divided into test preparation intact skin group ，test preparation abraded skin group ，commercial drug intact skin group and commercial drug abraded skin group ，with 6 rabbits in each group. 0.5 mL test preparation or commercial drug was administered to the left side of intact or abraded skin and the same amount of excipient on the right side of each rabbit twice a day for consecutive 7 days. The irritation of the drug to the rabbit skin was observed ，and the erythema and edema of the skin were scored；the skin of administration site was taken at 72 h after last administration and the end of 7 d after drug withdrawal for histopathological examination. The skin sensitization test （Buehler test ）was carried out on guinea pigs. Totally 60 guinea pigs were randomly divided into test preparation group （n＝20），commercial drug group （n＝20），positive control group （n＝10）and excipient control group （n＝10）. 0.2 mL test preparation or commercial drug was administrated to the left side of the rib abdomen skin of each guinea pig at the 0，7th，14th day to induce model ，and an equal amount of corresponding preparation was administered to the right side in the same way at the 28th day for stimulation. Hypersensitive response such as erythema and edema were observed and scored at 24 h and 48 h after the stimulation. The incidence of hypersensitive response was then calculated. RESULTS：In skin irritation test of rabbits ，no erythema and edema was caused by the test preparation or commercial drug on intact skin of rabbits ；scores of skin irritation was 0；there was no dermal irritation. Both test preparation and commercial drug caused transient slight erythema on abraded skin of a few rabbits ；scores of intact and abraded skin irritation were 0-0.33；there was no dermal irritation. There was no statistical significance among groups. No dermal pathological changes were observed. In skin sensitization test of guinea pig ，no hypersensitive response such as erythema and edema was found on the skin of guinea pigs in both test preparation and commercial drug groups ；both score and the incidence of hypersensitive response were 0. Compared with excipient control group ，there was no statistical significance of average score and the incidence of hypersensitive response in test preparation group and commercial drug group. CONCLU- SIONS：In skin irritation test of rabbits and skin sensitization test of guinea pigs ， the evaluation results of generic Clobetasone butyrate cream are the same as those of the original drug. It has no irritation to the skin of rabbit ，and no sensitization to the skin of guinea pigs.