Objective To report two cases of Candida parapsilosis-caused candidiasis characterized by verrucous nodules and masses,and to assess their clinical features,diagnosis and treatment.Methods A medical history including medication history and therapeutic response was carefully collected from two male patients.Physical examination was carried out with a focus on skin lesions.Diagnosis was made according to medical history as well as physical,mycological and histopathological examination findings.Antifungal agents were given at a high enough dose in time.After lesions improved,the doses of antifungal agents were tapered gradually,and drugs were withdrawn until patients completely healed.Compound preparations containing both antifungal agents and glucocorticoids were also topically applied in the early period of treatment.Results Both patients were diagnosed as Candida parapsilosis-caused candidiasis characterized by verrucous nodules and masses,and cured after 100-day supporting treatment and antifungal treatment with oral itraconazole,etc.Conclusions Candida parapsilosis-caused candidiasis should be managed with combination therapy mainly including antifungal agents.To achieve a satisfying efficacy,antifungal agents should be given early at a high enough dose for a long enough period.

Objective To observe therapeutic effects of acupoint catgut embedding therapy on ulcerative colitis.Methods 55 cases of ulcerative colitis were randomly recruited into a treatment group and a control group. The treatment group was treated with catgut embedding, and the control group was treated with oral administration of salicylazosul fapyridine. Results The therapeutic effect of the treatment group was better than those of the control group (P=0.002＜0.05). Conclusion Acupoint catgut embedding therapy has a better therapeutic effect on ulcerative colitis.

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 clone the mature peptide gene of seo, construct two kinds of prokaryotic fusion expression vector, obtain recombinant staphylococcal enterotoxin O(SEO) and evaluate its biological activity. Methods The seo gene was amplified from ZNZ2-3 strain of Staphylococcus xylosus by designed primers and it was cloned into fusion vector pGEX-6P-1 and pET28a. The soluble recombinant protein(GST-SEO) was expressed in E. coli BL21 host cells, the purified GST-SEO was obtained by a single step of affini-ty chromatography using Glutathinione Sepharose 4B colume. Another fusion protein(P28-SEO) with his-tag was expressed in E. coli BL21 ( DE3), the purified P28-SEO protein was harvested by nickel chelate affinity chromatography method. The antigenicity of recombinant SEO was tested by Western blot. The stimulating effects of recombinant SEO on mouse lymphocytes was tested by MTT. The activity of Caspase 3 and the ap-optosis of DNA ladder were tested. Results The nucleotide sequence of the cloned seo gene was the same as that of reported in GenBank. The soluble recombinant protein GST-SEO was expressed at 25% expression level, and P28-SEO was 22%. The Western blot by GST-SEO-positive sera demonstrated that the recombi-nant SEO had good immunogenicity. The MTY assay of the enterotoxin activity showed that low dose of re-combinant SEO stimulated the proliferation of mice spleen lymphocytes, higher doses will lead to apoptosis. Conclusion Two of the recombinant SEO still have superantigen activity, and can lead to apoptosis of mice splenocytes.

Objective To explore the value of echocardiography in evaluation of pulmonary venous in total anomalous pulmonary venous connection (TAPVC).Methods Fifty-five children with TAPVC were enrolled in the study.The data of echocardiography and CT angiography were retrospectively analyzed and compared with intraoperative findings.Results Totally 55 patients with TAPVC were classified into supra-cardiac type (n=24),cardiac type (n=20),infra-cardiac type (n=7) and mixed type (n=4) according to the sites of drainage of pulmonary venous in echocardiography.In 15 patients with obstruction of pulmonary vertical vein,the sites of obstruction in the supra-cardiac type mostly presented between vertical vein and superior vena cava or innominate vein,and the sites of obstruction in the infra-cardiac presented all between vertical vein and hepatic or portal vein.In 4 patients with pulmonary vein stenosis,3 cases with local pulmonary vein stenosis were all cardiac type,which presented between individual pulmonary vein and common confluence or right atrium;1 patient with diffuse pulmonary vein stenosis was infra-cardiac type.In 9 patients of abnormal individual pulmonary vein,8 cases were not detected by echocardiography,but all were detected by CT angiography.Conclusion Echocardiography is able to make more comprehensive evaluation for the pulmonary venous drainage,obstruction,and proximal stenosis of individual pulmonary vein in TAPVC.CT angiography is superior in evaluation of abnormalities of connection and amount of individual pulmonary vein,and imaging of distal pulmonary vein.

Objective To investigate the genetic basis of patients with intellectual disability,and to assess the application of single nucleotide polymorphisms (SNP)-array in the molecular diagnosis of intellectual disability.Methods Sixty-four patients with intellectual disability who were identified in Maternal and Child Health Hospital of Guangxi Zhuang Autonomous Region from January 2013 to June of 2015 were enrolled.Genomic DNA was extracted from peripheral blood and was analyzed with Illumina Humancyto SNP-12 300K gene array chip.All identified copy number variants (CNVs) were analyzed with references from databases such as ClinVar,DECIPHER,OMIM and DGV(Database of Genomic Variants),as well as comprehensive literature review from PubMed database to determine the pathogenicity of CNVs.Results Sixteen cases of the above 64 patients were found to have CNVs with genomic alterations,including 6 cases microdeletions/microduplications associated with known syndromes,3 cases microdeletions and microduplications with clear clinical relevance (non-syndrome),1 case numerical chromosome aberration,1 case unbalanced translocation and 5 cases CNVs of unknown clinical significance.The detection rate was 25% (16/64 cases).Among these 16 abnormalities,6 cases of them could not be detected by using karyotyping analysis because their sizes were less than 5 Mb,and the smallest detected missing fragment was 0.53 Mb.Conclusion SNP-array gene chip technique with the advantages of higher efficiency,high-resolution and good accuracy,which can be applied to the genetic diagnosis of intellectual disability.

Objective To understand the usage of national medical insurance negotiated drugs(hereinafter referred to as"negotiated drugs")at Tianjin Second People's Hospital and to provide references for optimizing and adjusting the hospital's drug catalog.Methods A retrospective study was conducted on the usage of negotiated drugs from January 1,2018 to December 31,2022 to compare changes in the unit price of drugs,the quantity and amount of sales,the usage frequency(DDDs)and the daily average cost(DDC),etc before and after the negotiation.Results Between 2018 and 2022,the varieties of negotiated drugs used in the hospital increased from the original 5 to 24.Among the 13 drugs analyzed for comparison,the unit prices of 11 drugs had been reduced after negotiation,and 7 drugs were included in the medical insurance and outpatient-specific disease payment directory.The average decrease in DDC was 36.43%,and the average increase in DDDs was 1 770.31%.The implementation of this policy had increased the accessibility of medication for patients and significantly increased sales quantity.Conclusion The quantity of sales of negotiated drugs significantly increased through reducing the unit price of drugs and including them in the scope of medical insurance payment,etc.These increase the pharmacoeconomic viability of negotiated drugs,effectively reduce the burden on patients,promote rational drug use in hospitals,and improve the access and efficiency of drugs.

Objective:To investigate whether heat shock protein 90 (HSP90) participates in the necroptosis of C57BL/6 mouse neurons and spatial memory impairment induced by Aluminum maltol [Al (mal) 3] through RIP1/RIP3/MLKL pathway. Methods:In March 2022, Thirty-two C57 mice were randomly divided into control group, Low dose group, a medium dose group, and a high-dose group, with 8 mice in each group, and injected intraperitoneally with physiological saline, 20, 40, and 80, respectivelyμmol/kgAl (mal) 3 was administered, it was injected 5 days a week and discontinued 2 days for 60 days. Morris water maze test was used to test the spatial learning and memory ability of mice. Nissl staining was used to observe the pathological changes of brain tissue. The protein expression levels of RIP1, RIP3, MLKL and HSP90 in hippocampus were determined by Western blotting. Results:In the water maze experiment, compared with the control group, the number of mice crossing the platform decreased in each dose group, the difference was statistically significant ( H=9.50, P=0.023), and the number of mice crossing the platform was statistically significant among each dose group ( P <0.05). Compared with the control group, the number of hippocampal nerve cells in each dose group decreased, the arrangement was disordered, and the Nissellite bodies decreased. Western blotting results showed that compared with the control group, the expression level of RIP1 protein in the hippocampus of mice in high-dose group was higher, and the difference was statistically significant ( P <0.05). The expression levels of RIP3, MLKL and HSP90 in hippocampal tissue of mice in medium and high dose groups were increased, and the differences were statistically significant ( P<0.05). After siRNA intervention decreased the expression of HSP90 protein, the expressions of HSP90, RIP1, RIP3 and MLKL in Al (mal) 3 groups were increased, and the differences were statistically significant ( P<0.05) . Conclusion:Through RIP1/RIP3/MLKL pathway, HSP90 is involved in neuronal programmed necrosis and spatial memory impairment induced by maltol aluminum in C57 mice.

Objective:To investigate whether heat shock protein 90 (HSP90) participates in the necroptosis of C57BL/6 mouse neurons and spatial memory impairment induced by Aluminum maltol [Al (mal) 3] through RIP1/RIP3/MLKL pathway. Methods:In March 2022, Thirty-two C57 mice were randomly divided into control group, Low dose group, a medium dose group, and a high-dose group, with 8 mice in each group, and injected intraperitoneally with physiological saline, 20, 40, and 80, respectivelyμmol/kgAl (mal) 3 was administered, it was injected 5 days a week and discontinued 2 days for 60 days. Morris water maze test was used to test the spatial learning and memory ability of mice. Nissl staining was used to observe the pathological changes of brain tissue. The protein expression levels of RIP1, RIP3, MLKL and HSP90 in hippocampus were determined by Western blotting. Results:In the water maze experiment, compared with the control group, the number of mice crossing the platform decreased in each dose group, the difference was statistically significant ( H=9.50, P=0.023), and the number of mice crossing the platform was statistically significant among each dose group ( P <0.05). Compared with the control group, the number of hippocampal nerve cells in each dose group decreased, the arrangement was disordered, and the Nissellite bodies decreased. Western blotting results showed that compared with the control group, the expression level of RIP1 protein in the hippocampus of mice in high-dose group was higher, and the difference was statistically significant ( P <0.05). The expression levels of RIP3, MLKL and HSP90 in hippocampal tissue of mice in medium and high dose groups were increased, and the differences were statistically significant ( P<0.05). After siRNA intervention decreased the expression of HSP90 protein, the expressions of HSP90, RIP1, RIP3 and MLKL in Al (mal) 3 groups were increased, and the differences were statistically significant ( P<0.05) . Conclusion:Through RIP1/RIP3/MLKL pathway, HSP90 is involved in neuronal programmed necrosis and spatial memory impairment induced by maltol aluminum in C57 mice.