Clinical, skeletal muscle pathological and genetic characteristics of fatal infantile hypertonic myofibrillar myopathy / 中华实用儿科临床杂志

Objective:To investigate the clinical, skeletal muscle pathological, and genetic characteristics of fatal infantile hypertonic myofibrillar myopathy (FIHMM).Methods:The clinical manifestations, laboratory assessments data and gene sequencing results of 10 patients diagnosed with FIHMM in Shenzhen Children′s Hospital from February 2017 to April 2021 were retrospectively analyzed.Magnetic resonance imaging (MRI) of both musculoskeletal system and the brain, and electromyogram (EMG) were performed in 3 cases, while muscle biopsy was performed in 2 cases.Results:Among these 10 cases, 1 case was from Northeast China and 1 case from East China, while the rest 8 cases were from South China.Eight of the 10 patients were male, and the other 2 cases were female.They were all born normal and not related to each other.The age of onset varied from 2 to 12 months.The main clinical manifestations for all the patients were progressive rigidity of the rectus abdominis (8 cases), neck muscles (7 cases), rectus abdominis (2 cases) and intercostal muscles (1 case), resulting in respiratory failure.Mildly to moderately elevated serum creatine kinase level was detected (436-5 804 IU/L) (reference range: 24-229 IU/L). Complex repetitive discharges can be seen in the EMG, without any myotonic potential.Muscle fiber degeneration, necrosis, and vacuolar degeneration were noted in the histopathological examination of the vastus lateralis and rectus abdominis.An abnormal red granular deposit was observed in a portion of the field of the modified Gomory Trichrome staining.Immunohistochemistry showed substantial deposition of desmin.Under the electron microscopy, the sarcomere structure of the muscle fibers was seriously disordered, with the destruction of Z-bands and the presence of granular deposits.The whole-exome sequencing identified the same homozygous variation c. 3G>A, p.Met1? of CRYAB gene in all the patients, but heterozygous variation in their parents. Conclusions:Axial muscles involvement, such as rectus abdominis rigidity, is the main clinical characteristic of FIHMM.c.3G>A, p.Met1? mutation in the CRYAB gene is a hotspot mutation in Chinese children.

Expression and significance of growth factor receptor binding protein 2 -associated binding protein -1 and glioma-associated oncogene homologue -1 in pediatric medulloblastoma / 中华实用儿科临床杂志

Objective To explore the expressions of growth factor receptor binding protein 2 -associated bin-ding protein -1 (Gab -1 )and glioma -associated oncogene homologue -1 (Gli -1 )in pediatric medulloblastoma,and to analyze their correlation between clinical and pathological characteristics and prognosis in pediatric medulloblastoma. Methods Elivision immunohistochemistry was used to detect the expressions of Gab -1 and Gli -1 protein in tissue microarray of 40 paraffin embedded pediatric medulloblastoma specimens.Chi -square test or Fisher exact test was used to analyze the correlation between Gab -1 and Gli -1 protein expressions with gender,age,tumor location and pathological subtypes.Follow -up data were handled by using Kaplan -Meier survival analysis and Cox regression anal-ysis.Results Positive expression ratios of Gab -1 and Gli -1 protein in 40 pediatric medulloblastoma were 35.0%and 55.0%,respectively.The positive expression rate of Gab -1 in medulloblastoma tissues had no statistical signifi-cance between different genders[male:30.4%(7 /23 cases)vs.female:41 .2%(7 /17 cases)],age[0.05).There were statistical differences of positive expression rate of Gli -1 protein in different age groups[0.05).Kaplan -Meier survival analysis showed that the age,the expressions of Gab -1 and Gli -1 protein were correlated with prognosis of pediatric medulloblastoma(all P <0.05).Cox regression indicated that the age,pathological subtypes and the expression of Gli -1 protein were independent prognostic indicators in pediatric medulloblastoma(all P <0.05).Conclusion Expression of Gab -1 and Gli -1 protein is significantly correlated with the prognosis of medulloblastoma,and the positive expression is a marker of unfavorable prognosis.

New phenotype caused by a NR5A1 heterozygous mutation—— 46, XX disorder of sex development / 中华内分泌代谢杂志

Objective To summarize the clinical manifestations of 46, XX ovotesticular disorder of sex development (DSD) caused by a NR5A1 heterozygous mutation. Methods The first case of 46,XX ovotesticular DSD was caused by a NR5A1 heterozygous mutation in China and was reported with a review of 11 similar cases in the literatures since July 2016. Results A 5. 6-year-old child raised as female was born with ambiguous genitalia. The left gonad was palpable in the inguinal region while the right one was located in abdomen. Gonadal histology showed both ovotestis. Vaginoscopy revealed a short, blind-ending vagina. No uterine was detected by laparoscopy. Repeated karyotype results were 46, XX with SRY gene negative. A heterozygous de novo mutation ( p. Arg92Trp) in the accessory DNA-binding region of NR5A1 gene was found in that child. Conclusions We reported for the first time in China a new phenotype caused by a NR5A1 heterozygous mutation-46,XX ovotesticular DSD. According to the review of literatures, such mutation seemed with incomplete penetrance. It could cause both 46, XX DSD and 46, XY DSD with varied manifestations. The possible underlying mechanism might relate to the impairment of the binding between the mutant protein and target DNA which might lead to a decreased inhibition of the male developmental pathway through downregulation of female antitestis genes.

New phenotype caused by a NR5A1 heterozygous mutation—— 46, XX disorder of sex development / 中华内分泌代谢杂志

Objective To summarize the clinical manifestations of 46, XX ovotesticular disorder of sex development (DSD) caused by a NR5A1 heterozygous mutation. Methods The first case of 46,XX ovotesticular DSD was caused by a NR5A1 heterozygous mutation in China and was reported with a review of 11 similar cases in the literatures since July 2016. Results A 5. 6-year-old child raised as female was born with ambiguous genitalia. The left gonad was palpable in the inguinal region while the right one was located in abdomen. Gonadal histology showed both ovotestis. Vaginoscopy revealed a short, blind-ending vagina. No uterine was detected by laparoscopy. Repeated karyotype results were 46, XX with SRY gene negative. A heterozygous de novo mutation ( p. Arg92Trp) in the accessory DNA-binding region of NR5A1 gene was found in that child. Conclusions We reported for the first time in China a new phenotype caused by a NR5A1 heterozygous mutation-46,XX ovotesticular DSD. According to the review of literatures, such mutation seemed with incomplete penetrance. It could cause both 46, XX DSD and 46, XY DSD with varied manifestations. The possible underlying mechanism might relate to the impairment of the binding between the mutant protein and target DNA which might lead to a decreased inhibition of the male developmental pathway through downregulation of female antitestis genes.

Effect of Risk Classifying Method on Prophylactic Application of Antibiotics in TypeⅠOrthopedic and Paren-chyma Incision / 中国药房

OBJECTIVE:To standardize periooperative prophylactic application of antibiotics. METHODS:According to the characteristics of orthopaedic and parenchyma surgery,classifying evaluation table of typeⅠincision infection risk was designed sci-entifically and rationally. The individual application of antibiotics in surgery patients had been achieved through infection risk evalua-tion. High risk typeⅠincision patients used antibiotics rationally and low risk patients seldom used or didn’t use at all. RESULTS:Through using infection risks classifying table,the rate of antibiotics prophylactic application in typeⅠincision drops from 74.10%to 28.68%,and and the per capita duration of antibiotics prophylactic application shortened from 4.23 d to 2.21 d. The postopera-tive infection rate remained the same. CONCLUSIONS:Through infection risk classifying evaluation,individual application of anti-biotics can be achieved in surgery patients,so as to promote rational use of antibiotics for prophylactic use,reduce antibiotics dos-age and antibiotics abuse under the condition of controllable surgery infection.

Epidemiological survey of Campylobacter infection associated with acute bacterial infectious children diarrhea in Shanghai / 中华检验医学杂志

Objective To assess the prevalence of Campylobacter in the children with acute bacterial diarrhea in Shanghai.Methods Epidemiological survey.Totally 6 641 children with acute bacterial infectious diarrhea from outpatients and inpatients in Children′s Hospital of Fudan University were submitted to the investigation during January 2011 to December 2012.The Campylobacter was isolated from stool samples collected from subjects in micro aerobic environment and identified by both multi-PCR and matrix-assisted laser desorption ionization-time-of-flight mass spectrometry ( MALDI-TOF MS ) .Antimicrobial susceptibility tests were assayed by disk-diffusion method according to EUCAST standard.The isolates molecular typing was done by PFGE.SPSS16.0 was used to analyze the results.Results A total number of 6 641 subjects were enrolled, among them, 305 patients were infected with Campylobacter.The prevalence rate was 4.6%(305/6 641).Among the infected patients, 240 patients were infected with Campylobacter jejuni and 65 patients were infected with Campylobacter coli, the infectious rates of these two pathogenic bacteria were 3.6%and 1.0%, respectively.The peak infectious rate in patients older than 1 year of age was 6.2%(209/3 385) which was higher than that in children under 1 year of age (2.9%, 96/3 256),χ2 =35.98,P<0.001.The infectious rate in winter and spring (6.8%, 138/2 040) was higher than that in the other seasons ( 3.6%, 167/4 601 ) ,χ2 =28.59, P <0.001.Antimicrobial susceptibility test results showed that 91.5%( 279/305 ) isolates were resistant to ciprofloxacin and 11.8%( 36/305 ) isolates were resistant to erythromycin.A total of 9 genotypes of Campylobacter were found by PFGE cluster analysis.The similarity were ranged from 65.1%-100.0%for type A, 67.6%-100.0%for type B, 61.7%-100.0%for type C, 59.0%-100.0%for type D, 71.4%for type F, 80.0%for type H, 54.4%-90.9%for type I, and only one strain was classified as type E and G.Conclusions Campylobacter is a major pathogenic bacteria associated with acute bacterial infectious diarrhea in children especially in children older than 1 year of age in Shanghai.The prevalent pattern of this pathogen was sporadic and the sharp peak was in winter and spring.The isolates are highly resistant to ciprofloxacin but still sensitive to erythromycin.

Distribution and evolvement of resistance profiles of the pathogens in children diagnosed with urinary tract infection in Children's Hospital of Fudan University in the recent 10 years / 中华传染病杂志

Objective To investigate the distribution and evolvement of resistance profiles of the pathogens isolated from children diagnosed with urinary tract infection (UTI).Methods The pediatric patients who were diagnosed with UTI in Children's Hospital of Fudan University from January 2001 to December 2010 were enrolled in this retrospective study.The clean catch midstream urine samples were collected and the pathogens were isolated by culture.Bacterial susceptibility test was performed by following a protocol of the standardized Kirby Bauer (KB) method.The enumeration data were analyzed by chi square test and the tendency of drug resistant rate was analyzed by curvilinear regression.Results A total of 10 686 pathogen strains were isolated during the past 10years.The major pathogen was Escherichia coli which counted for 38％ -56％ of the isolates.However,the proportion of Enterococcus faecalis (0 - 17％,F =34.075,P =0.000) and Enterococcus faecium (3 ％ - 15 ％,F =7.139,P =0.028) increased rapidly.The resistant rates of Escherichia coli to cefazolin,cefuroxime sodium,cefotaxime and ceftazidime were 35.8％- 62.3％(F=215.735,P=0.000),33.1％ -57.0％(F=70.674,P=0.000),19.0％ -56.5％(F=52.355,P=0.000) and 2.1％-23.5％(F=16.807,P=0.003),respectively.The resistant rate of Escherichia coli to ciprofloxacin was 23.4％ - 33.8 ％ (F=0.989,P=0.349),while the resistant rate of Escherichia coli to amikacin and cefoperozone/sulbactam were both less than 7％.There was no Escherichia coli isolate showing resistant to imipenem.The resistant rates of Enterococcus faecium to ampicillin and ciprofloxacin were higher than 60.0％ and 50.0％,respectively,while the resistant rate of Enterococcus faecalis to these two antibiotics decreased from 40.0％ to 9.6 ％ (F =17.497,P =0.009) and 60.0％ to 21.6％ (F=12.826,P=0.009),respectively.The resistant rate of Enterococcus faecium to nitrofurantoin decreased from 32 ％ (in the year of 2002) to 9.4％ (in the year of 2010,F=34.075,P=0.000) and the resistant rate of Enterococcus faecalis decreased from 9.2％ (in the year of 2005) to 1.4％ (in the year of 2010,F=7.139,P=0.028).The resistant rates to erythromycin were both higher than 75.0％ in both Enterococcus in recent 10 years.There was no Enterococcus isolates showing resistant to vancomycin.Conclusions Escherichia coli,Enterococcus faecalis and Enterococcus faecium are the most common pathogens associated with UTI in children.The proportions of Enterococcus faecalis and Enterococcus faecium among the pathogens of UTI are increasing.The antimicrobial activity of ampicillin to Enterococcus faecalis keeps at a high level.However,there are few drugs can be used for treating Enterococcus faecium infection.

Expression of PTEN, p53, Ki-67 and its relationship with tumor histopathological grade and proliferation in astrocytoma / 肿瘤研究与临床

Objective To study the incidence of PTEN, p53 and Ki-67 expression in astrocytoma and show the relationship between PTEN, p53 expression and the proliferation activity. Methods The surgical specimens from 68 brain astrocytoma patients were analysed to detect PTEN, p53 and Ki-67 expression with immunohistochemical method. Results The incidence of PTEN, p53 and Ki-67 expression was 54.4 %,45.6 % and 48.5 % respectively in astrocytoma. With the grade of astrocytoma increasing the levels of PTEN protein decreased, on the other hand the levels of p53, Ki-67 increased. There was a negative correlation between PTEN expression and grade of astrocytoma while there was a positive correlation between p53, Ki-67 expression and grade of astrocytoma by using the Spearman Correlation test to analyse the data. The incidence of Ki-67 positive expression was 24.3 % in 37 cases exhibiting PTEN positive staining, whereas the incidence of Ki-67 positive expression was 77.4 % in 31 cases exhibiting PTEN negative staining. In statistics, there was an inverse correlation between PTEN and Ki-67 expression. Conclusion There is an inverse correlation between histopathological grades of astrocytoma and PTEN expression. A positive correlation is found between p53, Ki-67 expression and histopathological grades of astrocytoma. PTEN can inhibit tumor cell proliferation in astrocytoma.

pcDNAL1 genetic immunization can induce specific cell-mediated immune responses in C57BL/6 mice / 中华医学杂志(英文版)

<p><b>OBJECTIVE</b>To investigate the specific cell-mediated immune efficacy of the an HPV16 prophylactic vaccine.</p><p><b>METHODS</b>C57BL/6 mice were randomly divided into 3 groups: experimental group I (treated with pcDNA L1), control group II (treated with pcDNA3.1) and control group III (treated with PBS buffer). The mice were immunized three times during a three-week interval. Ten to fourteen days after the third inoculation, a footpad swelling test was used to detect delayed-type hypersensitivity (DTH) responses. Antigen-specific splenocyte proliferation assay and quantitation of IFN-gamma cells in splenocytes were performed by FACS assay.</p><p><b>RESULTS</b>In the experimental group, splenocytes actively proliferated after stimulation with HPV16 VLP, and had developed a markedly larger amount of CD8(+) IFN-gamma(+) cells, which is an index for special CTL. Also, the footpad was significantly thickened upon inoculation with HPV16 VLP.</p><p><b>CONCLUSION</b>Naked DNA vaccine of HPV16 L1 can induce specific cell-mediated immune responses in mice, which should be considered for evaluation of HPV16 DNA vaccine feasibility.</p>

Immunological Mornitoring After TIL Treatment of Advanced Human Malignancy / 中国肿瘤生物治疗杂志

Tumor infiltrating lymphocytes(TILs) were isolated by enzymatic digestion and discontinuous gradient centrifugation from 8 human advanced tumors (4 stomach carcinoma, 2 liver cancer, 1 non-small-cell lung carcinoma and 1 colon cancer). These cells were cultured in complete RPMI 1640 medium supplemented with l000U/ml of rhIL2 for 4-6 weeks, till the cell number reach over l09/total, reinfused to the same patients i.v. meanwhile, the patients received 105U of rhIL2 i.m for 5 days. One week before and one month after TIL infusion periphery blood from the patients was collected and the mononuclear cells were isolated. Cytotoxicity against a panel of tumor cell targets by MTT colorometric assay and lymphocyte phenotype by two-color flow cytometry were mornitored. The results showed that there was significant increase in the killing ability to the tested tumor targets to different extent, especially the killing to the target cells which shared the same histological type with the patients tumor. (43 against 1249 lytic units p

The method of directly assessing the immune-protective activity of HPV16L1 antibody / 中国免疫学杂志

Objective:To observe whether the HPV16L1VLP Hela-cell binding-inhibition experiment can assess the immune-protective activities of HPV16L1 antibody in HPV16 prophylactic vaccine or other fields.Methods:C57BL/6 mice were divided into three groups,including pcDNAL1 plasmid,HPV16L1VLP protein,pcDNA3.1 plasmid.Each group of mice was immunized intramuscularly or subcutaneously with certain above antigens,accompanying three times and interval 3 weeks.After 14 days of 3 rd immunization,take blood to make serum for immunochemistry.Results:Hela cell neutralized by the serum IgG of experimental mice can be stained negatively,while the cells of control and non-inoculation groups can be stained positively (brown-yellow color).Conclusion:HPV16LIVLP Hela-cell binding-inhibition assay may be the better way to reflect the neutralizing-protective activities of HPV16L1 antibody in many fields including HPV16 prophylactic vaccine.