To investigate the clinical significance of the expression of CD40L on peripheral blood monocytes and the changes in serum soluble CD40L in patients with acute coronary syndrome, 16 normal controls and fifty-six patients including 24 with SA (8 patients after PTCA), 20 with UA and 12 with AMI entered in this study. The expression of CD40L on monocytes was analyzed by indirect immonofluorescence flow cytometry and serum sCD40L levels were measured by ELISA. The results showed: (1) The expression of CD40L on monocytes in UA and AMI were higher compared with SA and controls ( P 0.05). (2) Patients with UA and AMI had significantly raised serum levels of sCD40L when compared with patients with SA and controls( P

DNA, Viral ; blood ; Female ; Hepatitis B ; immunology ; virology ; Hepatitis B Antibodies ; blood ; Hepatitis B Core Antigens ; immunology ; Hepatitis B Surface Antigens ; immunology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Male ; Polymerase Chain Reaction ; methods

Objective To investigate the 16S rRNA methylase genes and Aminoglycoside modifying enzymes(AMEs)genes in Enterobacter cloacae isolated from the People's Liberation Army 98th Hospital,Huzhou district,Zhejiang province,China.Methods 40 strains of Enterobacter cloacae were isolated from the inpatients between September,2003 and November,2004.5 kinds of 16S rRNA methylase gene (including armA,rmtA,rmtB,rmtC and rmtD)and 9 kinds of AMEs gene[including aac(3)-Ⅰ,aac(3)-Ⅱ,aac(3)-Ⅲ,aac(3)-Ⅳ,aac(6')-Ⅰ b,aac(6')-Ⅱ,ant(3'')-Ⅰ,ant(2'')-Ⅰ and aph(3')-Ⅵ]were analyzed by PCR and verificated by DNA sequencing.Results In 40 strains of Enterobacter cloacae,the positive rates of genes of rmtB,aac(3)-Ⅱ,aac(6')-Ⅰ b,ant(3'')-Ⅰ,ant(2'')-Ⅰ and aph(3')-Ⅵ were 12.5%(5/40),27.5%(11/40),72.5%(29/40),32.5%(13/40),5.0%(2/40)and 5.0%(2/40),respectively.8 kinds of the rest of genes were all tested negative.The total positive rate of AMEs gene was 85.0%(34/40).Among 29 strains of Enterobacter cloacae that the aac(6')-Ⅰ b gene was positive,through PCR and verification by DNA sequencing,7 strains(24.1%)were confirmed to take the aac(6')-Ⅰ b-cr(the GenBank register number:EF375620,EU159121)alone,18 strains(62.1%)were confirmed to take the aac(6')-Ⅰ b-Suzhou(EU085533)alone,3 strains(10.3%)were confirmed to take both aac (6')-Ⅰ b-Suzhou and aac(6')-Ⅰ b-cr while only 1(3.4%)was aac(6')-Ⅰ b(the classical type).Conclusion There was lower positive rate of 16S rRNA methylase gene but very high AMEs genotypes in Enterobacter cloacae isolated from inpatients and the finding of rmtB gene was reported for the first time in the world.At least 5 kinds of AMEs gene existed in Enterobacter cloacae were isolated and they were the new host of both gene of aac(6')-Ⅰ b-cr and aac(6')-Ⅰ b-Suzhou,with aac(6')-Ⅰ b-Suzhou gene was the predominance subtype in aac(6')-Ⅰ b.

OBJECTIVETo study the clinical and molecular characteristics of methicillin-resistant Staphylococcus aureus (MRSA) infection in children.

METHODA total of 37 MRSA strains were isolated from hospitalized patients in Children's Hospital of Fudan University from March 2009 to November 2011. The clinical characteristics were investigated by a cohort study. Furthermore, the mecA, Panton-Valentine leucocidin (PVL) genes were detected by polymerase chain reaction (PCR), and the genotypes of SCCmec were determined by multiplex PCR.

RESULT(1) Among the 37 MRSA isolates, infections with 21 were acquired from hospital (HA-MRSA), and 16 isolates were acquired from community (CA-MRSA). (2) In the study, MRSA frequently caused respiratory tract infection, and most of the strains were isolated from intensive care unit (ICU). (3) CA-MRSA was most frequently associated with skin and soft tissue infections (SSTI), suppurative tonsillitis, even pneumonia and septicemia. HA-MRSA infection was more aggressive, most frequently associated with pneumonia, septicemia, and central nervous system (CNS) infections, such as meningitis. In children with fever caused by HA-MRSA or CA-MRSA infection, HA-MRSA showed a longer duration of fever, for 10.5 days. C-reactive protein (CRP) level caused by HA-MRSA (63.00 mg/L) was higher than CA-MRSA (9.50 mg/L) , and there were statistically significant differences between the groups (t = 2.5670, P < 0.05). However, there were no statistically significant differences between the groups in white blood cell count (WBC) or procalcitonin (PCT) level. (4) Among 37 MRSA isolates, the whole isolates were mecA gene positive (100%). SCCmec genotyping results showed that the most frequent SCCmec types were type III, 17 isolates, the others including type IV 8 isolates, type II1 isolates, nontypable 11 isolates, type I and type V were not found in this group. Therein, among 21 HA-MRSA isolates, SCCmec III was the most common, 15 isolates, type IV 1 isolates, nontypable 5 isolates; among 16 CA-MRSA isolates, SCCmec type IV was the most common, 7 isolates, type III 2 isolates, type II 1 isolate, nontypable 6 isolates. (5) Among the 37 MRSA isolates, 28 were PVL gene positive; and among 21 HA-MRSA isolates, 17 were PVL gene positive; Among 16 CA-MRSA isolates, 11 were PVL gene positive; There were no statistically significant differences between the groups (χ(2) = 0.735, P > 0.05) .

CONCLUSIONCompared with CA-MRSA, HA-MRSA infection was more aggressive, and induced higher C reactive protein; the dominant epidemic strains of CA-MRSA was SCCmec type IV, and HA-MRSA was SCCmec type III; the positive rate of PVL gene was high.

Adolescent ; Anti-Bacterial Agents ; pharmacology ; Bacterial Proteins ; genetics ; Bacterial Toxins ; genetics ; Bacterial Typing Techniques ; Child ; Child, Preschool ; China ; epidemiology ; Cohort Studies ; Community-Acquired Infections ; epidemiology ; microbiology ; Cross Infection ; epidemiology ; microbiology ; DNA, Bacterial ; genetics ; Female ; Genotype ; Humans ; Infant ; Infant, Newborn ; Male ; Methicillin ; pharmacology ; Methicillin Resistance ; genetics ; Methicillin-Resistant Staphylococcus aureus ; classification ; genetics ; isolation & purification ; Penicillin-Binding Proteins ; Staphylococcal Infections ; epidemiology ; microbiology

Using the character of natural aggregation of CHO cells, and an ultrasonic and sedimentation column combined perfusion system to promote cells aggregation and retention into bioreactor,recombinant CHO cell strain MK3-A2 was cultured,which could secrete rhTNK-tPA, by a serum-free perfusion culture system. The culture periods in this two experiments were as long as 77 and 110 days respectively. The cells density reached 2?107 cells /ml. The average volumetric productivity of rhTNK-tPA was 89 mg/L?d, and the highest one was 216mg/L?d.The cells aggregation rate was approximately 90%, and the diameters of most of them were 285～570?m. During the perfusion culture the cells retention rate almost kept in 95% and the viability of cells was more than 85%.Thus, it means that aggregation culture with such perfusion system could be used to scale up produce biopharmaceuticals instead of microcarrier culture system.

After the epidemic of novel Coronavirus Disease 2019(COVID-19), construction of disease prevention and control has become a top priority. As a pioneer in the recovery of global economy and society, Shanghai should play a fundamental role in building a comprehensive system of public health and advanced disease prevention and control in the new era. In this article, we systematically categorize the requirements for the construction of disease prevention and control system in the new era, identify the weakness and challenges during and after the epidemic, and then make suggestions. It is proposed that we should utilize the important window period of the"14th Five-Year Plan", with the"Healthy China"strategy and municipal"20 Tasks for Public Health Construction"as the starting point, to make substantial contribution to the functional orientation, investment of resources, capacity building, operational mechanism and team building, which may provide scientific evidence for the reform and development of disease prevention and control system.

OBJECTIVE: To examine the expression absence of LRRC4 gene in glioblastoma cell lines. METHODS: RT-PCR and Northern blot were used to detect the expression of LRRC4 gene in 6 glioblastomas cells lines. Polymerase chain reaction and DNA sequencing were used to screen the LRRC4 gene mutation, while bioinformation assay was used to search for the reason of LRRC4 gene absence in U251 cell lines. RESULTS: The expression of LRRC4 was absent in 6 malignant glioma cell lines (U251, U87, BT325, SF126, SF767 and M17), which were examined by Northern-blot and RT-PCR assay. All sequencing of PCR products from gDNA of SF126, SF767, and M17 cell lines contained the point mutation at the same position ( LRRC4 geneT977A) (3/5), which was a synonymous mutation. However, PCR products from gDNA of U251 and U87 cell lines (2/5) were not obtained. The expression absence of LRRC4 was ascribed to the loss of homozygosity of 7q32-ter in U251 cell lines. CONCLUSION The expression of LRRC4 gene is absent in glioblastoma cell lines, and it offers the important experiment proof for LRRC4 to act as a new candidate of brain tumor suppressor gene from glioma. The loss of homozygosity of 7q32-ter contributed to the expression absence of LRRC4 in U251 cell lines.
Base Sequence ; Blotting, Northern ; Brain Neoplasms ; genetics ; pathology ; Cell Line, Tumor ; DNA Mutational Analysis ; Gene Expression Regulation, Neoplastic ; Glioblastoma ; genetics ; pathology ; Humans ; Nerve Tissue Proteins ; genetics ; Point Mutation ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVE: To explore the effect of LRRC4, a glioma suppressive gene, on blocking U251 cells in G0/G1 by MAPK signaling pathway. METHODS: LRRC4 was transfected into U251 cells, and at 24 hour of post-transfection, cells were split at a 1:3 dilution, challenged with 500 microg /mL G418 and formed a stable transfected clone pool. RT-PCR, Northern blot and Western blot were used to identify the stable transfectants. ERK, JNK and P38 expression changes were analyzed by Western blot. FACS analysis, Luciferase reporter gene assay and Western blot were used to detect the cell cycle and cyclin D1. RESULTS: LRRC4 down-regulated the expression of phosphorylated ERK2 and up-regulated the expression of total protein JNK2 (a key molecule of MAPK signaling pathway) and phosphorylated c-Jun. LRRC4 decreased the expression of mutation P53, cyclin D1 activation and its expression. U251 cells were blocked in G0/G1 by LRRC4. CONCLUSION LRRC4 can decrease JNK2, up-regulate the phosphoralated c-Jun, down-regulate mutant P53 and cyclin D1, and therefore block U251 cells in G0/G1.
Blotting, Northern ; Blotting, Western ; Cell Line, Tumor ; Cyclin D1 ; metabolism ; Flow Cytometry ; G1 Phase ; genetics ; physiology ; Glioma ; genetics ; metabolism ; pathology ; Humans ; Luciferases ; genetics ; metabolism ; MAP Kinase Signaling System ; genetics ; physiology ; Mitogen-Activated Protein Kinase Kinases ; metabolism ; Nerve Tissue Proteins ; genetics ; metabolism ; physiology ; RNA, Messenger ; biosynthesis ; genetics ; Recombinant Fusion Proteins ; genetics ; metabolism ; Resting Phase, Cell Cycle ; genetics ; physiology ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

The heat shock proteins (HSPs) 70 and HSP 27 expression in patients with non-small cell lung cancer (NSCLC) was studied and the relationship between HSP 70 and HSP 27 with the clinicopathological features of NSCLC was investigated. The expression of HSP 70 and HSP 27 was detected in tumor tissues from 60 patients with NSCLC by S-P immunohistochemistry. The findings were analyzed in combination with the histological types, histopathological differentiation, lymph node metastasis, patients' clinical stages, smoking history and gender. The results showed that of the 60 NSCLC tissue specimens studied, the immunoreactivity of HSP 70 and HSP 27 was detected in 47 (78.3 %) and 43 (71.7 %) specimens, respectively. A positive correlation was found between the overexpression of HSP 70 and HSP 27. The histopathological differentiation, lymph node metastasis, clinical stages and smoking history were correlated to the expression of HSP 70, but not to the expression of HSP 27. No statistical significance was observed in histological types and gender with respect to both HSP 70 and HSP 27 expression. It is suggested that the HSP 70 expression is a powerful and significant prognostic indicator and is related to histopathological differ entiation, lymph node metastasis, patients' clinical stages, smoking history, whereas HSP 27 expression is not.

OBJECTIVETo study the prevalence and distribution of Tourette syndrome (TS).

METHODSNine thousand, seven hundred and forty-two children and juveniles aged 7 - 16 years in Wenzhou were studied, using cluster random sampling method.

RESULTSThe prevalence of TS among school-age children was estimated to be 0.43% (0.74% for males and 0.07% for females). The prevalence of male children and juveniles was higher than that of female children and juveniles (chi(2) = 25.09, P < 0.001, prevalence ratio = 10.95, prevalence ratio 95% CI: 3.38 - 35.46). The highest prevalence of TS was between 9 - 10 years old. The mean age at onset of TS was 7.7 +/- 2.7 years, with 45.2% of them among 6 - 7 year olds. The rate of delayed diagnosis and rates of misdiagnosis and misclassification of the syndromes were 78.6%, 42.9% and 23.8%, respectively.

CONCLUSIONTourette syndrome had been a common disease of children and juveniles in Wenzhou area. The disease was correlated with age and sex, often misdiagnosed and misclassified. Physicians and as well as general publics should be trained to identify the cases.

Adolescent ; Age Factors ; Child ; China ; epidemiology ; Cross-Sectional Studies ; Female ; Humans ; Male ; Prevalence ; Sex Factors ; Tourette Syndrome ; diagnosis ; epidemiology