Phospholipase A2(PLA2) is an enzyme family which widely distributed in nature. It is related to many important function in physiology and pathology. Among the enzymes, a small molecule PLA2 (platelet PLA2) is closely related to tumor. The study of platelet PLA2 and tumor is reviewed.

Objective To screen polymorphisms and mutations in the promoter region of WD gene.Methods DNA from peripheral blood was obtained from 71 subjects of 36 family (48 WD patients,23 patients first-degree relatives) and 20 healthy people from Feb.2001 to Feb.2004.DNA sequence of the genes was analyzed by PCR amplification and direct genomic sequencing.Results There were three polymorphisms at positions-190,-78,+260(transcription start site as +1) of the promoter region of WD gene.Normal controls,WD patients and patients’ first-degree relatives all showed the polymorphisms;three of 48 WD patients presented C→T base substitution mutations at the same position -183:two were homozygous mutation,while the other was heterozygous.Normal control subjects and patients' relatives didn’t show this kind of mutation.Conclusion It suggests that the mutation of the promoter region is one of WD pathogenesis.

Objective To study the effects and mechanism of exercise on learning-memory of space and in aged rats after cerebral ischemia-reperfusion. Methods 20 SD rats at the age of 20 months with cerebral ischemia-reperfusion were randomly divided into 2 groups. The experimental group (n=10) received regularly exercise, while the control group (n=10) exercised freely. Morris water maze was tested 1 week after exercise, and the brain-derived neurotrophic factor (BDNF) mRNA expression on the ischemic side of the hippocampus were detected.Results The latency in the 2nd round decreased (P<0.05) and the path line on the platform increased (P<0.05) in the experimental group in Morris water maze test (P<0.05), while the BDNF mRNA increased (P<0.01), compared with control group. Conclusion Regular exercise can improve space learning-memory ability of cerebral ischemia-reperfusion aged rats, which may associate with increased BDNF mRNA expression in hippocampus.

Objective 　To study the sequence and structure of intron 8 in WD gene in order to further understand the relationship between intron 8 and WD. Methods　We utilized polymerase chain reaction (PCR) to the amplification of exon 8-intron 8-exon 9 which were then sequenced by a dideoxy chain termination methon in 10 normal controls and 32 members of 11 families(20 WD patients and 10 of their relative). The results were analyzed by the computer. Results　The sequence of intron 8 was 703 bp with the G　+　C content of 42.7%. There were one short tandom repeats, 7 direct and inverted repeats in it. An open reading frame coded with 82aa was found at 323 base pairs of downstream of a TATAbox. There were two DNA polymorphisms at 408 and 487 nucleotides. The sequence analysis showed that the 5end has the sequence of 5-GTAAC, 3end has the sequence of CCTAG-3, and branchpoint of 5-TTTCGA-3.Conclusions　The sequences and structures of intron 8 in WD familiess members are not different from normal controls. Our data suggest that the WD gene intron 8 might not play an important role in the pathogenesis of WD.

OBJECTIVETo study the role of integrin-linked kinase (ILK) on the proliferation and differentiation of human fibroblast in hypertrophic scar and its effect on the scar formation.

METHODSThe human scar fibroblasts were isolated and cultured in vitro. The cells were divided into 4 groups. (1) control group: only contains DMEM; (2) jetPRIME group: DMEM with 200 microl jetPRIME buffer and 4 microl jetPRIME; (3) ILK siRNA group: DMEM and ILK siRNA; (4) ILK cDNA group: DMEM and ILK cDNA. The cell proliferation was detected by XTT assay and the mRNA and protein expressions of ILK and alpha-SMA were detected by Real-time qPCR and Western blot.

RESULTS(1) XTT results showed that the cellular proliferation level after 48 h in four groups were 0.820 +/- 0.065, 0.873 +/- 0.041, 0.554 +/- 0.013 and 1.296 +/- 0.094, respectively. The cellular proliferation curve showed that the cellular proliferation level was very flat in ILK siRNA group while the cellular proliferation level gradually increased from 12 h. 48 h after transfection, the cellular proliferation level in ILK siRNA group was significant lower than those in other groups (P value were 0.021, 0.034, 0), while the cellular proliferation level in ILK cDNA group was the highest among all 4 groups (P value were 0.017, 0.009, 0). (2) The Real-time qPCR showed that the expressions of ILK mRNA and alpha-SMA mRNA were 0.693 +/- 0.412 and 0.422 +/- 0.037 in control group, were 0.621 +/- 0.183 and 0.388 +/- 0.005 in jetPRIME group, were 0.052 +/- 0.019 and 0.073 +/- 0.023 in ILK siRNA group, were 240.193 +/- 35.170 and 138.056 +/- 24.060 in ILK cDNA group. The expressions of ILK mRNA and alpha-SMA mRNA in ILK siRNA group were significantly lower than those in other three groups (P < 0.05). And the expressions of ILK mRNA and alpha-SMA mRNA in ILK cDNA group were significantly higher than those in other three groups (P < 0.05). (3) The Western blot also showed that the expression of ILK and alpha-SMA proteins were decreased in ILK siRNA group and increased in ILK cDNA group.

CONCLUSIONILK may promote the proliferation and differentiation of human scar fibroblast. It may play an important role in scar formation and contracture.

Actins ; metabolism ; Adolescent ; Adult ; Cell Differentiation ; drug effects ; Cell Proliferation ; drug effects ; Cells, Cultured ; Cicatrix, Hypertrophic ; metabolism ; Female ; Fibroblasts ; cytology ; drug effects ; metabolism ; Humans ; Male ; Middle Aged ; Protein-Serine-Threonine Kinases ; pharmacology ; RNA, Messenger ; genetics ; Transfection ; Young Adult

Objective To investigate the infection and epidemiological characteristics of group A rotavirus (RV-A)and adenovirus in children with diarrhea in Guangzhou. Methods The colloidal gold technique was used to detect RV.A and adenovirus antigen in 2,171 stool samples from children with diarrhea in Guangzhou Women and Children′s Medical Center from January to December 2015,and the data were statistically analyzed. Results Among the 2,171 patients,the positive rate of RV-A infection was 17.96%and that of adenovirus infection 8.66%, and the co-infection rate of both virus was 3.45%. The positive rates between different genders were not significantly different(P > 0.05);the infectious time peak of RV-A was January(40.78%),followed by December(39.24%) and February(32.61%)and that of adenovirus infection was July(15.89%)and May(15.79%). The infectious peak of RV-A and adenovirus was December(7.29%),followed by January(7.01%). The peak age of infection ranged from 1y to 3y. Conclusion RV-A and adenovirus are the main pathogens of children diarrhea ,and the onset of virus infection has obvious seasonal change.

Objective To understand the molecular epidemiology of penicillin resistance Streptococcus pneumonia (PNSP) isolated from children in Guangzhou area to provide the experimental basis for clinical prevention and control of Streptococcus pneumonia infectious diseases.Methods Specific primers were designed according to Genebank,penicillin binding protein(PBP) genes PBP1A,PBP1B,PBP2A,PBP2B,PBP2X,PBP3 were amplified by PCR.The sequencing analysis was performed.The PCR products were digested by Hinf I,and the restriction fragment length polymorphism (RFLP) was analyzed.Results DNA of PNSP was successfully extracted,the PCR results showed that in 50 strains of PNSP,the positive rates of bacterial strains containing PBP1A,PBP1B,PBP2A,PBP2B,PBP2X and PBP3 were 48.9%,64.4%,71.1%,31.1%,40.0% and 31.1% respectively.The sequencing showed that their homologies with known sequences in GenBank were 99%,98%,100%,97%,95% and 100% respectively.Using RFLP in Hinf I showed that PBP1A,PBP1B,PBP2A and PBP3 only had one kind of genotype,PBP2B and PBP2X had two kinds of genotypes,the positive rates were 71.4%,28.6%,66.7% and 33.3% respectively.Conclusion The gene distribution of PNSP strains among children in Guangzhou is dominated by PBP2A,PBP1B and PBP1A,there are two subtypes in PBP2B,PBP2X when digested by Hinf I,in which the predominant subtype >65%.

Objective 　Determination of Wilson disease gene mRNA expression in human fibroblast cell strain (Me32aT22/2L) by reverse transcription-polymerase chain reaction (RT-PCR). Methods　Using lipofection reagent, the plasmid vector carrying the Wilson disease gene (pRc/CMV-WD) was transferred into Me32aT22/2L cultured in serum free complement medium. RT-PCR was used to determine WD mRNA expression in Me32aT22/2L. Results　 Wilson disease gene expression was detected in Me32aT22/2L, while no specific signals were detected in untransfected fibroblast. Conclusions　It demonstrated that Me32aT22/2L strain could express the Wilson disease gene, suggesting that Wilson disease gene transfer might develop a new approach to study Wilson disease.

Objective:To investigate the multilocus sequence typing feature of the virulence-associated genes of Staphylococcus aureus(S. aureus) separated from the clinical specimens of a multi-center cohort children in Guangzhou area. Methods:A total number of 412 Staphylococcus aureus strains isolated from 2 059 non-repeated fecal specimens of children by three groups′ researchers in Guangzhou Women and Children′s Medical Center from August 2018 to November 2018. While collecting specimens, patient clinical information is also properly collected and preserved. After extracting the DNA of the strain, the virulence-associated genes were detected by polymerase chain reaction (PCR), including the staphylococcal enterotoxin (SE) genes ( sea, seb, sec, sed, see) and the Panton-Valentine leucocidin-encoding gene ( pvl).The multi-locus sequence typing (MLST) method was performed to reveal the MLST feature of these genes and the statistical difference were examined by the the χ 2 test. Results:Among the 412 isolates of S. aureus, 256 strains (256/412, 62.1%) contains at least one SE gene. Among the enterotoxin gens, the sec (125/412, 30.3%), seb(98/412, 23.8%)and sea (66/412, 16.0%)genes were the three most prevalent members of SEs. The frequency of pvl gene in Staphylococcus aureus was 18.7%(77/412).Among them, the frequency of Staphylococcus aureus sea gene isolated from patients with gastroenteritis (58/319, 18.2%) was significantly higher than that from the non-gastroenteritis group (8/93, 8.6%)(χ2=4.912, P=0.027). The frequency of Staphylococcus aureus pvl gene isolated from the patients with pneumonia (8/21, 38.1%) was greater than that from the non-pneumonia group (6/47, 12.8%)(χ2=4.252, P=0.039). In addition, the virulence-associated gene of S. aureus was closely related to the specific ST type, 82.4% (28/34) of ST6 carried sea gene, all ST338 and ST59 carried seb gene, 96% (48/50) ST45 carried sec gene, and the pvl gene carrying rate of ST338 was 5/5. Conclusions:The SEA toxin produced by ST6 Staphylococcus aureus may be closely related to the diagnosis of gastroenteritis in children. The frequency of pvl virulence gene in Staphylococcus aureus in children with community-acquired pneumonia was higher than that in the non-pneumonia group, and closely related to the CC59.

OBJECTIVE：To investigate the effects of metformin on malignant phenotype of pancreatic cancer BxPC- 3 cells. METHODS：Using human pancreatic cancer BxPC- 3 cells with natural deletion of Smad4 gene as reaserch objects ，CCK-8 assay and flow cytometry were used to detect the proliferation and apoptosis of BxPC- 3 cells after treated with different doses of metformin（5，10，20 mmol/L）for 24 h. The cell survival rate and apoptosis rate were calculated. Transwell assay was used to test the migration of cells after treated with different doses of metformin （10，20 mmol/L）for 24 h. The number of migrating cells was recorded. qRT-PCR and Western blotting assay were performed to determine mRNA and protein expression of E-cadherin ，Vimentin and RGC- 32 in cells. RESULTS ：Compared with control group and 5 mmol/L metformin group ，survival rate of cells were decreased significantly in 10，20 mmol/L metformin groups ，while apoptosis rate was increased significantly ；the apoptosis rate in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group （P＜0.05）. Compared with control group ， the number of migrating cells was decreased significantly in 10，20 mmol/L metformin groups ，and the 20 mmol/L metformin group was significantly lower than 10 mmol/L metformin group （P＜0.05）. Relative mRNA and protein expression of E-cadherin were increased significantly in 10，20 mmol/L metformin groups ，and relative mRNA expression of E-cadherin in 20 mmol/L metformin group was significantly higher than 10 mmol/L metformin group. Relative mRNA expression of Vimentin in 10 mmol/L metformin group ，relative mRNA and protein expression of Vimentin in 20 mmol/L metformin group ，relative mRNA and protein expression of RGC- 32 in 10，20 mmol/L metformin groups were decreased significantly ；relative mRNA and protein expression of Vimentin as well as mRNA expression of RGC- 32 in 20 mmol/L metformin group were significantly lower than 10 mmol/L metformin group （P＜0.05 or P＜0.01）. CONCLUSIONS ：Metformin can inhibit the proliferation and migration of pancreatic cancer cells through smael-independent pathways in a dose- dependent manner ，and promote their apoptosis ，which is associated with the inhibition epithelial- mesenchymal transition and the expression of RGC- 32 of pancreatic cancer.