Objective In order to investigate the roles of hIGF\|1 in treatment of diabetes mellitus and diabetic syndromes,the gene of human insulin like growth factor type Ⅰ(IGF\|1) was cloned and constructed into eukaryotic expression vector,then the expression and activity were determined. Methods Total cellular RNA of human fetal liver was abstracted and the RT\|PCR amplification of the cDNA fragment was performed.The fragment was cloned into pUCM\|T vector and sequenced.The eukaryiotic expression vector was recombined and transfected into fibroblast cell line,COS\|7.The expression of hIGF\|1 was examined by in situ hybridization and immunohistochemistry.The effect of hIGF\|1 on cultured islet cells was observed by glucose\|stimulated insulin release assay. Results The cDNA fragment of 710bp with additional Kozak sequence was amplified by RT\|PCR.Eukaryiotic expression vector pCI\|neo/hIGF\|1 was constructed and IGF\|1 gene expressed in COS\|7.The biological activity of hIGF\|1 was proved by increasing inslin secretion from islet cells.Conclusion\ The newly constructed vector,pCI\|neo/hIGF\|1 could be transfected into COS7 cells and its expressed product showed to have the biology activity of hIGF\|1.\;[

Objective To understand the genetic characteristics of Enterovirus 71 ( EVT1 ) circu-lating strains of Guangdong province in 2008. Methods We isolated an EV71 strain from the fatal case of the hand, foot and mouth disease during an epidemic of Guangdong in 2008. Its complete genome was se-quenced and analyzed comparatively. Results The results showed that the full length of EV71 GDFS-3 ge-nome( not including poly A tail ) is 7405 bp. No insertion or deletion is detected in the coding region. There are several insertions and deletions in 5'and 3'UTR. Phylogenetic analysis of GDFS-3 and reference strains showed GDFS-3 strain shares the highest nueleotide homology with TW984 strain(96.0% ) but low homology with SIN5865, MS and BrCr( about 81.0% ). GDFS-3 strain also shares the highest amino acid homology with TW984 strain(99.0% ). It clustered with reference strains of CA subgenotype in the phylogenetie tree. The nucleotide identity with CA reference strains is 91.0% -95.0%. Conclusion The phylogenetic analysis based on the entire genome demonstrates that GDFS-3 strain has the nearest genetic relationship with TW984 strains ( isolated in 2004). GDFS-3 may belong to the same subgenogroup ( CA ) with Taiwan predominant strains. Otherwise,Some mutations in 5'UTR of EV71 may play the very important role in heightened viru-lence.

Objective To analyse the changes of metabolic parameters and condition of metabolic syndrome(MS)in females with obstructive sleep apnea-hypopnea syndrome(OSAHS),and explore the interaction between OSAHS and MS.Methods Sixty females undergoing polysomnography were included,and were divided into simple snorer group(n=19),mild OSAHS group(n=21)and moderate to severe OSAHS group(n=20).Epworth sleepiness scale(ESS)scores and metabolic parameters were compared among groups.Forty-one patients with OSAHS were subdivided into group with MS(MS group,n=15)and group without MS(non-MS group,n=26).ESS scores,apnea hypopnea index(AHI),oxygen desaturation index(ODI)and lowest pulse oxygen saturation(LSpO_2)were compared between MS group and non-MS group.Results There was no significant difference in age,body mass index(BMI),total cholesterol(TC),triglyceride(TG),low density cholesterol(LDL),high density cholesterol(HDL),apolipoprotein A-I(apoA-I)and apolipoprotein B(apoB) among simple snorer group,mild OSAHS group and moderate to severe OSAHS group(P>0.05).Correlation analysis revealed ESS was positively related to AHI(r=0.327,P=0.011).ESS scores and proportions of hypertension,fasting plasma glucose(FPG)and MS were significantly higher in moderate to severe OSAHS group than those in simple snorer group(P<0.05).BMI,FPG,TG,proportion of hypertension,ODI and ESS score in MS group were significantly higher than those in non-MS group(P<0.05),while there was no significant difference in AHI and LspO_2 between these two groups(P>0.05). Conclusion ESS score may reflect the severity of OSAHS in females.Females with OSAHS matched for age and BMI have no difference in blood fat.With the increase of severity of OSAHS,glycometabolism can be impaired,the prevalences of hypertension and MS increase.MS can exacerbate the severity of OSAHS,indicating that MS and OSAHS interact with each other.

Objective To investigate the effect of Desert Hedgehog on direct differentiation of neural progenitor cells(NPCs) cultured from embryonic mesencephalon in the rats.Methods We infected DHH into COS7,NIH/3T3 and 9L cells,and detected the expression of DHH in the cells with immunofluorescence,real-time PCR and Western blot.All of the three cells were co-cultured with NPCs isolated from ventral mesencephalon in embryonic SD rats(E13.5) respectively.Immunoreactivities of tyrosine hydroxylase(TH) was detected by immunocytochemistry after 10 days.Results The expression of DHH in COS7,NIH/3T3 and 9L cells was remarkably detected,but few TH-positive cells were found in the three co-cultral systems at the 10th day.Conclusion The protein derived from DHH itself does not show any inductive effect on the differentiation of NPCs to the dopaminergic neurons in vitro.

Objective To study the role of phosphorylation at Serine 129 in regulating the neurotoxicity of ?-synuclein. Methods Wild type and phosphomimic mutant ?-synuclein genes were over-expressed in mouse dopaminergic cells MN9D using retrovirus. The cell viability was examined using CCK-8 assay and cell morphology was observed by immunofluorescence microscopy. Results The result of real time PCR showed that WT/?-SYN and S129D/?-SYN genes were overexpressed in MN9D as compared to uninfected MN9D and vector control group(P

Objective To investigate the incidence of nutritional risk for stroke patients with diabetes mellitus. Methods All the stroke patients with diabetes mellitus in a neurologic department were investigated with Nutritional Risk Screening (NRS2002). Results The incidences of undernutrition and nutritional risk were 11.3% (42/372) and 35.8% (133/372), respectively. The incidence of nutritional risk was more in the patients aged over 60 years than in the patients below 60 years (P=0.001). The incidence of both undernutrition and nutritional risk was more in the patients with stroke relapsing than those of the initial stroke (P<0.001). Conclusion The stroke patients with diabetes mellitus are in the risk of undernutrition and nutritional risk, especially those over the age of 60 years and relapsed stroke.

Objective:To analyze the laboratory test results of two outbreaks of neonatal enterovirus infections in Guangdong Province in 2019 and the genetic characteristics of Echo11, aiming to provide reference for the prevention and control of neonatal enterovirus infections.Methods:The pathogenic specimens of neonatal cases suspected of enterovirus infection were collected. Fluorescence quantitative PCR and sequencing were used for enterovirus typing and identification, and virus isolation was carried out for positive specimens.The complete sequences of VP1 of Echo11 were amplified and sequenced and the phylogenetic analysis was performed using the bioinformatics software such as Danstar6, Bioedit7.09 and MEGA6.06.Results:A total of 93 specimens from 36 neonatal cases were collected. After identification, 55 specimens from 24 cases were positive for enterovirus, of which 23 cases were positive for Echo11 and one case was positive for Coxsackievirus B4(CVB4). A total of 29 enterovirus strains were isolated from the specimens of 19 cases, of which 28 were Echo11 from 18 cases, and one was CVB4. Phylogenetic analysis revealed that the nucleotide homology between the 18 strains of Echo11 in this study was 98.2%-100.0%, and the nucleotide homology between the Echo11 strains causing the two neonatal infections was 99.7%-100.0% and 99.8%-100.0%, respectively. Echo11 could be divided into six genotypes as A, B, C, D, E and F, in which genotype A and genotype C were further divided into A1-5 and C1-4, and genotype D could be divided into D1-5. The 18 strains of Echo11 in this study were all subtype D5.Conclusions:In 2019, two outbreaks of neonatal infections in medical institutions in Guangdong Province were caused by Echo11, which belonged to the genotype D5.

The theory of "equal stress on bones and muscles" emphasizes that "the tendons bind to the bones, the bones are stretched, the bones are connected, and the bones are fractured. The relationship between bone and soft tissues are important, which is the law of Traditional Chinese Medicine in the treatment of orthopedic diseases. For patients with lumbar disc herniation, the percutaneous intervertebral foraminal technology remodels the disordered internal biological balance of the spine under pathological conditions. Among them, two common clinical minimally invasive approaches under endoscopy are paid attention to soft tissue protection, and active and appropriate functional exercises after surgery, which have become a typical manifestation of the theory of "equal stress on bones and muscles" in modern spinal orthopedic surgery.

Objective: To investigate the effect of cannabinoid receptor-2 (CB2) agonist AM1241 on the mRNA and protein expression of platelet-derived growth factor (PDGF) and collagen-III (Col-III) in the liver tissue of mice with experimental liver fibrosis induced by carbon tetrachloride (CCl₄). Methods: Totally 38 8-week-old male C57BL/6J mice were randomly divided into control group, model group, 3 mg/kg CB2 receptor agonist (AM1241) group, and 9 mg/kg AM1241 group. All mice, except for the control group, were treated with 30% CCl₄ (three times a week, 5 ml/kg body weight, 16 weeks) to establish a liver fibrosis model. Meanwhile, 3 and 9 mg/kg AM1421 was intraperitoneally injected for daily intervention, respectively. The dosage was adjusted according to actual body weight. The same solvent was given in the control group. The serum level of aspartate aminotransferase (AST) was measured by serum enzyme digestion. The liver inflammation and fibrosis were observed by HE staining of tissue slices. The mRNA and protein expression of PDGF and Col-III in hepatic tissue was determined by real-time PCR and immunohistochemistry. Results: Compared with the control group, the mice in model group showed severe liver fibrosis, significantly elevated serum AST level (742 ± 300.8 U/L vs 118.1 ± 31.1 U/L, P < 0.05), and significantly increased mRNA and protein expression of PDGF and Col-III in liver tissue (P < 0.05). Compared with the model group, the mice in 3 mg/kg AM1241 group and 9 mg/kg AM1241 group had less severe liver fibrosis, and significantly reduced serum AST levels (116.6 ± 13.68 U/L vs 742 ± 300.8 U/L, P < 0.05; 113.8 ± 16.01 U/L vs 742 ± 300.8 U/L, P < 0.05) and mRNA and protein expression of PDGF and Col-III in liver tissue (P < 0.05). Conclusion CB2 receptor agonist AM1241 can inhibit the mRNA and protein expression of PDGF in the liver tissue of mice with hepatic fibrosis, and reduce extracellular matrix synthesis.

Objective : To explore from the perspective of microorganisms the changes in plaque microbial community of children with severe early childhood caries (S-ECC) before and 3 months after dental treatment. Meanwhile to show the effect of treatment on the maintenance of long- term caries-free state. Methods: S-ECC children completed dental treatment under general anesthesia. We collected plaque from caries-free dental surfaces before treatment (caries, C) and at the postoperative follow-up review time points of 7 days (C-7D), 1 month (C-1 M), and 3 months (C-3 M). We included caries-free children (caries free, CF) as the control group to analyze the dynamic modification process of the plaque microbial community in the short-term pre- and postdental treatment. Results: Species clustering analysis showed that the compositions of the microbial communities of the S-ECC and CF groups were highly similar. The α diversity index was not statistically significant (P>0.05). From the analysis of the relative abundance, Leptotrichia spp. and Aggregatibacter spp. decreased after treatment compared with before treatment (P<0.05). Streptococcus sanguinis in the C-7D group increased compared with that in the C group and gradually decreased within 3 months. Veillonella spp., Actinomyces spp., Allprevotella spp., Capnocytophaga spp., and Streptococcus mutans differed between the C and CF groups (P<0.05), Streptococcus mutans did not differ significantly between the C-7D and C-1 M groups and the CF group after treatment, while C-3 M showed an increase compared with the CF group (P<0.01). Conclusion The rapid change in the structure of the flora of children with S-ECC after treatment. The plaque microbial community structure in a caries-free state gradually starts to be established 1-3 months after treatment. There is a "core microbiota" in the oral plaque community that jointly maintains microecological stability. Veillonella spp., Allprevotella spp. and Streptococcus mutans have potential as possible microbial markers.