OBJECTIVETo analyze the changes of patients with type 2 diabetes in different stages in glucagon (GC) and free fatty acid (FFA) in fasting, OGT and L-Arg experiments, and discusses the role of pancreatic alphabeta cells in diabetes pathogenesis by studying the relations among indexes such as glucagon (GC), free fatty acid (FFA) and blood glucose (BG), insulin, insulin homeostasis model (HOMA) and glucose metabolism hormone secretion curve, in order to provide theoretical basis for the treatment of diabetes.

METHODStudy objects were divided into the T2DM group (45 cases), the IGT group (28 cases) and the NGT group (30 cases) for an OGTT experiment and then an L-Arg experiment on the next day. Under the fasting state, their blood glucose (FBG), insulin (F), glucagon (FGC), free fatty acid (FFA) were detected to calculate HOMA-beta, insulin sensitivity index (ISI) and HOMA-IR of different groups. Meanwhile, efforts were made to calculate different time quantum detected in OGTT and L-Arg experiments and area under the curve AUC(BG), AUC(INS) and AUC(GC).

RESULTObvious overall differences were observed in FFA and FGC of the three groups. FGC of each group was negatively correlated with HOMA-beta and ISI. Among all of the 103 study objects, FGC was positively correlated with FBG and HOMA-IR and negatively correlated with HOMA-beta and ISI, with no correlation with FINS; FFA was positively correlated with FBG, HOMA-IR and negatively correlated with FINS, HOMA-beta, ISI. FGC and FFA were positively correlated in the T2DM group and the IGT group, but with no statistical correlation in the NGT group. The sequence of the three study objects was T2DM > IGR > NGT in AUC(GC) in the OGTT experiment and T2DM > IGR > NGT in in AUC(GC) in the L-Arg experiment, with the significant positive correlation between AUC(GC) and AUC(BG) and significant negative correlation with AUC(INS).

CONCLUSIONGlucagon and free fatty acid of T2DM and IGT patients increased, which was positively correlated with blood glucose and HOMA-IR and negatively correlated with INS, HOMA-beta and ISI. The increase in glucagons of T2DM and IGT patients indicated inappropriate secretion of pancreatic alphabeta cells among patients with type 2 diabetes.

Adult ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; Fatty Acids, Nonesterified ; metabolism ; Female ; Glucagon ; blood ; Humans ; Insulin ; secretion ; Islets of Langerhans ; secretion ; Male ; Middle Aged

In the present study, we inserted the core-streptavidin cDNA into downstream of multi-cloning site of plasmid pOPE101-8E5 by DNA gene recombination technology. And then, the variable fragments of heavy and light chain of the scFv-8E5 were replaced by the scFv-C4 variable fragments to construct the expression vector pOPE101-C4∷core-streptavidin. After transformed the vector pOPE101-C4∷core streptavidin into E.coil, the fusion protein C4∷core streptavidin-His-tag can be expressed by inducing with IPTG, and the expression level and activity of the expressed fusion protein analyzed by SDS-PAGE and Western blot. The results show that a scFv-C4∷core-streptavidin fusion protein of 45kDa was obtained, which can bind proteins of 60kDa & 45kDa from the KG1a cells lysate simultaneously. The binding function can be detected by the binding of core-streptavidin and biotin directly.

Objective: To investigate the clinical characteristics of pertussis-associated pneumonia and analyze it's risk factors. Methods: Clinical data were taken from Shenzhen Children's Hospital with Bordetella pertussis infection and confirmed by culture or real-time polymerase chain reaction (PCR) of nasopharyngeal secretion from October 2013 to December 2015. Patients were divided into two groups, those with radiologically confirmed pneumonia in the course of their disease and those with not. Clinical data were retrospectively analyzed and compared. T test, Rank sum test or chi square test were used for comparison between groups. Risk factors were analyzed by unconditional Logistic regression analysis. Results: A total of 501 children hospitalized with Bordetella pertussis infection were included. Among them, 309 patients were diagnosed with pneumonia. The median age was 3 (2, 6) months. Symptoms were paroxysmal cough (n=252, 81.6%), tachypnea (n=69, 22.3%), and cyanosis (n=105, 34.0%). The time from onset of cough to radiologically confirmed pneumonia was between 1 and 66 days with a median of 9 (5.5, 15.0) days. The most common pathogen of coinfection was respiratory syncytial virus (RSV)(20 cases). Macrolides were used in 306 cases for (8.2±3.6) days. All cases showed significant improvement. There were more male children (62.1% (192/309) vs. 50.3% (95/189) , χ²=6.768, P=0.009), and more instances of comorbidities (13.3% (41/309) vs.5.8% (11/189) , χ²=6.957, P=0.008) in the pneumonia group than in the other. The age was younger (3 (2,6) vs.4 (2,6) months, Z=32.91, P=0.000) in pneumonia group than in the other. Male sex, younger age, and underlying disease were independent risk factors for pertussis-associated pneumonia (OR=1.648, 1.486, 2.695, P=0.008, 0.036, 0.005). Conclusions Pneumonia, as a complication of pertussis, is very easy to see in hospitalized children. The duration of hospitalization is extensive. It is more likely to happen in children who are male, young, and having underlying diseases. Pneumonia is easy to occur in the first 2 weeks of the course of disease.

OBJECTIVETo clone and express VP, gene from HBoV, and the expressed VP, protein was as the antigen in order to detect serum from children in Wenling area with lower respiratory tract infections.

METHODSThe VP, gene was recombined with the genome of Baculovirus, which infected the insect cell. The fusion protein with HA tag was applied to confirm the specificity of expressed protein. Furthermore, the recombinant protein was observed using electron microscopy. The 176 serum from children in Wenling area with lower respiratory tract infections was screened using Western blot.

RESULTSThe expressed VP2 protein was more than 60% in total proteins from insect cell, and MWt about 60 x 10(3). The virus-like particle (VLP) was observed using electron microscopy, and size about 20 nm. The 176 serum from children in wenling area with lower respiratory tract infections was screened using Western blot. The HBoV positive rate was 2.28% (4/176).

CONCLUSIONThe VP2 protein from human bocavirus was expressed in insect cell successfully. Through HA tag the VP2 protein was specific, and then the assay using SDS-PAGE with Western blot could detect and screen the antibody in serum from children with lower respiratory tract infections rapidly and accurately.

Animals ; Antibodies, Viral ; blood ; Bocavirus ; genetics ; immunology ; Capsid Proteins ; genetics ; immunology ; Child, Preschool ; Female ; Gene Expression ; Humans ; Infant ; Male ; Parvoviridae Infections ; blood ; diagnosis ; immunology ; virology ; Recombinant Proteins ; genetics ; immunology ; Spodoptera

OBJECTIVETo study the clinical and laboratory features of the mild and severe hand-foot-mouth diseases (HFMD) in Shenzhen in 2008.

METHODS145 cases were observed in East-Lake Hospital and Shenzhen Children's Hospital. Of the 145 cases, 124 mild cases and 21 severe cases were involved.All the clinical data and laboratory findings were collected and summarized. After collection of the acute and convalescent consecutive stools and peripheral bloods from the patients with HFMDI, EV71 genes were amplified from these samples by RT-PCR. Enterovirus 71 were cultured and isolated using Vero cell line and R&D cell line.

RESULTSThe WBC counts and blood glucose levels of the severe cases were significantly elevated, but the ages of the severe ones significantly decreased compared with those of the mild cases (P < 0.05). EV71 genes could be detected by RT-PCR with 35% positive rate in mild cases and 67% in severe cases. The EV71 gene detection rate of the severe cases was significantly increased in contrast to that of the mild ones. The EV71 were isolated and cultured from the stools of 9 patients, one specimens from the dead's stool. Two severe cases died of neurogenic pulmonary edema and brain-stem encephalitis.

CONCLUSIONSEV71 mainly contributes to HFMD and is responsible for death of some severe cases. High fever, less rash, elevated white blood cell counts and blood glucose concentrations as well as age less than 4 years old should be used for prediction of severe cases.

Adult ; Blood Glucose ; physiology ; Child ; Enterovirus ; isolation & purification ; Enterovirus Infections ; blood ; pathology ; Female ; Hand, Foot and Mouth Disease ; blood ; pathology ; virology ; Humans ; Laboratories ; Leukocyte Count ; statistics & numerical data ; Male ; Reverse Transcriptase Polymerase Chain Reaction ; Severity of Illness Index

OBJECTIVE： To establish HPLC fingerprint of Mongolian medicine Ramulus Tamarix， and to conduct similarity evaluation and cluster analysis. METHODS： HPLC method was adopted. The determination was performed on Agilent ODS C18 column with mobile phase consisted of methanol-water （gradient elution） at the flow rate of 1.0 mL/min. The detection wavelength was set at 335 nm， and column temperature was 30 ℃. The sample size was 10 μL. Using quercetin peak as reference， HPLC fingerprint of 10 batches of medicinal sample were drawn. Similarity evaluation was conducted by using Similarity Evaluation System for Chromatographic Fingerprint of TCM （2004 A） so as to confirm common peak. SPSS 17.0 software was used for cluster analysis of those samples. RESULTS： There were 13 common peaks in HPLC fingerprint of 10 batches of samples， and the similarities of them were all greater than 0.95； 2 common chromatographic peaks of quercetin and kaempferol were identified. Results of cluster analysis showed that 10 batches of samples were classified into 2 classes. S8 was clustered into one class； other were clustered into another class. CONCLUSIONS： Established HPLC fingerprint and results of similarity evaluation and cluster analysis can provide reference for quality control of R. Tamarix.

OBJECTIVE To establish the fingerp rint of decoction pi eces and dispensing granules of Gardenia jasminoides ，to determine the contents of 6 components，so as to evaluate its quality combined with chemical pattern recognition. METHODS High performance liquid chromatography （HPLC）was used. Using geniposide as the reference ，Similarity Evaluation System for Chromatographic Fingerprint of TCM （2012 edition）was used to draw the fingerprints of 20 batches of G. jasminoides decoction pieces and 10 batches of G. jasminoides dispensing granules. Similarity evaluation and common peaks identification were conducted. The same HPLC method was adopted to determine the contents of deacetyl asperulosidic acid methyl ester ，geniposide， picrocrocin，rutin，crocin-Ⅰ and crocin- Ⅱ. ORIGIN 9.1 software was used for hierarchical clustering analysis ，and SIMCA 16.0 software was used for principal component analysis （PCA） and partial least squares-discriminant analysis. The differential components affecting the quality of decoction pieces and dispensing granules were screened by taking the variable importance in projection（VIP）value＞1 as the standard. RESULTS There were 24 common peaks for both 20 batches of G. jasminoides decoction piece and 10 batches of G. jasminoides dispensing granules ；a total of 22 common peaks were found in the fingerprints of 30 batches of samples ，and the similarity was not lower than 0.96；six common peaks were identified ，i.e. deacetyl asperulosidic acid methyl ester （peak 2），geniposide（peak 6），picrocrocin（peak 9），rutin（peak 11），crocin-Ⅰ（peak 15），crocin-Ⅱ（peak 17）. Average contents of above 6 components in G. jasminoides decoction pieces were 1.04，57.00，1.30，1.03，9.63 and 0.99 mg/g， respectively；those of G. jasmin oides dispensing granules were 0.96，17.04，0.37，0.27，0.73 and 0.04 mg/g，respectively. PCA results showed that G. jasminoides decoction pieces and G. jasminoides dispensing granules were clustered into respective one category ，which was consistent with results of cluster analysis. There were 9 common peaks with VIP value ＞1， which were 16，14，3，17（crocin-Ⅱ），15（crocin-Ⅰ），18， 22， 2 （deacetyl asperulosidic acid methyl ester） and 21. CONCLUSIONS The estab lished fingerprint and content determination method are simple and reproducible. Combined with chemical pattern recognition ，it can be used to evaluate the quality of decoction pieces and dispensing granules of G. jasminoides . Nine corresponding components represented by peak 16 and so on are the differential components that affect the quality of them.

OBJECTIVE: To explore the genotype mutation characteristics of patients with glucose-6-phosphate dehydrogenase(G6PD) deficiency in Wuhan. METHODS: A total of 1 321 neonates with positive screening and outpatients were received G6PD mutation detection, 12 kinds of common G6PD mutation in Chinese people was detected by using multicolor melting curve analysis (MMCA) method, for those with negative results, the enzyme activity and clinical information were analyzed, sequencing was recommended after informed consent when it is necessary. RESULTS: Among 1321 patients, a total of 768 mutations were detected out, with a detection rate of 58.1%. A total of 18 types of G6PD genotypes were identified, including c.1388G>A, c.1376G>T, c.95G>A, c.1024C>T, c.871G>A, c.392G>T, c.487G>A, c.1360C>T, c.1004C>A, c.517T>C, c.592C>T, c.94C>G, c.152C>T, c.320A>G, c.1028A>G, c.1316G>A, c.1327G>C and c.1376G>C, including 683 male hemizygotes, 3 female homozygotes, 80 female heterozygotes and 2 female compound heterozygous. CONCLUSION A total of 18 types of G6PD mutations are identified in the reaserch, and c.94C>G, c.1028A>G and c.1327G>C are first reported in Chinese population. The most common G6PD mutation types in Wuhan are c.1388G>A, c.1376G>T, c.95G>A.
Asians/genetics* ; Female ; Genotype ; Glucosephosphate Dehydrogenase/genetics* ; Glucosephosphate Dehydrogenase Deficiency/genetics* ; Heterozygote ; Humans ; Infant, Newborn ; Male ; Mutation