ObjectiveTo observe the effect of insulin detemir combined with acarbose in treatment of patients with type 2 diabetes (T2DM).Methods32 T2DM patients whose blood glucose control was poor receiving twice daily isophane protamine biosynthetic human insulin( Novolin 30R) combined or not combined with oral hypoglycemic drugs,were switched to receive insulin detemir injection once daily and oral acarbose three times daily for 12 weeks.By self-contrasted method,observed blood glucose before and after three meals,before sleeping,glycated hemoglobin ( HbA1c),change of body mass index( BMI),incidence of hypoglycemia,compliance and satisfaction of patients (by questionnaire) before and after treatment.ResultsAfter treatment,the blood glucose before and after three meals,before sleeping and HbA1 c declined sigmifieatly ( t =11.212,14.997,10.863,17.950,11.108,14.034,12.422,22.764,all P ＜0.01 )compared to state before treatment.The rate of hypoglycemia in the day was 3.1％,without nocturnal symptoms of hypoglycemia.BMI had certain declining,but there was no statistical significance (P ＞ 0.05).The compliance and satisfaction of patients were in a higher rate (x2 =10.255,9.143,all P ＜ 0.01 ).ConclusionInsulin detemir combined with acarbose could effectively control the blood glucose,incidence of hypoglycemia,without increasing body mass,achieve good compliance and satisfaction of patients.

The article introduces the current situation of higher medical education of Taiwan Province and makes some brief comments and analyses by analyzing its advantages and disadvantages as well as the effectuation condition of higher education reforming measures.

Objective To investigate the characteristics of high-risk cholecystolithiasis and summarize the experience of laparoscopic chol-ecystectomy for this disease.Methods A retrospective analysis was performed on the clinical data of 1 15 patients with high-risk cholecys-tolithiasis who underwent laparoscopic cholecystectomy in our hospital from October 2008 to March 2012.Results Of the 1 15 patients,47 had stones filling the gallbladder as well as atrophy of the gallbladder and porcelain gallbladder,42 had acute suppurative and gangrenous cholecystitis,20 had stone incarceration in the gallbladder neck,3 had cystic duct stones,2 had cholecystoduodenal fistula,and 1 had Mi-rizzi syndrome;all patients were cured and discharged.Conclusion For patients with high-risk cholecystolithiasis,laparoscopic cholecys-tectomy is feasible,given active preoperative preparation,strengthened perioperative management,and accurate,standard,and skilled sur-gical operation.

Objective To evaluate the effect of tacalcitol on the proliferation,adhesion,migration and c-kit mRNA expression of cultured human epidermal melanocytes.Methods Cultured epidermal melanocytes from the prepuce of adolescent males were treated with various concentrations of tacalcitol.Then,cell proliferation was evaluated by tetrazolium salt (XTT) assay after 24,48 and 72 hours of treatment,adhesive activity by using fibronectin-coated culture plates after 72 hours,migratory activity by Transwell assay using a microporous membrane after 24 hours,and the c-kit mRNA expression was semiquantitatively analyzed by reverse transcription PCR after 72 hours of treatment.Statistical analysis was done by repeated-measure analysis of variance and completely random design analysis of variance.Results As repeated-measure analysis of variance showed,tacalcitol of 10-10,10-9,10-8,10-7 and 10-6 mol/L significantly promoted the proliferation of melanocytes (F =9.47,P ＜ 0.01),with significant differences in the promoting effect among various durations of treatment with different concentrations of tacalcitol (F =14.44,P ＜ 0.01),and with significant interaction effect between drug concentration and treatment duration (F =2.47,P ＜ 0.01).The highest proliferation level was observed in melanocytes treated with tacalcitol of 10-s mol/Lfor 72 hours.There was a significant increase in the adhesion rate of human epidermal melanocytes to fibronectin after treatment with tacalcitol of 10-8-10-7 mol/L for 72 hours (both P ＜ 0.01),number of melanocytes migrating through micropore membranes per high-power field (× 200) after treatment with tacalcitol of 10-9-10-8 mol/L for 24 hours (both P ＜ 0.01),and in the c-kit mRNA expression in melanocytes treated with tacalcitol of 10-9-10-7mol/L for 72 hours (all P ＜ 0.01).Conclusion Tacalcitol can promote melanocytes to proliferate,migrate,express c-kit mRNA,and adhere to fibronectin.

Objective To study the regulatory effect of ethanol extract of glossy privet fruit and its monomer tyrosol on the adhesion and migration of human epidermal melanocytes.Methods Epidermal melanocytes were isolated from human foreskin,and subjected to a primary culture.Mter 3-5 passages,the melanocytes were treated with various concentrations of ethanol extract of glossy privet fruit (0.0375-0.6 mg/ml)and tyrosol (0.125-2 mmol/L) for 24-72 hours.The XTT colorimetric assay was carried out to evaluate the proliferation of melanocytes,fibronectin (FN)-coated culture plates were used to evaluate the adhesion activity of melanocytes,and Transwell assay was conducted to assess the migration activity of melanocytes.Confocal laser microscopy was utilized to observe the structure and distribution of actin cytoskeleton in melanocytes,and cellular fluorescence intensity was determined by a semi-quantitative analysis.Statistical analysis was carried out by using unpaired t test.Results The adhesion activity of melanocytes to FN was significantly enhanced by the ethanol extract of 0.0375-0.6 mg/ml from glossy privet fruit (P ＜ 0.05 or 0.01),and by tyrosol of 0.5-2 mmol/L (P ＜ 0.05 or 0.01).As XTT assay showed,neither the ethanol extract of 0.15 mg/ml nor tyrosol of 2 mmol/L had cytotoxicity or promotive effect on cell proliferation.Hence,0.15 mg/ml and 2 mmol/L were determined as the working concentrations of ethanol extract and tyrosol respectively.The number of cells migrating through micropore membranes per high-power field (× 200) was 43.7 and 51.0 in melanocytes treated with the ethanol extract of 0.15 mg/ml and tyrosol of 2 mmol/L,respectively,significantly higher than that in untreated melanocytes (20.3,both P ＜ 0.01).Compared with untreated melanocytes,those treated with the ethanol extract of 0.15 mg/ml and those with tyrosol of 2 mmol/L showed higher intracellular fluorescence intensity (P ＜ 0.01) and more stress fiber bundles which congregated inside the cell membrane and around the nuclei.Conclusions The ethanol extract of glossy privet fruit and its monomer tyrosol can promote the adhesion and migration of human melanocytes in vitro,likely by promoting the congregation of actin cytoskeleton in melanocytes.

Objective To explore the correlation between serum hs-CRP and β-cell finction in patients with gestational diabetes mellitus(GDM).Methods The levels of hs-CRP in 60 patients with GDM(GDM group)and 30pregnant women with normal glucose tolerance(NGT group)were detected.Insulin resistance was assessed by the homeostasismodel insulin resistance index(HOMA-IR),Insulin secretion by the homeostasis β-cell funetiOn index(HOMA-β).Results The levels of hs-CRP and HOMA-IR were higher in GDM group than in NGT group.There was significant difference between two groups(P＜0.01);HOMA-β was lower in GDM group than in NGT group,there was significant difference between two groups(P＜0.05).The level of hs-CRP was correlated with age,pre-pregnant bodymass-index(BMI),screening BMI,fasting blood glucose(FBG),fasting insulin(Fins),and HOMA-IR(r=0.222,0.649、0.862、0.923、0.935、0.941,P＜0.05 or P＜0.01),but was inversely related with HOMA-β(r=-0.872,P＜0.01).Multiple stepwise regression analysis indicated that HOMA-IR and HOMA-β was the most important effect factors of hs-CRP.Conclusion The level of hs-CRP increased in women with GDM.which was related to insulin resistance(IR)and insulin secretion,and it maybe participate in the pathogenesis of GDM.

Objective To investigate the cerebral vascular autoregulation in patients with mitochondrial encephalomyopathy with lactic acidosis and strokc-like episodes (MELAS) during the remission of stroke-like episodes,including cerebrovascular CO2 reactivity and vascular endothelial function.Methods Twenty-nine MELAS patients confirmed by genetic analysis were recruited in this study. They underwent the examination at least 2 weeks after the onset of last stroke-like episode.Twenty-eight healthy people were collcctcd as healthy controls. Carotid ultrasound and brain magnetic resonance angiogram (MRA) were done to evaluate the cervical and intracranial appearance of large arteries. Evaluation of vascular autoregulation included: (1) the cerebrovascular CO2 reactivity with breath-holding test by transcranial Doppler and calculating breath holding index (BHI),and ( 2 ) flow-mediatcd dilation ( FMD )and nitroglycerin-mediated dilation with ultrasound assessment of humeral artery.Independent-samples t test was done between the results of two groups.Results Carotid ultrasound and cranial MRA revealed no abnormalities in both MELAS patients and healthy controls.The BHI of MELAS patients was significantly decreased than that of normal controls ( 1.36 ± 0.52 vs 1.81 ±0.26,t =- 3.693,P ＜ 0.01 ),and the FMD of MELAS patients was also significantly lower than that of normal controls (11.0％ ±4.8％ vs 15.8％ ±5.8％,t =-3.390,P ＜0.01).Conclusion The function of vascular autoregulation,including cerebrovascular CO2 reactivity and FMD,is impaired in MELAS patients.

Objective To investigate the relationship between the single nucleotide polymorphisms (SNPs) of TLR9 gene and the occurrence of condyloma acuminatum (CA). Methods Peripheral venous blood was obtained from 63 patients with CA and 23 normal human controls with informed consent. DNA was extracted from the blood samples and subjected to the amplification of TLR9 gene by PCR followed by sequence analysis. Results There were 4 SNPs, i.e., SNP1, SNP2, SNP3 and SNP4 at positions 1174, 1635, 1269 and 1724 of the TLR9 gene, respectively. Of these SNPs, SNP1 was located in intron 1, SNP2, SNP3 and SNP4 in exon 2. The registration number is rs352139 for SNP1, rs352140 for SNP2 in NCBI database. SNP3 and SNP4 were newly discovered positions. The frequency at SNP1 position was 0.690 and 0.609 for allele A in the patients and controls, respectively, 0.309 and 0.391 for allele G, respectively (both P > 0.05). No significant difference was observed between the patients and controls in the frequency of allele A or allele G at position SNP2 (0.302 vs. 0.698, 0.369 vs. 0.630, both P > 0.05). There were 4 haplotypes at the SNP1 and SNP2 positions, including AA, AG, GA and GG, with no significant difference in the frequency between the patients and controls (all P> 0.05). Conclusions There are 4 SNPs including SNP1, SNP2, SNP3 and SNP4 in the TLR9 gene in Guangdong Han population. SNP1 and SNP2 appear unrelated to the liability to CA.

Objective To elucidate the genome organization of small deletion mutants of hepatitis B virus(HBV).Methods Amplified the HBV genomes by polymerase chain reaction from the serum of the patients with chronic hepatitis B and cloned the small HBV DNA less than 1 kb,then sequenced and analyzed the gene organization of these small deletion mutants of HBV.Results Totally one hundred and twenty-four small deletion mutants of HBV genomes categorized to sixty-four types were obtained and classified into three groups according to the criteria of the characteristics of gene organization,for example,spliced variants,regular deletion mutants and the deletion mutants with an internal poly (dA).All of these isolated mutants shared some common features as the deletion in coding regions and regulatory elements,66% of the mutants retained the cis elements crucial for the viral replication and encapsidation,while 48% retained the X region.Conclusions Small deletion mutants of HBV are commonly detected in the serum from chronic hepatitis B patients,the characteristic structure of such mutants implies that they might be closely co-related with the pathogenicity of HBV.The exact mechanisms need further study yet.

Objective To investigate the anti-IFN-α effects of the novel protein TSR'r' encodedby the 458 nt-1308 nt spliced variant of hepatitis B virus genome,and to determine its functional domaias.Methods the TSR'r' gene(originated from open reading frame of HBV DNA polymerase,T represents terminal protein region,S represents the Spacer region,R'represents the truncated reverse transcriptase region,and r'represents the truncated RNaseH region)of the 458 nt-1308 nt spliced variant of HBV genome and its deletants were amplified by PCR and were cloned into the pcDNA3.1/HisC vector.The recombinant vector was transfected into Huh7 hepatocytes individually by FuGENE6 transfection reagent,and the expression of the fusion protein was detected by Western blot.Huh7 hepatocytes were co-transfected with p6 16CAT and the recombinant vector harboring either TSR'r'or the related deletant,and treated with IFN-α 2a 48 h post transfection.After 24 h stimulating.the cells were lysed and the intracellular CAT value was calculated.All data were processed with One-way analysis of variance(ANOVA).Resuits Recombinant vectors harboring either the TSR'r'gene or related deletant were constructed successfully,and the fusion proteins were expressed well in Huh7 cells.When Huh7 hepatocytes were co-transfected with p6-16CAT and TSR'r' recombinant.the intracellular CAT values reduced gradually as paralleled with the increasing amount of TSR'r'recombinant.Furthermore,as compared with the empty vector,intracellular CAT values also decreased significantly when the Huh7 cells co-transfected with recombinant harboring TP plus Spacer regions,while any of the other deletants(harboring either TP or Spacer region or neither)showed no significant difference.Conclusion The novel protein encoded by the 458 nt-1308 nt spliced variant of hepatitis B virns genome suppressed the response of Huh7 hepatocytes to IFN-α.and the N-terminal TP plus Spacer region was the functional domain of the protein for anti-IFN-α effects.