AIMTo study the function of c-Jun N-terminal kinase 3 (JNK3) in the process of ischemic/reperfused heart injury and the mechanism underlying the protective action of magnesium lithospermate B (MTB), a bioactive compound isolated from Danshen.

METHODSBy in situ hybridization, JNK3 mRNA was detected in the ventricular preparations of the Langendorff ischemic/reperfused rat heart. The inhibitory effect of MTB on the expression of JNK3 mRNA was also investigated.

RESULTSThe purple and blue hybridization signals were located in the cytoplasm of the cardiomyocytes, which were weaker in the non-perfused hearts and stronger in the hearts encountered 30 min of ischemia and 30 min of reperfusion. Image analysis showed that the expression of JNK3 mRNA in the cardiomyocytes increased after 30 min of ischemia and 30 min of reperfusion, which showed significant difference compared with that in the cardiomyocytes of the non-perfused heart and the control heart (P < 0.05). Treatment with of 0.1, 1 and 10 mumol.L-1 MTB abolished the elevation of JNK3 mRNA expression in the ischemic/reperfused heart (P < 0.05).

CONCLUSIONJNK3 may be another component in the signal transduction pathway of ischemia/reperfusion induced cardiomyocyte apoptosis. MTB may protect the heart from ischemia/reperfusion injury by reducing apoptosis through inhibition of the JNK3 activity.

Animals ; Cardiotonic Agents ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; Male ; Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; genetics ; metabolism ; Myocardial Ischemia ; complications ; Myocardial Reperfusion Injury ; enzymology ; etiology ; pathology ; Myocytes, Cardiac ; drug effects ; enzymology ; RNA, Messenger ; biosynthesis ; drug effects ; Rats ; Rats, Sprague-Dawley

OBJECTIVETo elucidate the role of let-7a-mediated gene regulation in the pathogenesis of lung cancer.

METHODSTwo template DNA sequences were designed based on hsa-let-7a sequence in miRBase database. The let-7a expression construct and a control plasmid, namely pGenesil-let-7a and pGenesil-control, respectively, were generated by cloning the annealed oligonucleotides into pGenesil-1 and then transfected into A549 cells, which were selected by G418 to establish the lung cancer cell lines stably expressing let-7a-GFP and control-GFP. The living cells were counted by MTT assay and cell growth curves were drawn to analyze the cell proliferation. The k-Ras mRNA level was assessed by semi-quantitative RT-PCR, and the expression of k-Ras protein was determined by Western blotting and immunocytochemistry.

RESULTSThe recombinant vectors were verified by sequencing. The cell growth curves indicated that the proliferation of the cells transfected with pGenesil- let-7a were inhibited significantly compared with that of cells transfected with pGenesil-control and A549 cells. Semi- quantitative RT-PCR analysis showed that the levels of k-Ras mRNA almost remained unchanged in cells with or without the treatments. Western blotting and immunocytochemistry demonstrated a significant decrease of k-Ras protein levels in cells transfected with pGenesil-let-7a, but not in cells transfected with pGenesil-control, when compared to A549 cells.

CONCLUSIONlet-7a over-expression represses the expression of k-Ras protein and significantly inhibits the growth of lung cancer cells.

Cell Line, Tumor ; Gene Expression Regulation, Neoplastic ; Genetic Vectors ; Humans ; Lung Neoplasms ; metabolism ; pathology ; MicroRNAs ; genetics ; metabolism ; Plasmids ; Proto-Oncogene Proteins p21(ras) ; genetics ; metabolism ; RNA, Messenger ; genetics ; Transfection

OBJECTIVETo study molecular epidemiology of norovirus (NV) infections, stool specimens collected from children with acute diarrhea were tested by TaqMan real-time reverse transcription polymerase chain reaction (RT-PCR) for the viral specific nucleic acid segments.

METHODSFecal samples from a total of 1260 children who had watery diarrhea seen from December 2006 to December 2007 in Guangzhou were analyzed by real-time RT-PCR. The primers and probes used for rapid detection and typing of NV strain target NV sequences were at the ORF1-ORF2 junction, a highly conserved region of the NoV genome. The positive specimens were determined by nested PCR and sequenced.

RESULTSTotally 257 specimens were positive for NV with a positive rate of 20.40%. Shedding of NV type GI was detected in 6.90%, type GII in 16.98% respectively, while the positive number of mixed infection with GI and GII was 44. Of the NV strains that were cloned and sequenced, GI was GI-3, GI-2 and GI-4 detected in positive specimens respectively; meanwhile, GII-4 was most commonly seen in genome II, followed by GII-3 and GII-7. In addition, the average age of children infected with NV was less than 2 years. An epidemic occurred during the winter and early spring (December through the next March).

CONCLUSIONNV was one of the important pathogens for acute diarrhea among children in Guangzhou, which suggested GII-4 was the prevalent strain.

Caliciviridae Infections ; epidemiology ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; etiology ; virology ; Feces ; virology ; Humans ; Infant ; Molecular Epidemiology ; Norovirus ; classification ; genetics ; RNA, Viral ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

OBJECTIVETo detect the high-risk human papillomavirus (HPV) infectious condition in women with abnormal cytology and evaluate its values in the screening of high grade squamous intraepithelial lesion.

METHODSWe used hybrid capture 2 (hc2) method to examine 949 patients with abnormal cervical cytology results [ > or =atypical squamous cells of undetermined significance (ASC-US) according to the 2001 The Bethesda System diagnosis criteria]. All subjects also received colposcopy for tissue studies.

RESULTSAmong 949 patients with abnormal cytology, the diagnoses of atypical squamous cells (ASC), low grade squamous intraepithelial lesion (LSIL), and high grade squamous intraepithelial lesion (HSIL) were made in 432, 310, and 207 patients, respectively. The high-risk HPV positive rate in ASC, LSIL, and HSIL were 40.3%, 44.8%, and 89.4%, respectively. The numbers of patients with pathologically confirmed results of negative intraepithelial lesion or malignancy (NILM), cervical intraepithelial neoplasia 1, 2, 3 (CIN 1, 2, 3), and squamous cell carcinoma (SCC) were 335, 388, 118, 101, and 7, and the high-risk HPV positive rate was 17.3%, 66.2%, 92.4%, 97.0%, and 100%, respectively. Among patients with atypical squamous cells of undetermined significance (ASC-US), rate of HSIL in high-risk HPV positive group and negative group were 10.2% and 0.8%, respectively (P < 0.01). In screening HSIL, the sensitivities of cytology [ > or = ASC cannot exclude HSIL (ASC-H)] and cytology ( > or = ASC-H) plus high-risk HPV testing were 0.925 and 0.991, and the specificities were 0.510 and 0.748, respectively (P < 0.01). Sensitivitives of cytology ( > or = LSIL) and cytology (> or = LSIL) plus high risk HPV in detecting HSIL were 0.898 and 0.982, respectively, while the specificitives were 0. 567 and 0.779, respectively (P < 0.01).

CONCLUSIONSThe positive rate of high-risk HPV increases with the gravity of cervical lesions. In patients with abnormal cervical cytology, high-risk HPV testing can improve the sensitivity and specificity in the screening of HSIL.

Carcinoma, Squamous Cell ; diagnosis ; pathology ; virology ; Cervical Intraepithelial Neoplasia ; diagnosis ; pathology ; virology ; Female ; Humans ; Papillomaviridae ; genetics ; isolation & purification ; Papillomavirus Infections ; diagnosis ; pathology ; virology ; Risk Assessment ; Uterine Cervical Neoplasms ; diagnosis ; pathology ; virology

A series of adefovir mono-L-amino acid esters, mono non-steroidal anti-inflammatory drugs carboxylic ester prodrugs with more potent anti-HBV activity and lower nephrotoxicity were designed and synthesized. Adefovir bis (L-amino acid) ester was used as lead compound, according to pathological and pharmacological findings that non-steroidal anti-inflammatory drugs can effectively inhibit the organic anion transporter 1 (hOAT1)-mediated adefovir phosphonic acid pairs of anion transport across tubular basement membrane thereby reducing the nephrotoxicity of adefovir. Flatten design principle was used to introducing non-steroidal anti-inflammatory drugs structural fragments to design and synthesize target adefovir mixture ester prodrugs. HepG2 2.2.15 cell line was used as in vitro anti-HBV activity evaluation model. Five compounds exhibited antiviral activity, and compound 18 showed the most potent anti-HBV activity and relatively high selective index (EC50 3.92 micromol L(-1), SI 9.97). HK-2 cell line was used as in vitro model to evaluate nephrotoxicity. Results suggested the target compounds have lower cytotoxicity than the positive control. Moreover, by analyzing the primary structure and activity relationship of these compounds, it could suggest that mono-L-amino acid ester, mono non-steroidal anti-inflammatory drugs carboxylic ester prodrugs strategy has significant potential in the acyclic nucleoside phosphonates prodrug design.
Adenine ; analogs & derivatives ; chemical synthesis ; chemistry ; pharmacology ; Amino Acids ; chemical synthesis ; chemistry ; pharmacology ; Anti-Inflammatory Agents, Non-Steroidal ; chemical synthesis ; chemistry ; pharmacology ; Antiviral Agents ; chemical synthesis ; chemistry ; pharmacology ; Carboxylic Acids ; chemistry ; pharmacology ; Cell Survival ; drug effects ; Drug Design ; Hep G2 Cells ; drug effects ; Humans ; Kidney Tubules, Proximal ; cytology ; metabolism ; L-Lactate Dehydrogenase ; metabolism ; Molecular Structure ; Organophosphonates ; chemical synthesis ; chemistry ; pharmacology ; Prodrugs ; chemical synthesis ; chemistry ; pharmacology

Objective To explore the clinical application value of dual-energy CT angiography (DECTA) for head and cervical vessel diseases. Methods The imaging data and diagnosis results of 146 patients suspected as having head and cervical vascular diseases and underwent DECTA in our hospital from February 2009 to August 2009 were analyzed, retrospectively and compared. Sixteen of them with positive results were also performed whole cerebral vessels DSA. Results Seventy-seven patients (52.74%) were found to have head and cervical vessel diseases under DECTA; 17 patients were diagnosed as having intracranial aneurysm, 13 with carotid artherosclerosis and stenosis, 13 with intracranial and vertebral artery stenosis, 9 with intracranial arteriovenous malformation (AVM), 7 with internal carotid artery (ICA) or vertebral artery occlusion, 4 with neck-face AVM, 3 with Moyamoya disease, 3 with intracranial tumors, 2 with cerebral infarction, 2 with intracranial phlebothrombosis, 1 without imaging in bilateral ICA, 1 with Sturg-Webber syndrome and 1 with primitive trigeminal artery.DSA results were concordant with the DECTA results by finding 15 of 16 patients (93.75%) with head and cervical vessel diseases; 8 patients were detected as having intracranial aneurysm, 4 with ICA or vertebral artery occlusion, 1 with carotid artherosclerosis and stegnosis, 1 with intracranial AVM and 1 with neck-face AVM. The one that DSA did not find any tumors was noted as having 2 microaneurysms by CTA. Conclusion The head and cervical vascular diseases and the feeding artery of tumor can be demonstrated distinctly with DECTA with high diagnosis accuracy and sensitivity.

Objective To evaluate the effects and mechanisms of the mieroscrew implant anchorage (MIA) combined with multi-loop edgewise arch wire (MEAW) technique in the treatment of skeletal Class Ⅱ. Adult patients. Methods Eleven adult patients with skeletal Class Ⅱ high-angle maloeelusions were treated with fixed appliances. The spaces were closed by the springs from the MIA to the hook on the arehwire. The height of the hook and the direction of the force were different according to the intrusion and retraction of upper anterior teeth. In the finishing stage, MEAW technique and modified class Ⅱ elastics (from the first loop of MEAW to the MIA) were used for final detailing. Cephalometric analysis was used to evaluate the effect after treatment. Results After treatment, the decrease of SNA, ANB and FMA were ( 2. 86 ± 1.05 )° , ( 2. 82±0. 96 )° and ( 2.95 ± 1.35 )° , respectively. The torque control of upper anterior teeth was good. The protrusion of lower incisors and the molar extrusion were avoided. The upper molars were moved distally by ( 3. 00 ± 2. 19) nun. Conclusions The treatment of adult patients with skeletal Class Ⅱ high angle maloeelusions with MIA and MEAW technique could not only improve the facial esthetics but also avoided the common side effects of traditional Class Ⅱ elastics .

OBJECTIVEGenetic factors are thought to be involved in the development of vitiligo. The aim of this study is to explore the possible genetic model of vitiligo by analyzing the genetic characteristics of 815 patients from Zhejiang province.

METHODSData for 815 patients with vitiligo together with their first- and second-degree relatives were obtained using a standardized questionnaire. All these information was requested to confirm the answers about family history in order to reduce the possibility of 'recall' bias. The 815 probands would include 411 (50.43%) males and 404 (49.57%) females with a varied age from 2 months to 71 years old. Since the information on general prevalence of vitiligo in this area was absent, a control group was set up to facilitate the calculations of heritability degree. 468 persons of the control group were from non-vitiligo population with a sex ratio of 241(male): 227(female) with varied age of 4 months to 80 years old. Both gender and age were comparable between the vitiligo and the control population. The inheritance pattern estimation, heritability calculation and complex segregation analysis were performed with Penrose method, Falconer regression method and SAGE-REGTL program.

RESULTSIn 815 vitiligo probands, 128 had and 687 had not family histories, with a heritability rate of 15.7%. The vitiligo prevalence in proband's first degree relatives was 2.580%, higher than the prevalence of 0.618% in second degree relatives, and both of them were higher than general prevalence: 0.192%. By Penrose method, the rates on different catagories were as follows: sibling prevalence rates s = 0.080 18; population prevalence rate q = 0.001 92; s/q = 41.76. The ratio of s/q did not approach 1/2q (260.42) or 1/4q (130.21), but approached 1/square root of q(22.82), suggesting vitiligo was consistent with a mode of polygenic inheritance. Using Falconer's method, heritabilities of vitiligo in first-and second degree relatives of probands were 59.61% (95% confidence interval 65.37-53.84) and 55.20% (95% confidence interval 43.88-66.52), respectively. The weighted average of heritability in all relatives was 58.7% (95% confidence interval 53.56-63.83). The results of complex segregation analysis suggested that major gene model including the Mendelian dominant, recessive and additive hypotheses were not rejected (P > 0.05). Purely environmental model and no transmission model were rejected at a 0. 001 significance level. According to AIC, Mendelian dominant inheritance was the best-fitted hypothesis.

CONCLUSIONGenetic factors played an important role in the occurrence of vitiligo, and the genetic model of vitiligo could serve as the polygenetic or multifactorial inheritance with major gene trait.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Child, Preschool ; China ; epidemiology ; Female ; Humans ; Infant ; Male ; Middle Aged ; Models, Genetic ; Vitiligo ; epidemiology ; genetics ; Young Adult

Objective To study the pharmacokineties and bioequiva-lenee of two demostic cefprozil tablets.Methods Twenty healthy volun-teers were randomized into two groups,whose plasma concentrations of cis and trans isomers of eefprozil were determined at different time after single oral dose of 1 g cefprozil tablets by own control by HPLC method.The pharmacokinetic pammeters were computed and which were experi-enced variance analysis.Results Gis cefprozil:the main pharmaeoki-netic parameters of refe:rence and trial cefprozil tablets were as follows:t_(max) were(2.4±1.0),(2.3±0.8)h;C_(max) were(15.4±3.5),(14.8±2.8)μg·mL~(-1);t_(1/2) were(1.4±0.1),(1.4±0.1)h;MRT were (3.4±0.6),(3.4±0.6)h;AUC_(0-t) were(61.2±10.6),(60.3±11.4)μg·h·mL~(-1);AUC_(0-∞) were(64.7±11.2),(64.5±11.7)μg·h·mL~(-1),respectively.Trans cefprozil:the relative bioavailability of the trial preparation was(99.3±14.5)%.Trans cefprozil:the main pharmacokinetic parameters of reference and trial cefprozil tablets were as follows:t_(max) were(2.4±0.9),(2.4±0.9)h;C_(max) were(1.6±0.3),(1.5±0.3)μg·mL~(-1);t_(1/2) were(1.2±0.2),(1.5±0.6)h;MRT were(3.2±0.7),(3.2±0.7)h;AUC_(0-t) were(6.1±1.4),(5.6±1.3)μg·h·mL~(-1);AUC_(0-∞) were(6.1±1.4),(5.6±1.3)μg·h·mL~(-1),respectively.The relative bioavailability of the trial preparation was(94.0±17.5)%.Conclusion The results shows that the trial preparation and the reference prepara-tion of cefprozil were bioequivalent.

OBJECTIVETo investigate the association between vitamin D deficiency and risk of gestational diabetes mellitus (GDM) in pregnant Chinese women.

METHODSA nested case-control study was conducted. Clinical and biochemical data were analyzed for 200 subjects with GDM and 200 subjects with normal glucose tolerance (NGT).

RESULTSThe median (interquartile range) serum 25-hydroxyvitamin D (25OHD) levels were 22.39 (17.67, 29.38) and 25.86 (19.09, 34.88) nmol/L in the GDM and NGT groups, respectively. Rates of 25OHD deficiency or insufficiency were significantly higher in the GDM group than in the NGT group. Subjects with 25OHD levels <25 nmol/L had a 1.8-fold higher risk of GDM compared with subjects with higher vitamin D levels. In the GDM group, serum 25OHD was independently associated with HbA1c and insulin resistance after adjusting for confounding factors. In the NGT group, serum 25OHD was independently associated with fasting plasma glucose and systolic blood pressure after adjusting for maternal age and other confounding factors.

CONCLUSION25OHD insufficiency is very common in Chinese women. Low 25OHD status may be associated with insulin resistance and act as a risk factor for GDM.

Adult ; Asian Continental Ancestry Group ; Diabetes, Gestational ; blood ; epidemiology ; Female ; Humans ; Pregnancy ; Vitamin D ; analogs & derivatives ; blood ; Vitamin D Deficiency ; blood ; epidemiology