OBJECTIVEAim of our study was to compare hematological parameters in Tibetan natives with those in Han migrants living on the Tibet plateau in order to determine the potential effects of age, gender, and ethnicity on hematological response to hypoxia.

METHODSBlood hemoglobin (Hb, g/dl), hematocrit (Hct, %), red blood cells (RBC,10(6)/mm3) were measured in 3 588 healthy Tibetan natives and 3 371 Han migrants ranging in age from 5 to 72 years, living at a mean altitudes of 2 664 m, 3 813 m, 4 525m and 5 226 m.

RESULTSHemoglobin (Hb) concentration analysis was made by multiple regression equations relating hemoglobin to altitude and age. For 2 093 Han males, Hb = 9.612+ 0.001440xaltitude+ 0.06148xage. For 1 948 Tibetan males, Hb =12.202+ 0.000462xaltitude+ 0.02893xage. For 1 278 Han females, Hb = 10.858+ 0.000939xaltitude+ 0.02632xage. For 1 640 Tibetan females, Hb = 11.402+ 0.000626xaltitude+ 0.00412xage. Each of the four equations was statistically significant (P < 0.001), and had variance (r2) of 0.86 or more, indicating that altitude and age accounted for at least 85% of the variation in hemoglobin levels. The coefficients for altitude and for age were higher (P < 0.05) in Han males than in Tibetan males and higher (P < 0.05) in Han females than in Tibetan females. The Tibetan postmenopausal females had higher Hb values than premenopausal females only presented at altitude above 4 000 m while this phenomenon was beginning at altitude of 2 664 m among Han females.

CONCLUSIONWe conclude that gender and increasing age in Tibetans are associated with lower hemoglobin values than those in Han at high altitude, and we speculate that genetic factors seems to be important.

Adolescent ; Adult ; Aged ; Altitude ; Asian Continental Ancestry Group ; Child ; Child, Preschool ; Ethnic Groups ; Female ; Hematocrit ; Hemoglobins ; analysis ; Humans ; Hypoxia ; ethnology ; Male ; Middle Aged ; Tibet ; Transients and Migrants ; Young Adult

The effects of beta-cypermethrin (consisting of alpha-cypermethrin and theta-cypermethrin) on the transient outward potassium current (I(A)) and delayed rectifier potassium current (I(K)) in freshly dissociated hippocampal CA3 neurons of rats were studied using whole-cell patch-clamp technique. The results indicated that alpha-cypermethrin increased the value of I(A) and theta-cypermethrin decreased the value of I(A), though both of them shifted steady activation curve of I(A) towards negative potential. theta-cypermethrin contributed to the inactivation of I(A). The results also showed that alpha-cypermethrin and theta-cypermethrin decreased the value of I(K), and shifted the steady state activation curve of I(K) towards negative potential. Both alpha-cypermethrin and theta-cypermethrin had no obvious effects on the inactivation of I(K). theta-cypermethrin prolonged recovery process of I(K). These results imply that both transient outward potassium channels and delayed rectified potassium channels are the targets of beta-cypermethrin, which may explain the mechanism of toxical effects of beta-cypermethrin on mammalian neurons.
Animals ; CA3 Region, Hippocampal ; cytology ; physiology ; Cells, Cultured ; Female ; Insecticides ; toxicity ; Male ; Neurons ; cytology ; drug effects ; physiology ; Patch-Clamp Techniques ; Potassium Channels, Voltage-Gated ; drug effects ; physiology ; Pyrethrins ; toxicity ; Rats ; Rats, Wistar

To establish a new method of quality evaluation of Traditional Chinese medicine by fingerprint and quantitative analysis of multi-components by single marker method (QAMS). The quality evaluation method was established and validated with Houttuyniae Herba. Chlorogcnic acid was selected as markers of ingredients to establish HPLC fingerprint and internal reference standard to determine the contents of other 6 components (new chlorogcnic acid, cryptochlorogenic acid, rutin, hyperin, isoquercitrin, quercitrin) according to the relative correction factor. At the same time, the seven components were determined by external standard method. The accuracy and feasibility of QAMS was evaluated by comparison of the results between external standard method and QAMS. All tested samples contained the 12 common peaks , 7 of which was verified ,and there was no significant differences between the quantitative results of 7 ingredients of multi-components by single marker method and external standard method in 20 batches. The method of fingerprint combined with QAMS has been verified in Houttuyniae Herba and it is to be a new quality evaluation pattern for Traditional Chinese medicine.
Chromatography, High Pressure Liquid ; methods ; standards ; Drugs, Chinese Herbal ; analysis ; chemistry ; Quality Control ; Reproducibility of Results

OBJECTIVETo construct a replication-incompetent adenovirus vector targeting cancer stem cells by modified touchdown PCR and overlap extension PCR and investigate its infection efficiency in CD133(+) SW480 cells in vitro.

METHODSThe two portions of the fiber gene encoding the Ad5 fiber knob domain with the HI loop deleted were amplified using two pairs of designed primers and then linked by overlap extension PCR. The product obtained was identified by sequencing and inserted into prokaryotic expression vector pEGFP-N1. The product, pEGFP-N1 KNOBδHI, contained a unique EcoRV restriction site in the deleted portion of the sequence encoding the HI loop. The gene sequences of the adenovirus fiber were amplified using both common PCR and overlap extension PCR, then identified by sequencing and inserted into pNEB193, resulting in pNEB-F5. CD133(+) SW480 cells were infected with the generated adenovirus vectors Ad5-GFP and Ad5FHI-GFP to investigate the infection efficiency using fluorescent microscope.

RESULTSThe target fragments of expected sizes were amplified by touchdown PCR and overlap extension PCR, but not by common PCR. Ad5FHI-GFP showed a higher infection efficiency than Ad5-GFP in CD133(+) SW480 cells.

CONCLUSIONCompared with common PCR, touchdown PCR and overlap extension PCR can significantly improve the specificity and efficiency of the PCR products for constructing CD133(+) cancer stem cell-selective adenovirus type 5 vector, which provides carriers for tumor-targeted gene therapy.

AC133 Antigen ; Adenoviridae ; genetics ; Antigens, CD ; Cell Line, Tumor ; Genetic Vectors ; Glycoproteins ; Humans ; Neoplastic Stem Cells ; cytology ; virology ; Peptides ; Plasmids ; genetics ; Polymerase Chain Reaction ; methods

It has been known that the glutamate transmission system and N-methyl-D-aspartate receptor (NMDA-R) were possibly related to anxiety processes. Although anxiety symptom can be relieved by NMDA-R antagonists and partial agonists treatment, the functions of NMDA-R and its subunits in anxiety behaviors remain unclear. We used forebrain specific NR2B over-expression mice to examine whether the increase of NR2B subunit level would induce anxiety behaviors. The results indicated that the juvenile (3-5 months old), middle-aged (8-10 months old) and old (19-22 months old ) NR2B transgenic mice showed no significant difference in open field test and elevated plus maze test as compared with the control mice. Capillary electrophoresis of monoamine neurotransmitter in subregions of forebrain revealed no significant difference between transgenic and control mice of 16-18 months age. These results suggest that the increase of NR2B expression and followed NR1 and NR2A expression augmentations in the forebrain have no significant effect on anxiety-related behaviors in mice.
Animals ; Anxiety ; metabolism ; Mice ; Mice, Transgenic ; Prosencephalon ; metabolism ; Receptors, N-Methyl-D-Aspartate ; metabolism

Objective To establish an HPLC with UV method for determination of remifentanil in human blood.Methods Human blood with internal standard was extracted by butyl chloride.The sample was separated on Zobar SE-CN (4.6 mm × 250 mm,5μm) column with a mobile phase of mixture of acetonitrile,methanol and phosphate buffer,the temperature was set at 30℃ with flow rate of 1.5 mL·min~(-1).UV detection at 210 nm. Results The calibration curves were found to be linear at the range 1-20 ng·mL~(-1) of remifentanil in whole blood with a good correlation (γ=0.956).Conclusion This method is rapid,accurate and sensitive,which is very suitable for investigation of clinical blood concentration of remifentanil.

OBJECTIVETo observe anti-HEV IgG response to vaccination of recombinant antigen fragments and evaluate its protection from Hepatitis E Virus infection in rhesus monkeys (Macaca mulatta).

METHODSTwelve monkeys were divided into three groups and immunized respectively with three different recombinant antigens: namely Ag1 (carboxyl terminal 431 amino acids of ORF2), Ag2 (128aa fragment at the carboxyl terminal of ORF2), and Ag3 (full length ORF3 ligated with two ORF2 fragments encoded by 6743-7126nt and 6287-6404nt). The monkeys were challenged intravenously with fecal suspension from experimentally infected rhesus monkeys, and the other three monkeys served as the placebo group for challenge with HEV. The dynamic changes of the levels of ALT and anti-HEV IgG were examined. Pathological changes of liver tissue were observed by light microscope. Excretion of virus was detected by RT-nPCR.

RESULTSHepatic histopathology of two monkeys in the placebo group was consistent with acute viral hepatitis, and ALT was elevated 3-4 weeks after inoculated with virus, up to 10-20 times higher than normal level. The liver tissue of monkeys immunized with antigen kept normal, ALT in several monkeys elevated mildly, and anti-HEV IgG conversation occurred at 1-2 weeks after vaccination, with the titer reaching 1:12,800. The virus RNA could be detected by RT-nPCR from days 7 to 50 in monkeys of control group, and from days 7 to 21 in vaccinated monkeys after challenged with virus.

CONCLUSIONSThe recombinant antigens could induce the production of anti-HEV IgG, which protected rhesus monkeys from acute Hepatitis symptoms related to HEV infection.

Animals ; Antigens, Viral ; immunology ; Hepatitis E ; prevention & control ; Hepatitis E virus ; immunology ; Immunoglobulin G ; immunology ; Macaca mulatta ; RNA, Viral ; blood ; Recombinant Proteins ; immunology ; Vaccination ; Viral Hepatitis Vaccines ; immunology

Objective To determine the awareness, treatment and control of hypertension and its determinants in the elderly hypertensive patients of Dean nationality so as to explore the associated determinants. Methods This was a cross-sectional, population-based survey. A total of 939 persons aged 60 years and older sampled by stratified sampling method from the elderly population of Dean nationality were examined from July to September 2016. Meanwhile, the data of medical history and examination were collected. Results There were 353 elderly patients with hypertension in the 939 subjects. The awareness, treatment and control of hypertension in the elderly hypertensive patients (EHP), which was 47.3％, 36.5％and 22.1％respectively, were higher than the results of previous national study and had the upward trends with the increasing age (<0.05) . In comparison with the reference group, EHP living in middle economic level area had the higher awareness, treatment and control of hypertension (<0.05) . The results of covariate-adjusted multilevel logistic regression model indicated that there was a cross-level interaction effect between middle economic level area and dyslipidemia on the awareness and treatment of hypertension respectively, and a cross-level interaction effect between middle economic level area and low fat diet on the control of hypertension ( <0.05) . The individual level covariates, age (OR=1.3412, 1.3144, 95％ CI 1.0217-1.7609, 1.0030-1.7225) and diabetes ( OR=2.6735, 2.3078, 95％ CI 1.0721-6.6672, 1.0240-5.2007), had the positive association with the awareness and treatment of hypertension;age (OR=1.4740, 95％CI 1.0034-2.1654), non-drinking (OR=3.2056, 95％CI 1.5554-6.6066) and low salt diet (OR=3.6969, 95％CI 1.7996-7.5946) had the positive association with the control of hypertension. Conclusion The awareness, treatment and control of hypertension in the EHP of Dean nationality were higher than those of the national level, and the dominant factors were economic level, age, dyslipidemia, diabetes, non-drinking, low fat diet and low salt diet, etc.

Objective To observe the therapeutic effect of ketogenic diet therapy for children with intractable epilepsy and its safety.Methods Fifteen patients with intractable epilepsy were treated with ketogenic diet that was modified specifically for Chinese people.The compliance,seizure frequency and side effects were followed up.Results Twelve patients maintained on the treatment for 1 month.Among them,the reduction of seizure frequency in 10 patients exceed 50%.Ten patients maintained on the treatment for 3 months.Among them,the reduction of seizure frequency in 8 patients exceed 50%.Five patients maintained on the treatment for more than 6 months.The reduction of seizure frequency all exceed 50%.The reduction of seizure frequency in 4 patients exceed 90%.The seizures of 3 patients were controlled completely.Ten patients among all cases had various adverse effect,such as nausea,vomiting,diarrhea,constipation,hypoglycemia(nonsymptomatic),hyperlipemia and damage of liver function and so on,which could eliminate by anti-symptomatic treatment.Conclusions Ketogenic diet is effective and safe in Chinese children with intractable epilepsy with modified methods specifically for Chinese.The effect is unrelated with seizure types obviously.

OBJECTIVETo select the peptides that specifically bind human cancer stem cell surface marker CD133 from the Ph.D.-7>(TM) phage peptide library.

METHODSWith a biotinylated extracellular fragment of human cancer stem cell surface marker CD133 as the target protein, the CD133 high-affinity peptides were screened from the phage peptide library by liquid phase panning. The clones with high-binding force with human CD133 were then identified by sandwich ELISA and their single-stranded DNA was extracted to test the specificity by competitive ELISA. The amino acid sequences of the selected peptides derived from the phage DNA sequences were synthesized after sequence alignment analysis, and their capacity of binding with colorectal carcinoma cells was assessed by immunofluorescence technique.

RESULTSAfter 4 rounds of liquid phase selection, the phages capable of specific binding with human CD133 were effectively enriched, with an enrichment ratio of 388 times compared to that at the fourth and first rounds. Thirteen out of the 20 clones from the fourth round of panning were identified as positive clones, among which 11 had identical amino acid sequence of TISWPPR, and 2 had the sequence of STTKLAL, and the former sequence showed a stronger binding specificity to CD133.

CONCLUSIONWe have successfully obtained a peptide that specifically binds human CD133 from the Ph.D.-7(TM) phage peptide library, demonstrating the feasibility of screening small molecule high-affinity polypeptides from phage peptide library by liquid-phase panning.

AC133 Antigen ; Antigens, CD ; metabolism ; Biomarkers, Tumor ; metabolism ; DNA, Single-Stranded ; Glycoproteins ; metabolism ; Humans ; Neoplastic Stem Cells ; metabolism ; Peptide Library ; Peptides ; metabolism ; Protein Binding ; Sequence Analysis, DNA