To construct a secretory-expression vector of antimicrobial peptide Bactenecin 7(Bac7),and identify the secretory-expression product in L.lactis MG1363 and its bioactivity.The splicing primers of regulation elements and Bac7 gene,which designed according to codon usage preferences of L.lactis MG1363,were chemically synthesized,and the overlap-extension PCR method was used to splice the full length of Bac7 gene.Then the Bac7 gene was linked to expression vector pMG36e to construct pMG36e/Bac7 vector,and pMG36e/Bac7 was transformed into L.lactis MG1363 by electrophoration.RT-PCR and Western blot assays were applied to investigate the expression of the Bac7 gene in L.lactis,and bioactivity of Bac7 in culture supernatant of L.lactis was tested with plate-diffusion method.The results showed that the Bac7 gene and its regulation elements was amplified and cloned in the vector pMG36e successfully,The secretory-expressed Bac7 in L.lactis MG1363 harboring pMG36e/Bac7 was identified by Western blot,and it had high bacteriostatic activity against E.coli.These results indicate that the recombinant L.lactis MG1363 could express bioactive Bac7,which lays a foundation for further study of oral administration of a Bac7-secreting L.lactis to treat intestinal bacteria infection.

Nanopore technique is a low-cost tool for single-molecule level analysis without the need of label or amplification. The solid nanopores have been widely used in many fields such as chemistry and life sciences due to their advantages such as high stability, good tolerability, controllable size, and easy for modification. The solid nanopores commonly used in recent years are fabricated using two types of materials: membrane and tube. The membrane materials include silicon nitride, two-dimensional materials, aluminium oxide, and polymer membranes. The tube materials mainly include glass capillary and carbon nanotube. This review summarizes and prospects the research progress of different solid nanopores.

OBJECTIVETo understand the incidence of post-stroke depression (PSD) and the therapeutic effect of acupuncture-moxibustion.

METHODSFive hundred and sixty cases were rating with Hamilton depression scale, and lesion parts, gender, age and property of stroke between the PSD and non-PSD were compared by analysis of variance, chi2 test, and then the patients of PSD were randomly divided into two groups and treated respectively with Prozac-20 and acup-moxibustion at Sishencong (EX-HN 1), Anmian, Neiguan (PC 6), Shenmen (HT 7), Zusanli (ST 36), Sanyinjiao (SP 6), Taichong (LR 3), Zhaohai (KI 6), Shenmai (BL 62).

RESULTSThe total incidence rate of PSD was 43.9%, with no relation to the lesion parts, gender, age and property of stroke (P > 0.05), and there were very significant differences in HAMD scores before and after treatment in the two groups (P < 0.0001), with no significant difference in the effective rate between the two groups (P > 0.05).

CONCLUSIONPSD has a high incidence and influences the recovery of nervous function, and it should be treated at early stage. Acup-moxibustion and Prozac-20 have similar therapeutic effect.

Acupuncture Therapy ; Aged ; Depression ; epidemiology ; therapy ; Female ; Humans ; Incidence ; Male ; Middle Aged ; Moxibustion ; Stroke ; complications

AIMTo study the effects of cryopreservation length on the proliferative potential of hematopoietic cells derived from cord blood.

METHODSUsing Dextran-40 and 10% DMSO as cryoprotectants, separated nuclear cells were stored in liquid nitrogen after they were freezed according programme. One month or 4 months later, they were thawed and expanded in serum-free medium for culture and expansion of hematopoietic cell (SFEM) for 5 weeks. Dynamic results were detected every week.

RESULTSAt the 5th week of expanding, TNC were expanded for 1499.0 +/- 115.6-folds and 1513.0 +/- 110.4-folds, respectively. CD34+ cells and CFCs reached to their highest level at the 2nd week and at the 3rd week. CD34+ cells were expanded for 63.8 +/- 6.1-folds and 62.4 +/- 5.7-folds, respectively. CFCs were expanded for 53.8 +/- 6.3-folds and 54.8 +/- 6.7-folds, respectively. Between the two kinds of cells, statistical significant difference in proliferative potential wasn't detected.

CONCLUSIONIn ideal cryopreservative condition, the cryopreservation length would do not affect the proliferative potential of cord blood hematopoietic cells.

Cell Proliferation ; Cell Survival ; Cells, Cultured ; Cryopreservation ; methods ; Fetal Blood ; cytology ; Hematopoietic Stem Cells ; cytology ; Humans ; Time Factors

Female ; Humans ; Infant ; Propionic Acidemia ; diagnosis

OBJECTIVETo explore the change in the distensibility of large arteries and its influencing factors in elderly patients with essential hypertension.

METHODSAutomatic measuring system for pulse wave velocity (PWV) was applied to examine carotid-femoral PWV as an index reflecting distensibility of large arteries. 118 hypertensive patients aged 64 - 83 (mean age 67.12 +/- 10.26) years were included in the study. Of them, 87 were males and 31 were females.

RESULTSPWV of 118 hypertensive patients increased with increasing age (P < 0.001). Multivariate regressive analysis demonstrated that age and systolic blood pressure had the close relationship with PWV (P < 0.001).

CONCLUSIONHypertension of the elderly could cause reduction of distensibility of large arteries. Age and systolic blood pressure had the close relationship with distensibility of large arteries in elderly patients with essential hypertension.

Age Factors ; Aged ; Aged, 80 and over ; Blood Flow Velocity ; Carotid Arteries ; physiopathology ; Elasticity ; Female ; Femoral Artery ; physiopathology ; Hemodynamics ; Humans ; Hypertension ; physiopathology ; Male ; Middle Aged ; Pulse

OBJECTIVETo promote a new kind of alternative needle of filiform-fire needle with fire needle.

METHODSThe common three-headed fire needle, thick fire needle, less-thick fire needle, fine fire needle, single-filiform needle and three-filiform-needle were used by 20 medical staffs, and the time of heating needles was recorded. The advantages of using filiform needles compared with common fire needles were evaluated.

RESULTSThe time of heating single and three-filiform needles was shorter than the other four common fire needles (all P < 0.01).

CONCLUSIONFiliform needle is better than the common fire needle in terms of production process,materials, and heating time, etc. Therefore, filiform needles can replace common fire needles for pricking.

Acupuncture Therapy ; instrumentation ; Adult ; Equipment Design ; Female ; Humans ; Male ; Middle Aged ; Needles ; Young Adult

Adolescent ; Adult ; Aged ; DNA Mutational Analysis ; DNA, Viral ; genetics ; Drug Resistance, Viral ; drug effects ; genetics ; Female ; Hepatitis B virus ; drug effects ; genetics ; isolation & purification ; Hepatitis B, Chronic ; genetics ; virology ; Humans ; Male ; Middle Aged ; Young Adult

OBJECTIVETo investigate the prevalence of vitamin D receptor (VDR) gene polymorphism in pre-menopausal women in Guangzhou and study its relationship with bone mineral density(BMD).

METHODSThe genotypes of VDR gene in 193 per-menopausal women in Guangzhou were analyzed by polymerase chain reaction-restriction fragment length polymorphism. BMD of the lumbar spine, femoral neck, greater trochanter and Ward's triangle were measured by dual-energy X-ray absorptiometry.

RESULTSIn the 193 subjects, 120 (66.2%) were identified as VDR bb genotype, 64 (33.2%) as Bb, and 9 (4.6%) as BB. The b allele frequencies reached 78.76%, and B allele frequencies was 21.24%. The distribution followed the Hardy-Weinberg equilibrium. No significant difference was found in BMD among the subjects with different genotypes.

CONCLUSIONVDR genotype is not related to BMD, and VDR polymorphism can not be used as a genetic marker for predicting the risk of osteoporosis in pre-menopausal women in Guangzhou.

Absorptiometry, Photon ; Adult ; Analysis of Variance ; Bone Density ; China ; Female ; Gene Frequency ; Genotype ; Humans ; Middle Aged ; Osteoporosis ; genetics ; metabolism ; Polymorphism, Genetic ; Premenopause ; Receptors, Calcitriol ; genetics

OBJECTIVETo study the tolerance of rats induced by 28 day pretreatment with low dosage of dimthoate and the toxic effects challenged by higher dosage of dimethoate, and to investigate the change of M receptor and the mechanism of tolerance formation.

METHODSSD rats were given 25 mg/kg dimethoate daily(sc) while control group was given saline daily instead for 28 days. The activity of whole blood acetylcholinesterase (AChE) was examined. On the 29th day three groups of administrated rats were challenged by saline solution, 50 mg/kg and 100 mg/kg dimethoate, respectively. The density and mRNA level of brain M(1), M(2) receptor were determined. Lymphocytes of peripheral blood were isolated, and basal, inducible M(3) gene expression were measured by RT-PCR.

RESULTSDuring pretreatment, blood AChE activity decreased continually, it reached the lowest on the 13th day. And it decreased more after exposed to higher dosage of dimethoate. Brain AChE activity in the pretreated groups was lower than that in control group and decreased with the increase in challenging dosage. The density of M(1) receptor in negative control, pretreated, and 50, 100 mg/kg challenging groups were 979.15, 856.54, 539.46, 539.14 fmol/mg pro respectively. The change in relative levels of mRNA of M(1) receptor (2.59, 2.47, 2.20, 1.81) were consistent with the density of receptor but the level declined continually as the challenging dosage increased. The density of M(2) receptor were 507.38, 611.11, 548.42, 337.47 fmol/mg pro respectively, which were not obviously affected by pretreatment but decreased as the challenging dosage increased. The change in levels of M(2) receptor mRNA was not obvious. The basal gene expression of M(3) receptor mRNA was not different among all experimental groups while the inducible gene expression decreased with the increase in challenging dosage.

CONCLUSIONLow level dosage of dimethoate could induce animals to tolerate dimethoate toxicity. Reduction of M(1) receptor density which may be induced by the decrease in its gene expression may be the mechanism of tolerance. The change of M(3) receptor mRNA inducible expression in lymphocyte accorded with M(1) receptor mRNA expression in the brain.

Animals ; Brain ; metabolism ; Dimethoate ; toxicity ; Gene Expression ; drug effects ; Insecticides ; toxicity ; Lymphocytes ; metabolism ; Male ; Maximum Tolerated Dose ; RNA, Messenger ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Receptors, Muscarinic ; genetics ; Reverse Transcriptase Polymerase Chain Reaction ; Time Factors