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.

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

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.

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 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

OBJECTIVETo study the expression of muscarinic receptor M(3) gene in peripheral blood lymphocytes of workers exposed to organophosphorus pesticides (OPPs) and to explore its role in the adverse effects of OPPs.

METHODSThe lymphocytes of peripheral blood from 33 workers exposed to dimethoate and 15 control people were isolated and treated with saline and dimethoate respectively in vitro. RT-PCR technique was used to determine M(3) gene expression. Basal and inducible gene expression levels were measured.

RESULTSThere was no significant difference in basal gene expression level between exposed group and control group, while the inducible gene expression level was significantly higher in exposure group (1.92 +/- 1.07) than in control group (1.22 +/- 0.19) and basal level (1.49 +/- 0.45, P < 0.05). No differences in basal and inducible gene expression level were found between male and female people in both exposed and control group. The level of inducible M(3) gene expression increased with the increase in length of exposure time [< 5 a: (1.69 +/- 0.95), 5 - 25 a: (1.91 +/- 1.03), > 25 a: (2.09 +/- 1.25), the latter was significantly different from that of < 5 a (P < 0.05)].

CONCLUSIONAfter long-term exposure to OPPs, the basal M(3) receptor gene expression level in the exposed workers did not show any difference from the control group, but the inducible gene expression level (treated with dimethoate in vitro) was increased and related to the extent of exposure to dimethoate.

Dimethoate ; blood ; poisoning ; Female ; Gene Expression ; Humans ; Insecticides ; blood ; poisoning ; Lymphocytes ; cytology ; metabolism ; Male ; Occupational Exposure ; analysis ; RNA, Messenger ; genetics ; metabolism ; Receptor, Muscarinic M3 ; genetics ; Reverse Transcriptase Polymerase Chain Reaction

AIMTo study the effect of volume expansion by 0.9% and 1.8% sodium solution on cardiac-renal reflex activity and the role of cardiac-renal reflex in the regulation of integrated function.

METHODS18 health pentobarbital-anaesthetized rabbits were divided evenly into 2 groups at random, bilateral sino-aortic denervation, intubated via right jugular vein to monitor CVP, left renal nerve separated and ending sectioned to record ERSNA, bilateral ureter intubated to collect urine, right femoral intubated to get blood sample. 15% of whole body blood volume of 0.9% and 1.8% sodium solution were injected via jugular vein 10 ml per minute respectively. The CVP, ERSNA, bilateral urine volume and coefficient of sodium excretion were measured before treated, during treated, one minute, five minutes and ten minutes after treated.

RESULTSVolume expansion by 0.9% and 1.8% sodium solution respectively resulted in the increase of CVP by 64.00% +/- 15.56% and 77.00% +/- 23.74%; the decrease of the frequency of ERSNA by 44.00% +/- 13.64% and 63.00% +/- 12.49%, the average burst time of ERSNA by 37.00% +/- 16.49% and 31.00% +/- 10.69%, the increase of average interval of ERSNA bursts by 60.00% +/- 18.38% and 68.00% +/- 27.04%; the increase of urine volume by 158.00% +/- 28.10% and 640.00% +/- 155.39% in left kidney, 192.00% +/- 32.26% and 1343.00% +/- 429.95% in the right; the increase of coefficient of sodium excretion by 132.00% +/- 35.23% and 376.00% +/- 121.72% in the left, 300.00% +/- 76.99% and 856.00% +/- 261.48% in the right.

CONCLUSIONVolume expansion by different solution stimulate the receptors of cardiopulmonary and regulate the water and sodium excretion of the kidney by the cardiac-renal reflex to modulate the stabilization of blood volume.

Animals ; Blood Volume ; drug effects ; physiology ; Central Venous Pressure ; Heart ; drug effects ; innervation ; Kidney ; drug effects ; innervation ; Rabbits ; Reflex ; Saline Solution, Hypertonic ; pharmacology

OBJECTIVETo survey the incidence of acute febrile reaction in elderly patients receiving intravenous zoledronic acid for osteoporosis and identify the related factors.

METHODSThirty-eight elderly patients with osteoporosis were hospitalized and treated with intravenous infusion of 5 mg zoledronic acid in 2010. The incidence of acute fever reaction was observed in these patients , and the time of fever onset, duration, average maximum temperature, and antipyretic drug used were recorded. The patients with and without acute febrile reaction were compared for age, duration of osteoporosis, sex ratio, use of parathyroid hormone before zoledronic acid treatment, β-fragment of collagen breakdown, calcitonin, osteocalcin, serum calcium, and use of anti-osteoporosis drugs before the treatment.

RESULTSAcute fever reaction occurred in 12 (31.6%) of the patients. Two of the patients had fever on the day of zoledronic acid treatment, and the other patients developed fever after the first day of treatment, with a mean duration of 1 day and a maximum temperature of (38.5∓0.84) degrees celsius;. The fever was treated with a mean of 3.55∓1.21 pseudoephedrine tablets. The patients with fever showed significantly higher parathyroid hormone levels before treatment than those without fever (P<0.05); osteocalcin, calcitonin, β-fragment of collagen breakdown, or serum calcium showed no significant difference between the two groups.

CONCLUSIONAcute febrile reaction, often moderate and transient, is common in elderly patients receiving intravenous zoledronic acid for osteoporosis, and its occurrence is possibly associated with parathyroid hormone levels before the treatment.

Aged ; Aged, 80 and over ; Bone Density Conservation Agents ; administration & dosage ; adverse effects ; China ; epidemiology ; Diphosphonates ; administration & dosage ; adverse effects ; Female ; Fever ; chemically induced ; Humans ; Imidazoles ; administration & dosage ; adverse effects ; Incidence ; Infusions, Intravenous ; Male ; Osteoporosis ; drug therapy ; Parathyroid Hormone ; blood