A high productive poly ?-glutamic acid(?-PGA) strain PGA-N in a culture medium containing no L-glutamine was isolated from fermentation products.With the following identifications of colony mor-phology,physiological and biochemistry experiments,and genetics,the strain PGA-N was classified as a Bacillus licheniformis.According to the product environment,the base culture medium having no L-glutamine was simulated.In order to enhance the production of the strain PGA-N,the fermentation condi-tions,such as carbon source,nitrogen source,were optimized and the ?-glutamic acid production reached 5.16 g/L after getting the optimum formulation of this culture medium.PGA-N was mutagenized with com-bination of NTG and UV.A mutant PGA-N-C10 was screened which PGA production was increased from 5.16 g/L to 8.82 g/L.The study also investigated the effects of agitation speed on the cell biomass,?-PGA production and the ?-PGA molecular weight.The ?-PGA yield of PGA-N-C10 was as high as 11.00 g/L when the agitation speed was 400 r/min.

Acetic acid bacteria are Gram-negative,obligate aerobic bacteria that have the ability to incompletely oxidize alcohols or sugars to organic acids as end products. The taxonomy of acetic acid bacteria has undergone many changes in the last 30 years. The early classification systems for these bacteria were based on morphological and biochemical characteristics. Today,the acetic acid bacteria are classified as the consensus result of a polyphasic analysis,combining phenotypic,chemotaxonomic and genotypic data. This paper reviewed the polyphasic taxonomy of acetic acid bacteria,mainly introduced the current classification of acetic acid bacteria,then discussed the application of phenotypic,chemotaxonomic and genotypic method in the taxonomy of acetic acid bacteria.

The aim of the present study was to investigate the effects of quercetin on colon contractility and voltage-dependent Ca(2+) channels in the single smooth muscle cell isolated from the proximal colon of guinea-pig and to clarify whether its effect on L-type Ca(2+) current (I(Ca,L)) would be related to its myorelaxing properties. Colon smooth muscle strips were used to take contractile tension recordings. Smooth muscle cells were freshly isolated from the proximal colon of guinea-pig by means of papain treatment. I(Ba,L) (barium instead of calcium as current carrier) was measured by using whole-cell patch-clamp techniques. The results showed that quercetin relaxed colon muscle strips in a concentration-dependent manner and antagonized the contractile effect of acetylcholine and neostigmine. Preincubation with indomethcin [cyclooxygenase (COX) inhibitor] and methylene blue [guanylate cyclase (GC) inhibitor] significantly attenuated the relaxing effect of quercetin, respectively. Quercetin increased I(Ba,L) in a concentration- [EC(50)= (7.59+/-0.38) mumol/L] and voltage-dependent pattern, and shifted the maximum of the current-voltage curve by 10 mV in the depolarizing direction without modifying the threshold potential for Ca(2+) influx. Quercetin shifted the steady-state inactivation curve toward more positive potentials by approximately 3.75 mV without affecting the slope of activation and inactivation curve. H-89 (PKA inhibitor) abolished quercetin-induced I(Ba,L) increase, while cAMP enhanced the quercetin-induced I(Ba,L) increase. The patch-clamp results proved that quercetin increased I(Ba,L) via PKA pathway. It is therefore suggested that the relaxing effect of quercetin attributes to the interaction of GC and COX stimulation, as well as the antagonism effect on acetylcholine, which hierarchically prevails over the increase in the Ca(2+) influx to be expected from I(Ca,L) stimulation.
Animals ; Calcium Channels, L-Type ; metabolism ; Cells, Cultured ; Colon ; drug effects ; Guinea Pigs ; Muscle Contraction ; Myocytes, Smooth Muscle ; drug effects ; Patch-Clamp Techniques ; Quercetin ; pharmacology

OBJECTIVETo develop a RP-HPLC method for the determination of the concentration of hydroxysafflor yellow A in rat plasma, to study the pharmacokinetics of Carthamus tinctorius extration and Naodesheng tablet, and to investigate the effect of other components on the pharmacokinetics of hydroxysafflor yellow A.

METHODThe rats were orally treated with Carthamus tinctorius extration and Naodesheng capsule respectively. Blood samples were collected in heparinized eppendorf tube via the oculi chorioideae vein. Plasma was separated by centrifugation at 10 000 r x min(-1) for 10 min, and two-times methanol in volume was added to deposit proteins. After centrifugation, the upper liquid was transferred to filter. The concentration of hydroxysafflor yellow A in serum was determined by RP-HPLC. The stationary phase was C18, and methanol-acetonitrile-0.7% orthophosphoric acid (26: 2:72) was taken as the mobile phase, A UV detector was used at 403 nm. The pharmacokinetic parameters were calculated with 3p97 program.

RESULTA good linear relationship of hydroxysafflor yellow A was obtained in the range of 0.03 and 2.56 mg x L(-1), the lowest limit of determination was 10 microg x L(-1), and the lowest limit of quantitation was 30 microg x L(-1). The mean recoveries were (99.3 +/- 1.4)%, (92.8 +/- 1.8)%, (98.4 +/- 2.0)% for high, middle, low concentrations of the samples respectively. The plasma concentration-time curves of hydroxysafflor yellow A were fitted with two-compartments model. The AUC)0-t), AUC(0-infinity), C(max) and T(max) of hydroxysafflor yellow A were increased in the Naodesheng group, compared with 50 mg x kg(-1) C. tinctorius extract group.

CONCLUSIONThe HPLC method was selective, accurate and sensitive. The results indicated that the other herbs improved the absorption of hydroxysafflor yellow A and increased the bioavailability of hydroxysafflor yellow A significantly.

Animals ; Biological Availability ; Carthamus tinctorius ; chemistry ; Chalcone ; analogs & derivatives ; pharmacokinetics ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; pharmacokinetics ; Male ; Plant Extracts ; pharmacokinetics ; Quinones ; pharmacokinetics ; Rats ; Rats, Sprague-Dawley

A poly-?-glutamic acid producing strain--BLN-2, was isolated from the soybean products. According to the biochemical characteristics and 16S rRNA, the strain was identified as Bacillus subtilis. Using soybeans as culture, the solid-state fermentation conditions of BLN-2 have been studied. The results showed that the optimal carbon and nitrogen sources of BLN-2 were glucose, fructose, NaNO3 and KNO3, respectively. The orthrogonal experiments showed, when the final concentration of the fructose which was added to the soybean culture was 0.5%, the glucose, NaNO3 and KNO3 final concentraion were 2.0%, the production of ?-PGA was the highest--89.05 g/kg. It is 48.42% higher than other comparable soybean medium under the same conditions.

Objective To observe the Th17 and regulatory T cells(Tr) equilibrium state as well as their changes of tuberculosis patients in six-month's anti-Tuberculosis treatment.Methods Select thirty-two tuberculosis patients received anti-Tuberculosis treatment while thirty-two healthy volunteers as controls.Flow cytometry was used to analyze Th17 and Tr cells in venous blood at the time of pre-therapy,3th,6th month.Results The ratio of Th17 cells in CD4 cells in tuberculosis patients and volunteers were (1.10±0.39)％,(2.50±1.03) ％,(3.90±1.34) ％,(4.50±1.52)％,respectively; the ratio of Tr cells were (9.17±3.26)％,(6.85±2.73)％,(5.46±1.69)％,(4.35±0.86)％,respectively.Conclusion Tuberculosis could make Th17 cells and Tr cells lost their balance,but the immune equilibrium state may gradually recover after anti-tuberculosis.The change of the amount of immune cells was likely to be the reference indexes to observe the progress of tuberculosis and the treatment effect of anti-tuberculosis.

Objective To observe the effects of aerobic training and aerobic combined with resistance training on motor function, like muscle strength,cardiopulmonary endurance and so on,in patients with chronic kidney disease(CKD). Methods From July,2015 to August,2016,60 patients with CKD were randomly divided into control group(A,n=20), aerobic training group(B,n=20)and aerobic combined with resistance training group(C,n=20).Group B per-formed cycle ergometer at 50% peak oxygen uptake(VO2peak)for 30 minutes a time,and group C performed one section of Thera-Band resistance training based on group B, three times a week for twelve weeks.All patients were evaluated with one repetition maximum-upper limb (1 RM-U), one repetition maximum-lower limb (1 RM-L),Cardiopulmonary Exercise Test(CPET),Arm Curl Test(ACT),30-second Chair Stand(CS-30),Six-Min-ute Walk Test(6MWT),and estimated gomerular filtration rate(eGFR)and serum creatinine(sCr)were calculat-ed and recorded before and after training. Results There was no significant difference in all indexes among three groups before training(F<1.841,P>0.05).After training,all indexes improved in groups B and C(t>2.162,P<0.05),and were better in groups B and C than in group A(t>2.132, P<0.05).After training, 1 RM-U, 1 RM-L, VO2peak,ACT, CS-30 and 6MWT were better in group C than in group B(t>2.081,P<0.05). Conclusion Aerobic training could improve the motor function of patients with CKD,and it is more effective combined with resistance training.

The objective of this study was to investigate the effect of different rehydration conditions on recovery of the lyophilized red blood cells (RBC) so as to optimize the RBC rehydration. The different conditions, including different rehydration solution, the rehydration temperature, volume change rate of the lyophilized RBC rehydrated by the vapor firstly, were studied, the recovery rate and change of physiological and biochemical properties of the rehydrated RBC were detected. The results indicated that the solution of 10% (w/v) PVP40 in PBS showed the best effect, and the RBC recovery rate increased with increasing of rehydration temperature, and the optimal temperature of rehydration was at 37 degrees C. Pre-rehydration in condition of vapor could raise the RBC recovery rate, and promote the MCV and RDW to close to index of the fresh RBC, the deformability of the rehydrated RBC was no serious as compared with RBC preserved in conventional condition, but the activity level of ATP, G-6-PD, SOD, 2, 3-DPG of the rehydrated RBC less decreased. It is concluded that the optimal rehydration conditions for lyophilized RBC are pre-rehydration in the 37 degrees C with vapor firstly, PBS + 10% (w/v) PVP40 rehydration solution and rehydration temperature at 37 degrees C, but the protection of RBC membrane needs to be furtherly studied.
Blood Preservation ; methods ; Erythrocyte Count ; Erythrocytes ; Freeze Drying ; methods ; Humans ; Rehydration Solutions ; Temperature

OBJECTIVETo investigate the effect of perioperative application of intestinal probiotics to substitute oral intestinal antimicrobial agents on intestinal flora and immune function in surgical patients with colorectal cancer.

METHODSSixty patients with colorectal cancer undergoing elective laparoscopic radical surgery were randomized to receive preoperative bowel preparation using oral intestinal antimicrobial agents (n=20) or using oral intestinal probiotics (Jinshuangqi Tablets, 2.0 g, 3 times daily) since the fifth day before the operation and at 24 h after the operation for 7 consecutive days. Upon admission and 7 days after the operation, fecal samples and fasting peripheral venous blood were collected from the patients to examine the intestinal flora and serum levels of interleukin-2 (IL-2), IgA, IgG, and IgM, NK cell activity, T lymphocytes subsets CD3(+), CD4(+), CD8(+) and CD4(+)/CD8(+) ratio.

RESULTSAt 7 days after the operation, the patients receiving probiotics showed significantly increased counts of intestinal Bifidobacterium, Lactobacillus, and Enterococcus (P<0.05) and significantly lowered counts of Escherichia coli and Staphylococcus aureus (P<0.05). The serum levels of IL-2, IgA, IgG and IgM as well as CD4(+) cell percentage all increased significantly in probiotics group compared with those in patients with conventional intestinal preparation (P<0.05).

CONCLUSIONSPerioperative application of intestinal probiotics to replace preoperative oral intestinal antimicrobial agents can effectively correct intestinal flora imbalance and improve the immune function of surgical patients with colorectal cancer.

Aged ; Bifidobacterium ; Colorectal Neoplasms ; immunology ; microbiology ; Female ; Humans ; Intestines ; microbiology ; Intraoperative Period ; Male ; Middle Aged ; Premedication ; Probiotics ; therapeutic use ; Prospective Studies ; Single-Blind Method

Objective To examine HIV prevalence and related risk factors among men who have sex with men(MSM) in Chongqing, and to explore the feasibility of using respondent-driven sampling (RDS) in the survey. Methods Based on results from formative research, a RDS survey was designed and conducted to collect demographic, behavioral and serologic data. RDSAT was used to calculate point estimation and confidence intervals. SPSS was used for bi-variate analysis using RDSAT exported weighed data. NETDRAW was used to draw network diagram. Results Among 617 subjects recruited, the adjusted HIV and syphilis prevalence rates were 16.8% and 10.9%, respectively. 73.0% of the subjects were 20 to 29 years old and 72.9% were officially registered residents of Chongqing. 83.4% were single, with the proportion of students the highest, accounting for 24.6%. During the last six months, 83.4% of them reported ever having anal sex, and 54.0% reported having unprotected anal sex. Conclusion This survey confirmed that Chongqing had a higher reported HIV prevalence among MSM than from other Chinese cities. Comprehensive intervention services were required to address this alarmingly high prevalence, with focus on intervention through internet and those having syphilis infection. RDS seemed one of the effective ways of recruiting hidden MSM populations in Chongqing which had a large population of active MSM who did not frequently visit MSM venues as compared with snowball method.