By using cup culture, influence of different strains (ammonium-resistant N2-fixing type, wild type, none N2-fixing type) of Klebsiella oxytoca on the growth of rice seedling was compared. It was discovered that ammonium-resistant N2 -fixing bacteria could excrete some plant growth promoting substance, which could be adsorbed by cation resin. It' s activity wouldn't be affected at 80℃. At optimal concentration, the weight of rice root and seedling were increased by

Based on the physiological properties of ammonium-resistant N2-fixingbacterium ( Klebsiella oxytoca NG13/pMC73A), the fermentation technology of it was studied. The basic medium of high-density culture was established, with glucose as carbon source coupled with appropriate nitrogen source and inorganic salts. At the middle and late phase of culture, glucose and ammonia were added to supply carbon source and nitrogen source, stabilizing the pHat 6.5 ~ 6.8. Optimal level of dissolved oxygen was kept by controlling aeration and stirring rate. Bacterium number of Klebsiella oxytoca NG13/pMC73A reached 600 ~ 700 x 10s cfu/mL at the end of culture. Compared with previous technology, bacterium number was increased by more than ten-fold with a comparable culture period.

BACKGROUND:A steady method that can isolate endothelial progenitor cells(EPCs) need to be explored.Studies show that the EPCs riches in bone marrow with strong reproductive activity,which can be easily obtained.OBJECTIVE:To evaluate the isolating culture and biological characterization of EPCs derived from rat marrow.DESIGN,TIME AND SETTING:The in vitro cytology experiment was performed at the central laboratory of the First Affiliated Hospital of Chongqing Medical University from August to December 2008.MATERIALS:Twenty SD rats,with two-week-old,were obtained from Experimental Animal Center of Chongqing Medical University.METHODS:femurs and tibias of rats were isolated under sterile conditions,and then the bone marrow cavity was washed by 4 ℃ 0.01 mol/L phosphate buffer(PBS).Mononuclear cells were collected from marrow suspension by density gradient centrifugation,followed by cultured with M199 culture medium containing 20% fetal bovine serum to obtained cell suspension.The cells were vaccinated into culture flask(coated with fibronectin) with the density of 1?109/L,and incubated at 37 ℃ in 5% CO2 atmosphere.MAIN OUTCOME MEASURES:Morphologic change of cells was observed by an invert microscope,the expression of CD133,CD34 as well as KDR was detected by immunocytochemistry,and the flow cytometry was used to analyze the growth curve.RESULTS:The adherent cells formed clusters that were found to be characterized by density packed round cells surrounded by spindle-likes cells.Adherent cells were identified positive for CD133,CD34 and KDR by immunocytochemistry on days 4,7,10 and 14.CD133-positive cells reduced after 10 days.Flow cytometry analysis indicated that the number of CD133/KDR double positive cells gradually increased to the maximum at day 10 and then declined.KDR-positive cells gradually increased up with the time.EPCs growth curve showed that cells proliferated fast on days 3-6.CONCLUSION:EPCs can be isolated and cultured by density gradient centrifugation from marrow suspension of SD rats.These cells exhibit the characterization of endothelial cells,and can differentiate into endothelial cells.

Objective To compare the effect of allograft and autograft of bone-patellar tendon- bone (B-PT-B) under arthroscopy in reconstruction of anterior cruciate ligament (ACL).Methods The injured ACL was reconstructed under arthroscopy by using autograft of B-PT-B in 30 cases and al- lograft of B-PT-B in 28 cases from January 2000 to August 2003.After over twelve months follow up,a comparison was done on rehabilitation of quadriceps femoris,knee stability,the Lysholm's score of knee and the enlargement of tunnel in both groups.Results Both groups obtained marked improvement of knee stability and symptoms after operation.But in the early time,the rehabilitation of quadriceps femoris in the ailograft group is faster than that in the autograft group.The rehabilitation of quadriceps femoris, knee stability and the Lysholm's score of knee were similar at 12th month (P＞0.05).There could be seen enlargement of tunnel in both groups,with more in the allograft group (P＞0.05).Conclusion For arthroscopic reconstruction of anterior crucial ligament (ACL),allograft of B-PT-B is good substitute, for it is characterized by effectiveness,convenience and minor side effect.

The ability to induce hypersensity on leaves of tomato and the stability of double-plasmid of an harpin-producing, nitrogen-fixing engineered strain E4 were tested. Hypelsensitivity-inducing experiment indicated that the time and density of hypersensitivity-induction of E4 was similar to those of DH5, the positive control of pCPP430. Although E4 took the same time to induce hypersensitivity as 308R, another positive control of pCPP430, it induced weaker hypersen- sitivity on tobacco leaves. On tomato leaves, there was no difference in time and density of hypersensitivity between E4 and 308R (pCPP430). Results revealed that the two plasmids, pCPP430 and pMC73A, were unstable in host bacteria, with the losing rate of 100% at the 48th generation. The emergence probability of bacteria with either pCPP430 or pMC73A was almost the same.

Objective To investigate the prevalence of subclinical hypothyroidism and its relation to serum total cholesterol (TC),triglyceride (TG) levels and non-alcoholic fatty liver.Methods Serum levels of TC,TG, TSH,TT_3 and TT_4 were determined in 1 602 subjects which were divided into groups by sex and age (20-39,40- 59 and≥60 years).Fatty liver was diagnosed by type B ultrasonography.Restults ( 1 ) Prevalence of subclinical hypothyroidism was 6.6% in this population.(2) The mean levels of serum TC and TG in subclinical hypothyroidism group and normal group showed no significant difference ( both P＞0.05 ).(3) On the whole,the prevalences of subclinical hypothyroidism in hypercholesterolemia group and normal cholesterolemia group showed significant difference (P＜0.01 ).(4) In the three age groups of male and few.ale examinees,the prevalences of subclinical hypothyroidism in hypercholesterulemla group and normal cholesterolemia group showed no significant difference ( all P＞0.05 ).In the multiple regression analysis,TSH level was not related with raised TC level.(5) In the three age groups of male,there was no significant difference between the prevalences of subclinical hypothyroidism in hypertriglyceridemla group and normal triglyceridemia group ( all P＞0.05 ).In female,expect for the elderly group (＞60 years old ),the prevalence of subclinical hypothyroidism was higher in hypertriglyceridemia group than that in normal triglyceridemia.However,in the multiple regression analysis, increased TSH level was positively associated with increased serum TG level ( OR = 1.072,P=0.013 and OR = 1.102,P = 0.03 ).(6) The prevalence of subclinical hypothyroidism in non-alcobolic fatty liver group and normal group showed no significant difference ( P＞0.05 ).In the multiple regression analysis,TSH level was not the independent risk factor of non-alcoholic fatty liver (P＞0.05 ).Conclusion Subclinical hypothyroidism is not asseciated with serum TC level but positively associated with serum TG level.Subclinical hypothyroidism does not apparently increase the prevalence of non-alcoholic fatty liver.

OBJECTIVETo compare the effect of transumbilical single-site single-port with that of transumbilical single-site double-port laparoscopic varicocelectomy in the treatment of varicocele in adolescents.

METHODSWe randomly assigned 80 varicocele patients aged 10 - 16 years to two groups of equal number to receive transumbilical single-site single-port and single-site double-port laparoscopic varicocelectomy, respectively. We compared the operation time, postoperative hospital stay, incisional pain, complications and satisfaction with the abdominal cosmetic outcomes between the two groups.

RESULTSAll the operations were successfully performed. The double-port group showed a significantly higher score on the Visual Analogue Scale than the single-port group (4.8 +/- 1.4 vs 3.6 +/- 1.1, t = -4.986, P < 0.01), but there were no significant differences between the two groups in the operation time ([29.8 +/- 4.2] vs [31.2 +/- 4.6] min, t = 1.383, P = 0.171), postoperative hospital stay ([1.95 +/- 0.7] vs [1.82 +/- 0.8] d, t = -0.784, P = 0.436), complications (0 vs 0) and scores on the satisfaction with abdominal cosmetic outcomes (4.6 +/- 0.6 vs 4.8 +/- 0.5, t = 1.253, P = 0.214). No recurrence, umbilical hernia, hydrocele and orchiatrophy were found in the two groups of patients at 6 months after operation, and no visible scar was observed on the abdominal surface.

CONCLUSIONWith strict surgical indications, single-site single-port and single-site double-port laparoscopic varicocelectomies have similar clinical effects in the treatment of varicocele, which leave no scar on the abdominal surface. Single-site double-port laparoscopy needs no special instruments and therefore is worthier of wide clinical application.

Adolescent ; Child ; Humans ; Laparoscopy ; methods ; Length of Stay ; Male ; Operative Time ; Umbilicus ; surgery ; Varicocele ; surgery

OBJECTIVETo study the effect of acupuncture on the immune function of sepsis patients.

METHODSNinety sepsis patients were assigned to the control group, the thymosin a1 group, and the acupuncture treatment group according to random digit table, 30 patients in each group. Patients in the control group were treated according to the guideline of Surviving Sepsis Campaign (SSC). Patients in the control group received routine treatment. Those in the thymosin alpha1 group additionally received subdermal injection of thymosin alpha1 (1.6 mg), once per day for 6 successive days. Needling at related points such as Zusanli (ST36), Yanglingquan (GB34), Neiguan (PC6), Guanyuan (RN4), and so on, was performed in patients of the acupuncture treatment group, once per day for 6 successive days. T cell subgroups (CD3+, CD4+, CD8+, CD4+ /CD8+) and immunoglobulin levels (IgG, IgA, IgM) were detected. The length of ICU hospital stay, hospital readmission rate, and 28-day mortality were compared among the three groups.

RESULTSAfter six days of treatment, CD3+, CD4+, CD8+, IgG, IgA, IgM, and CD4+ /CD8+ ratio of three groups were all significantly increased (P < 0.01). Of them, CD3+, CD4+, CD8+, IgG, IgA, and IgM increased more significantly in the thymosin alpha1 group and the acupuncture treatment group (P < 0.01). Compared with the control group, the ICU hospitalization length was significantly shortened, the hospital readmission rate and the 28-day mortality were lower in the thymosin alpha1 group and the acupuncture treatment group (P < 0.05, P < 0.01). There was no statistical difference in each index between the thymosin alpha1 group and the acupuncture treatment group (P > 0.05).

CONCLUSIONAcupuncture could adjust the immune function of sepsis patients, improve their immunological indicators and prognoses.

Acupuncture Therapy ; CD4-CD8 Ratio ; Humans ; Length of Stay ; Prognosis ; Sepsis ; diagnosis ; immunology ; therapy ; T-Lymphocyte Subsets ; Thymosin ; analogs & derivatives

Objective To explore the effect and mechanism of relaxin on the production of extracellular matrix (ECM) excreted by high glucose stimulated human renal mesangial cells.Methods Cultured human mesangial cells (HMCs) were divided into three groups:(1) normal glucose group (NG,5.5 mmol/L D-glucose),(2) high glucose group (HG,30 mmol/L D-glucose),and (3) high glucose + relaxin group.Cell count kit (CCK8) was used to examine the cell proliferation.The levels of fibronectin and collagen type Ⅳ in the culture supernatants were examined with a solid-phase enzyme-linked immunoadsorbent assay (ELISA);Western blot method was used to detect the expression of α-smooth muscle actin (α-SMA) protein.The transforming growth factor-β1 (TGF-β1) mRNA expression was detected with quantitative polymerase chain reaction (qPCR) method.Results No proliferation and inhibition effects were observed in both normal and high glucose group.Compared to the normal glucose group,the levels of fibronectin,and collagen type Ⅳ increased significantly (57.28 ± 0.59 vs 41.85 ± 0.03,56.52 ± 0.88 vs 33.80 ± 0.24,P ＜ 0.01)after cultured 48 h in high concentration of glucose.Compared to the high glucose group,a significantly decreases of fibronectin and collagen type Ⅳ (47.08 ± 0.03 vs 57.28 ± 0.59,36.16 ± 0.52 vs 56.52 ±0.88,P ＜0.01) were observed in the relaxin treated group.The expressions of α-smooth muscle actin and TGF-β1 were decreased (P ＜0.01).Conclusions Relaxin can suppress the overproduction of ECM excreted by HMC cultured in high ambient glucose,and its mechanism is partly due to the inhibition of TGF-β1.

OBJECTIVETo observe the clinical effect of acupuncture on acute heart failure (AHF) patients.

METHODSTotally 60 patients who were diagnosed as AHF were assigned to the acupuncture group and the control group, 30 in each group. Those in the control group received inotropic agents, preload and afterload reducing therapy, anti-infection and so on. Besides, those in the acupuncture group received needling at relative points, once daily for 5 consecutive days. The mean arterial pressure (MAP), heart rate (HR), cardiac index (CI), stroke volume index (SI), left ventricle working index (LCWI) were monitored by thermodilution pulse-indicated continuous cardiac output (PiCCO) technique. Changes of the aforesaid data were compared between before and after treatment. The ICU length of stay, readmission rate,and the 28-day mortality were also compared between the two groups.

RESULTSAfter 5 days of the treatment, CI, SI, and LCWI increased more obviously (P < 0.01), HR and MAP decreased significantly (P < 0.05). Of them, CI, SI, and LCWI increased more obviously in the acupuncture group (P < 0.05).There was no obvious difference in HR or MAP between the two groups (P > 0.05). Compared with the first day of admission in the same group, CI, SI, and LCWI obviously increased in the acupuncture group from the second day (P < 0.05). HR started to decrease since the fourth day (P < 0.05), and MAP began to decrease until the fifth day (P < 0.05). CI, SI, and LCWI started to increase in the control group from the third day (P < 0.05); HR and MAP both began to decrease since the fifth day (P < 0.05). Compared with the control group, the ICU length of stay was obviously shortened in the acupuncture group (P < 0.05). The readmission rate and the 28-day mortality rate were lower than those of the control group (P < 0.05).

CONCLUSIONThe combination of acupuncture and Western medical therapy might strengthen acute heart failure patients' heart functions, elevate the therapeutic effect, and improve the prognosis.

Acupuncture Therapy ; Adult ; Aged ; Aged, 80 and over ; Female ; Heart Failure ; physiopathology ; therapy ; Heart Rate ; Humans ; Male ; Middle Aged ; Stroke Volume