Physiological and biochemical characteristics,drug sensitivity of pathogenic bacteria isolated from the diseased Sparus latus in Zhanjiang,Guangdong,and inactivated whole cell vaccine were studied.The results show that the isolated strain is Vibrio alginolyticus and is highly sensitive to tetracycline,gentamycin,chloramphenicol,and so on.But it is not sensitive to erythromycin and penicillin K.It also shows that the pathogen is inactivated more easily at pH6.0 than at pH8.4 when formalin is used.The whole cell vaccine can strengthen Sparus latus's immuno function and lessen their death rate when fish are infected by the Vibrio alginolyticus.

To clarify the active components in Guizhi Fuling capsule in treatment of intrinsic dysmenorrhea, pelvic inflammation and hysteromyoma, main components were gradually knocked out from the capsules, the effects of knockout capsules on uterine contraction, TNF-α secretion, murine splenocytes (SPL) and hysteromyoma cells proliferation were evaluated, respectively. The inhibition of capsules on uterine contraction was weakened by gradient knockout of paeoniflorin, paeonol, and amygdalin. The suppression of capsulte on TNF-α secretion was reduced by gradient knockout of gallic acid, cinnamaldehyde, pentagalloylglucose, and pachyman. The promotion of SPL cells proliferation was reversed by gradient knockout of gallic acid, paeoniflorin, cinnamaldehyde, quercetin, and pachyman. The depression of capsules on hysteromyoma cells proliferation was attenuated by gradient knockout of paeoniflorin, paeonol, pentagalloylglucose, and albiflorin. In conclusion, the compounds mentioned-above could be the key active basis of Guizhi Fuling capsule in treatment of intrinsic dysmenorrhea, pelvic inflammation and hysteromyoma.
Animals ; Capsules ; administration & dosage ; chemistry ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Dysmenorrhea ; drug therapy ; metabolism ; physiopathology ; Female ; Humans ; Mice ; Mice, Inbred BALB C ; Molecular Imprinting ; methods ; Tumor Necrosis Factor-alpha ; metabolism

Objective To evaluate the effects of fructose-1 ,6-diphosphate (FDP) pretreatment on lung injury induced by hepatic cold liver ischemia-reperfusion in rats .Methods Eighteen healthy male Sprague-Dawley rats were randomly divided into 3 groups (n= 6 each) using a random number table :sham operation group (S group) ,hepatic cold liver ischemia-reperfusion model group (M group) ,and FDP pretreatment group (FP group) . The animals were anesthetized with intraperitoneal chloral hydrate and kept spontaneous breathing .Laparotomy was performed ,and the related blood vessels were only isolated in group S .Hepatic cold ischemia-reperfusion was induced in M and FP groups .In FP group ,FDP 250 mg/kg was injected via the caudal vein at 15 min before skin incision .At 6 h of reperfusion ,the bronchoalveolar lavage fluid (BALF) was collected to detect the levels of tumor necrosis factor-α(TNF-α) ,interleukin-10 (IL-10) and nitric oxide (NO) by ELISA .Lungs were removed for microscopic examination of the pathological changes by light microscopy .Real-time PCR was used to detect the expression of iNOS mRNA .Results Compared with S group , the levels of TNF-α and NO in BALF were significantly increased , the expression of iNOS mRNA was up-regulated , and the level of IL-10 in BALF was decreased in M and FP groups ( P<0.05 ) .Compared with M group ,the levels of TNF-αand NO in BALF were significantly decreased ,the expression of iNOS mRNA was down-regulated ,and the level of IL-10 in BALF was increased in FP group ( P<0.05 ) .The pathological changes of lungs were significantly attenuated in FP group as compared with M group .Conclusion FDP pretreatment can obviously attenuate lung injury induced by hepatic cold ischemia-reperfusion in rats ,and inhibition of iNOS expression ,reduction of NO synthesis ,and decrease in inflammatory responses are involved in the mechanism .

Limit test of flavones in diterpene ginkgolides meglumine injection materials by UV-Vis and HPLC-DAD method was studied in this essay. The HPLC-DAD method has lower LOD (about 1% of the UV-Vis), that is, the sensitivity is higher than UV-Vis method. Through the analysis of the kinds of flavonoids ingredients in the samples by LC-MS, the three compounds with highest contents are kaempferol, quercetin and isorhamnetin. Kaempferol, quercetin and isorhamnetin were chosen as reference compounds for HPLC analysis, and the HPLC separation analysis was carried on an Agilent Eclipse plus C18 column (4.6 mm x 250 mm, 5 μm) with methanol and water containing 0.4% phosphoric acid (50: 50) as mobile phase, and the flow rate was 1.0 mL x min(-1). The detection wavelength was set at 360 nm. This method has good specificity, precision and reproducibility. The LODs of quercetin, kaempferide and isorhamnetin were 27.6, 22.3, 29.5 μg x L(-1). The average recovery was 87.9% (RSD 3.3%), 91.7% (RSD 3.1%), 88.3 (RSD 1.3%) for quercetin, kaempferide and isorhamnetin, respectively. Based on the 10 batches of sample results and sensitivity of different HPLC, the content of total flavonoids ingredients of diterpene ginkgolides meglumine injection materials was limited no more than 2 x 10(-5). This method is simple, quick and has good maneuverability, and could be used to the limit test of flavonoids in the diterpene ginkgolides meglumine injection materials.
Chromatography, High Pressure Liquid ; methods ; Diterpenes ; analysis ; Drugs, Chinese Herbal ; analysis ; Flavones ; analysis ; Ginkgolides ; analysis ; Limit of Detection ; Mass Spectrometry ; methods

Objective To study the relationship between plasma TGF-β, TNF-α, IL-10 levels and radiation pneumonitis (RP) in patients received thoracic irradiation with 3DCRT. Methods Sixty-nine patients of lung cancer stage Ⅲ or esophageal carcinoma were evaluated prospectively by EUSA for plasma TNF-α, TGF- β, IL-10 levels and IL-10/TNF-α before 3DCRT, after 40 - 50 Gy and after 3DCRT. Results Twenty-eight patients had RP. In RP patients, the plasma TGF-β, TNF-α, IL-10 levels and IL-10/TNF-α was (15.2 ± 13.4) μg/L, (28.4 ± 13.4), (24. 1 ± 17. 1) ng/L and 1.01 ± 0.86 before 3DCRT, respectively;TNF-α increased to (36.1 ± 15.5) ng/L(t = 2.01, P = 0.040), IL-10 and IL-10/TNF-α decreased to (18.8 ± 10.8) ng/L (t =1.40, P = 0.166) and 0.62 0.55 (t = 1.90, P = 0.063)after 40-50 Gy. After 3DCRT TNF-α was higher (36.9 ± 15.5) ng/L than that before 3DCRT(t = - 2.20, P = 0.032) ,but IL-10 and IL-10/TNF-α were lower than that before 3DCRT [(13.7 ± 6.2) ng/L, t = 3.03, P = 0.005 ;0.41 ± 0.21, t = 3.60, P = 0.001]. TGF-β was not change in three times(P > 0.05) .In non-RP patients, TGF-β,TNF-α, IL-10 and IL-10/TNF-α was not yet change in three times(P > 0.05) respectively. TGF-β was not yet change between RP and non-RP patients before 3DCRT (t = 0.54, P = 0.594), and TNF-α was higher in RP group than that in non-RP group after 40-50 Gy(t = 2.02, P = 0.048), but IL-10 and IL-10/TNF-α was less in RP group than that in non-RP group after 3DCRT(t=2.50,P=0.015;t=4.63,P=0.000). Conclusions The levels of TNF-α and IL-10 are closely related to the occurrence of RP. Monitoring the changes in dynamic state could predict the generation of RP, which could be employed as a sensitive index for indicating risks for acute RP.

To explore the safety and efficacy of left atrial appendage (LAA) occlusion under the guidance of local anesthesia and modified transseptal puncture technology by coronary sinus assisted positioning in patients with nonvavular atrial fibrillation (NVAF). Methods: A total of 16 NVAF patients received local anesthesia and percutaneous LAmbre or Amplatzer cardiac plug occluder implantation. There were 12 males and the patients mean age was at (71.0±6.0) years with CHA2DS2-VASc score at (4.1±1.5); all patients had walfarin contradiction or with walfarin related side effect. Transseptal puncture was conducted by coronary sinus catheter as the anatomic location marker. Results: All 16 patients finished transseptal puncture and no relevant complication occurred. 15/16 (93.8%) patients had successful LAA occlusion, 1 patient was abandoned because of LAA anatomic structure variation. The mean operative time was (65.0±23.0) min and the mean X-ray exposure time was (12.0±3.0) min. The mean diameter of occluder was (32.5±6.0)mm. Conclusion: LAA occlusion was safe and effective with local anesthesia and modified transseptal puncture technology by coronary sinus assisted positioning in relevant patients.

Objective To explore the incidence and risk factors of acute graft-versus-host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Methods The clinical data of 72 cases allo-HSCT from Oct 2004 to Dec 2008 were analyzed. Thirteen factors possibly correlated with the development of aGVHD were analyzed. Results aGVHD was developed in 32 cases (44.4 %), in which grades Ⅰ aGVHD was 11.1%, gradesⅡaGVHD was 18.1%, and grades Ⅲ-Ⅳ aGVHD was 15.3 %. The univariate analysis showed that diagnosis, the status of disease, use ATG, conditioning regimen, donor type,ABO blood group disparity between donor and recipient, CD34+ cell number, early engraftment and neutropenic infection, HLA locus were associated with the occurence of aGVHD (P ＜0.1). On the COX regression mode, an increased risk of aGVHD was associated with HLA mismatch (HR =2.58, P ＜0.005), GVHD prophylaxis without ATG (HR =2.94, P ＜ 0.001), and unrelated donor (HR =1.97, P ＜0.01). Conclusion aGVHD is a common complication after allo-HSCT, and HLA mismatch and unrelated donor are independent risk factors for aGVHD.

Objective To study the effects of endothelial nitric oxide synthase (eNOS) gene transfection on endothelial progenitor cells (EPCs) transplantation in the process of injured vascular endothelium repair. Methods EPCs were cultured and expanded in vitro. EPCs were transduced with pseudotyped retroviral vectors expressing eNOS gene (pMCV-eNOS-EPCs) or green fluorescent protein gene (pMCV-GFP-EPCs). EPCs with expressing eNOS, GFP or saline were injected respectively into rat injured artery model by tail vein injection after balloon injury and again 24 hours. 14 days after transplantation. eNOS expression in injured artery was detected by RT-PCR, western blot and immunohistochemical methods. The morphology of arterial intima and media was studied by optical microscopy and image analysis system. Results Compared with GFP-EPCs group and control group, the mRNA and protein of eNOS were obviously high expressed in eNOS-EPCs group. EPCs transplantation reduce lumen stenosis and inhibit neointimalhyperplasia (eNOS-EPCs group vs.control group, 0.58±0.05 vs. 1.56±0.21, P < 0.01;GFP-EPCs group vs. control group, 0.84±0.09 vs.1.56±0.21, P < 0.05). eNOS gene transfection could further enhance this anti-proliferative effects (eNOS-EPCs group vs. GFP-EPCsgroup,0.58±0.05 vs. 0.84±0.09, P < 0.05). Furthermore, eNOS modified EPCs could improve the endothelial function of injured vascular endothelium. Conclusions eNOS gene transfection could increase the anti-proliferative effect of EPCs transplantation on injured artery and obviously ameliorate endothelial function.

Taking guizhi fuling capsule (GZFL) for instance, a new method about reference Chinese medicine preparation which was used as standard substance for the quality evaluation of complex Chinese medicine preparation by the fingerprint of reference preparation instead of standard fingerprint was proposed. It could eliminate the errors from different instruments, chromatographic columns and solve the problem of similarity matching in the absence of standard fingerprint. The qualification of reference GZFL was evaluated according to the quality control method of GZFL from Chinese Pharmacopoeia. Then multiple batches of GZFL were estimated, taking fingerprint of reference preparation and standard fingerprint as references, respectively, at different instruments and chromatographic columns. Finally, the packaging and expiration date for reference GZFL were confirmed according to the results of stability investigation. The results indicated that the fingerprint of reference GZFL could be used to assess the quality of GZFL better than standard fingerprint. The data of accelerated stability and long-term stability test demonstrated that reference GZFL was stable in the conditions of double blister package. Therefore, reference GZFL can be used as standard substance in quality control of GZFL.
Capsules ; Drug Stability ; Drugs, Chinese Herbal ; chemistry ; standards ; Reference Standards

The identity of higher-order neurons and circuits playing an associative role to control renal function is notwell understood.We identified specific neural populations of rostral elements of brain regions that project multisynaptically to the kidneys in 3~ days after injecting a retrograde tracer pseudorabies virus (PRV)-614 into kidney of 13 adult male C57BL/6J strain mice.PRV-614 infected neurons were detected in a number of mesencephalic (e.g.central amygdala nucleus),telencephalic regions and motor cortex.These divisions included the preoptic area (POA),dorsomedial hypothalamus (DMH),lateral hypothalamus,arcuate nucleus (Arc),suprachiasmatic nucleus (SCN),periventricular hypothalamus (PeH),and rostral and caudal subdivision of the paraventricular nucleus of the hypothalamus (PVN).PRV-614/Tyrosine hydroxylase (TH) double-labeled cells were found within DMH,Arc,SCN,PeH,PVN,the anterodorsal and medial POA.A subset of neurons in PVN that participated in regulating sympathetic outflow to kidney was catecholaminergic or serotonergic.PRV-614 infected neurons within the PVN also contained arginine vasopressin or oxytocin.These data demonstrate the rostral elements of brain innervate the kidney by the neuroanatomical circuitry.