OBJECTIVETo study the effect of Zhibai Dihuang Pill (ZBDHP) on urokinase-type plasminogen activator (uPA) and sperm quality in ureaplasma urealyticum (Uu) infection infertile patients.

METHODSRecruited were 80 infertility patients with Uu infection at Andriatrics Clinics and Department of Reproduction, including 130 cases of positive Uu semen and 50 cases of negative Uu semen. Patients with positive Uu semen were randomly assigned to the observation group (72 cases) and the control group (58 cases) according to the visit sequence. All patients took antibiotics for 2 weeks. Patients in the observation group additionally took ZBDHP, 6 g each time, twice daily. Those in the control group additionally took Vit E (100 mg each time, twice per day) and ATP (40 mg each time, twice per day). The therapeutic course for all was 90 days. Semen parameters and uPA contents of the sperm membrane were detected and comparatively analyzed.

RESULTSThe sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm in Uu positive infected patients were lower than those in Uu negative infected patients with statistical difference (P < 0.05), but with no significant difference in the sperm density between the two groups (P > 0.05). There was no statistical difference in pre-treatment sperm membrane uPA contents and sperm parameters between the two groups (P > 0.05). Compared with before treatment in the same group, the sperm membrane uPA content, the sperm motility, the sperm viability, and the percentage of normal morphology sperm obviously increased in the two groups with statistical difference (P < 0.05). After treatment, the sperm membrane uPA content increased more obviously in the observation group, with statistical difference when compared with the control group (P < 0.05).

CONCLUSIONSInfection with Uu leads to decreased uPA content of sperm membrance and the sperm motility. ZBDHP could effectively treat Uu infected infertility possibly through fighting against Uu damaged sperm membrane and make the sperm membrane uPA content return to normal, and elevate the fertilizability of sperms.

Anti-Bacterial Agents ; administration & dosage ; pharmacology ; therapeutic use ; Communicable Diseases ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; therapeutic use ; Humans ; Infertility ; Infertility, Male ; Male ; Semen ; Semen Analysis ; Sperm Count ; Sperm Motility ; Spermatozoa ; Ureaplasma Infections ; drug therapy ; Ureaplasma urealyticum ; drug effects ; Urokinase-Type Plasminogen Activator ; metabolism

To evaluate the visual performance of the patients with traumatic corneal astigmatism, after the treatment of topography guided off-flap epipolis laser in situ keratomileusi (off-flap Epi-LASlK).
●METHODS: This prospective clinical study was comprised of 21 eyes of 21 patients with irregular corneal astigmatism caused by trauma, they were treated by off-flap Epi - LASlK from July 2012 to December 2013. The data included uncorrected visual acuity ( UCVA), best spectacle - corrected visual acuity ( BSCVA ), contrast sensitivity 1, 6mo before and after surgery; the healing area percentage of corneal epithelia, the healing time of corneal epithelia and pain score at 3d after surgery.
●RESULTS: Postoperative 1mo both UCVA and BSCVA were improved significantly than that before surgery (t =15. 703, 4. 351, P< 0. 05); Compared with the 1mo after surgery, UCVA at 6mo after surgery raised significantly (t= 6. 867, P <0. 05). There was no statistical significance between 6 and 1mo after surgery about BSCVA (t= 1. 497, P = 0. 140 ). After surgery, mean spherical equivalent (SE) was reduced from -2. 43±3. 02D to -0. 23±0. 49D (P<0. 05), and the mean cylinder was reduced from -1. 86± 2. 23D to - 0. 46 ± 1. 03D (P< 0. 05). Postoperative 1mo,4 kinds of spatial frequency and contrast sensitivity had no significant difference compared with the preoperative (P>0. 05 ). Postoperative 6mo except the 3c/ d spatial frequency, the remaining 3 spatial frequency contrast sensitivity compared with those before operation were significantly improved ( P < 0. 05 ). The healing area percentage of corneal epithelia was 92. 46% ±8. 24% (80% -100%) at 3d after surgery; The healing time of corneal epithelia was 3. 50 ± 1. 56d; Pain scores at 3 and 7d after surgery was 1. 54±1. 32 and 0. 04±0. 64, respectively.
●CONCLUSlON: Topography-guided off-flap Epi-LASlK is safe and effective in treating the patients with traumatic corneal irregular astigmatism. The operation can improve both the contrast sensitivity and the visual performance.

To develop an analytic method for qualitative and quantitative analysis of dodecatetraenamides A, B in 42 samples of two official species of Asari Radix et Rhizoma( ARR) (37 samples of Asarum heterotropoides var. mandshuricum with different collection time and 5 samples of Asarum sieboldiivar. seoulense). The HPLC-IT-TOF-MS/MS methods for the qualitative and UPLC-PDA methods for the quantitative analysis were established. Dodecatetraenamides A, B were identified by comparing the retention time, UV absorption spectrum and quasi-molecular ion peak [ M + H]+ with the reference compound using HPLC-IT-TOF-MS/MS. The content of dodecatetraenamides A and B in ARR were determined by UPLC-PDA. The separation was successfully carried out on a ACQUITY UPLC BEH C18 (2.1 mm x 100 mm, 1.7 µm) column eluted with mobile phases of water (A) and acetonitrile (B) in gradient program (0-3 min, 35% B; 3-5 min, 35%-36% B; 5-6 min, 36%-43% B; 6 min-11 min 43% B; 11-12 min, 43%-100% B). The column temperature was 45 °C, and the detection wavelength was set at 254 nm. The flow rate was 0.6 mL · min(-1). On one level mass spectrometry scanning, the results showed that the quasi-molecular ion [M + H] + of both dodecatetraenamides A and B were m/z 248.20. The quantitative method with UPLC-PDA has made the baseline separation of the constituents, which were reported as mixtures in the most literatures. The average recovery of dodecatetraenamides A and B were 97.90% and 99.86%, the relative standard deviation were 0.4% and 1.1%, respectively. The contents of dodecatetraenamides A, B in all ARR samples was in the range of 0.11-3.89 and 0.24-6.65 mg · g(-1). Their contents reduced with the extension of storage time. Compared with the samples of 2013, the average content of the two constituents in the samples collected in year 2002-2003 reduced 34% and 36%, respectively (P < 0.05). Compared the A. sieboldii var. seoulense and A. heterotropoides var. mandshuricum with the same collective time and production area, the average contents of the two constituents in latter were up to (1.59 ± 0.75) mg · g(-1) and (2.90 ± 1.17) mg · g(-1), respectively, significantly higher than that in A. sieboldii var. seoulense (dodecatetraenamide A were (0.78 ± 0.52) mg · g(-1), dodecatetraenamide B were (1.69 ± 0.83) mg · g(-1)) (P < 0.05). The content of the dodecatetraenamide A in overground part was in the range of 0.11-0.33 mg · g(-1), dodecatetraenamide B was 0. 24-0.60 mg · g(-1), which were much lower than that of the underground part of ARR (dodecatetraenamide A was in the range of 0.73-3.89 mg · g(-1), dodecatetraenamide B was 2.11-6.24 mg · g(-1)). The method was certified to be simple, accurate and reliable and could be used for qualitative and quantitative analysis of dodecatetraenamide A and B in different species of ARR, also can be used for the comprehensive quality control of traditional Chinese medicine, Asari Radix et Rhizoma.
Amides ; chemistry ; Asarum ; chemistry ; Chromatography, High Pressure Liquid ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Mass Spectrometry ; Molecular Structure ; Rhizome ; chemistry

OBJECTIVETo observe the changes in the activity of dendritic cells (DCs) after carcino-embryonic antigen (CEA) gene transfection mediated by recombinant adeno-associated virus type2 (rAAV) and tumor cell lysate.

METHODSImmature DCs isolated from peripheral blood monocytes of HLA-A11-positive healthy volunteers were infected with the rAAV carrying CEA gene or loaded with tumor cell lysate. The surface markers of the DCs such as CD40, CD 1alpha, and CD86 were analyzed by flow cytometry. Interleukin-12 (IL-12) in the supernatants of DCs and interferon-gamma (IFN-gamma) released by the cytotoxic T lymphocytes (CTLs) were determined by ELISA detection kit. The specific killing activity of CTL against LoVo cells was assessed by MTT assay.

RESULTSThe DCs following antigen loading with the two methods both highly expressed CD40, CD86 and IL-12, and induced specific CTL that specifically recognized and killed LoVo cells, but the killing effect resulting from rAAV infection of the DCs was much better than that induced by tumor cell lysate loading.

CONCLUSIONBoth methods of antigen loading can induce mature DCs from peripheral blood monocyte cells, but rAAV infection of the DCs can be more effective than tumor cells lysate loading. DCs infected with rAAV may have the potential to serve as an adjuvant immunotherapy for patients with colorectal carcinoma.

B7-2 Antigen ; metabolism ; CD40 Antigens ; metabolism ; Cancer Vaccines ; biosynthesis ; immunology ; Carcinoembryonic Antigen ; genetics ; Cell Line, Tumor ; Colorectal Neoplasms ; therapy ; Dendritic Cells ; immunology ; metabolism ; Dependovirus ; genetics ; Genetic Vectors ; Humans ; Interleukin-12 ; metabolism ; Transfection

OBJECTIVETo observe the effects of Stragalus membranaceus injection on nitric oxide and endothelin levels of intestinal mucosa in reperfusion injury after hemorrhage shock.

METHOD32 SD rats were randomly divided into four groups: normal group, model group, low dosage group, (treated with Astragalus membranaceus 10 g x kg(-1)); high dosage group (treated with Astragalus membranaceus 20 g x kg(-1)). Models of hemorrhagic shock for 60 minutes and reperfusion for 90 minutes were created. The animals were administrated 3 mL therapeutic solution before reperfusion. At the end of study, intestinal pathology was observed, and the concentration of lactic acid (LD), nitric oxide (NO), endothelin (ET) of intestinal mucosa were detected.

RESULTThe intestinal pathology showed that intestinal mucosa epithelial cells damage in model group was severe, in low dosage group was medium, in high dosage group was slight, and no obvious damage was found in normal group. The concentration of LD and NO of small intestine mucous membrane in model group and low dosage group were significantly higher than those in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05). The concentration of ET of small intestine mucous membrane in model group was the highest of the four groups (P < 0.05). The concentration of ET in low dosage group was significantly higher than that in high dosage group and normal group (P < 0.05), but there were no significant differences between high dosage group and normal group (P > 0.05).

CONCLUSIONStragalus membranaceus injection can reduce small intestine mucous damage by protecting endothelium function in injury after hemorrhage shock-reperfusion.

Animals ; Astragalus membranaceus ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; isolation & purification ; pharmacology ; Endothelins ; metabolism ; Ileum ; metabolism ; pathology ; Injections, Intravenous ; Intestinal Mucosa ; metabolism ; pathology ; Lactic Acid ; metabolism ; Male ; Nitric Oxide ; metabolism ; Plants, Medicinal ; chemistry ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; etiology ; metabolism ; pathology ; Shock, Hemorrhagic ; complications

Objective To investigate the effects of Panax Notoginseng saponins (PNS) on protein expression of Klotho in rats with renal ischemia reperfusion injury; To discuss its protective mechanism for model rats. Methods Experimental rats were randomly divided into sham-operation group, model group, positive medicine group, PNS high-, medium- and low-dosage groups. Each administration group was given relevant medicine for gavage, once a day. Renal ischemia reperfusion injury model was established. Rats were sacrificed by taking blood from abdominal aorta after 4 hours of modeling. Serum levels of blood urea nitrogen (BUN), creatinine (SCr), malondialdehyde (MDA) content in kidney tissue, superoxide dismutase (SOD) activity and glutathione peroxidase (GSH-Px) activity were measured. HE staining was used to observe the morphological changes of renal tissue. The protein expressions of Klotho and NF-κB p65 were measured by immunohistochemical method. Results Compared with the sham-operation group, the levels of BUN and SCr in the model group increased significantly (P<0.05); protein expression of Klotho in renal tissue decreased and the protein expression of NF-κB p65 increased (P<0.05). Compared with the model group, the expression of Klotho increased but protein expression of NF-κB p65 decreased in each administration group (P<0.05); Compared with the positive medicine group, the expression of Klotho in PNS high-dosage group increased but protein expression of NF-κB p65 decreased (P<0.05). The protein expression of NF-κB p65 was negatively related to protein expression of Klotho (r=-0.895, P<0.05). Conclusion PNS can inhibit oxidative stress and anti-inflammatory effects through upregulating protein expression of Klotho, and reduce the protein expression of NF-κB p65, and thus exerts renal protective effects.

Background Noninvasive positive pressure ventilation (NIPPV) has been proposed to shorten the duration of mechanical ventilation in intubated patients,especially those who fail initial weaning from invasive mechanical ventilation (IMV).However,there are also some discrepancies in terms of weaning success or failure,incidence of re-intubation,complications observed during study and patient outcomes.The primary objective of this update was to specifically investigate the role of NIPPV on facilitating weaning and avoiding re-intubation in patients intubated for different etiologies of acute respiratory failure,by comparing with conventional invasive weaning approach.Methods We searched randomized controlled trials (RCTs) comparing noninvasive weaning of early extubation and immediate application of NIPPV with invasive weaning in intubated patients from PubMed,Embase,Cochrane Central Register of Controlled Trials,Web of Knowledge and Springerlink databases.Records from conference proceedings and reference lists of relevant studies were also identified.Results A total of 11 RCTs with 623 patients were available for the present analysis.Compared with IMV,NIPPV significantly increased weaning success rates (odds ratio (OR):2.50,95％ confidence interval (C/):1.46-4.30,P=0.0009),decreased mortality (OR:0.39,95％ CI:0.20-0.75,P=0.005),and reduced the incidence of ventilator associated pneumonia (VAP) (OR:0.17,95％ CI:0.08-0.37,P ＜0.00001) and complications (OR:0.22,95％ CI:0.07-0.72,P=0.01).However,effect of NIPPV on re-intubation did not reach statistical difference (OR:0.61,95％ CI:0.33-1.11,P=0.11).Conclusions Early extubation and immediate application of NIPPV is superior to conventional invasive weaning approach in increasing weaning success rates,decreasing the risk of mortality and reducing the incidence of VAP and complications,in patients who need weaning from IMV.However,it should be applied with caution,as there is insufficient beneficial evidence to definitely recommend it in terms of avoiding re-intubation.

BACKGROUNDCervical keratinocytes are recovered at a low numbers and frequently associated with contaminating human fibroblasts which rapidly overgrow the epithelial cells in culture with medium supplemented with 10% fetal bovine serum (FBS). However, it is difficult to initiate keratinocyte cultures with serum-free keratinocyte growth medium alone because cell attachment can be poor. Therefore, the culture of these cells is extremely difficult. In this study, we described a modified culture medium and coated culture plastics for growing normal human cervical epithelial cells in vitro.

METHODSNormal cervical epithelial tissue pieces were obtained and digested with type I collagenase to dissociate the cells and a single cell suspension produced. The cells were cultured on plastic tissue culture substrate alone or substrate coated with collagen type I from rat tail, with modified keratinocyte serum-free medium (K-SFM) supplemented with 5% FBS. After attachment, the medium were replaced with K-SFM without FBS. The expression of basal keratins of the ectocervical epithelium, K5, K14 and K19 were assayed by immunofluorescence with monoclonal antibodies to identify the cell purity.

RESULTSOur results indicate that cells attached to the culture plastic more quickly in K-SFM supplemented with 5% FBS than in K-SFM alone, as well as to tissue culture plastic coated with collagen type I than plastic alone. The modified medium composed of K-SFM and 5% FBS combined with a specific tissue culture plastic coated with collagen type I from rat tail was the best method for culture of normal cervical epithelial cells. K5, K14 and K19 were assayed and keratinocyte purity was nearly 100%.

CONCLUSIONA novel, simple and effective method can be used to rapidly obtain highly purified keratinocytes from normal human cervical epithelium.

Cell Culture Techniques ; methods ; Cervix Uteri ; cytology ; Epithelial Cells ; cytology ; Female ; Humans ; Keratinocytes ; cytology

BACKGROUNDNoninvasive positive pressure ventilation (NIPPV) has been proposed to shorten the duration of mechanical ventilation in intubated patients, especially those who fail initial weaning from invasive mechanical ventilation (IMV). However, there are also some discrepancies in terms of weaning success or failure, incidence of re-intubation, complications observed during study and patient outcomes. The primary objective of this update was to specifically investigate the role of NIPPV on facilitating weaning and avoiding re-intubation in patients intubated for different etiologies of acute respiratory failure, by comparing with conventional invasive weaning approach.

METHODSWe searched randomized controlled trials (RCTs) comparing noninvasive weaning of early extubation and immediate application of NIPPV with invasive weaning in intubated patients from PubMed, Embase, Cochrane Central Register of Controlled Trials, Web of Knowledge and Springerlink databases. Records from conference proceedings and reference lists of relevant studies were also identified.

RESULTSA total of 11 RCTs with 623 patients were available for the present analysis. Compared with IMV, NIPPV significantly increased weaning success rates (odds ratio (OR): 2.50, 95% confidence interval (CI): 1.46 - 4.30, P = 0.0009), decreased mortality (OR: 0.39, 95%CI: 0.20 - 0.75, P = 0.005), and reduced the incidence of ventilator associated pneumonia (VAP) (OR: 0.17, 95%CI: 0.08 - 0.37, P < 0.00001) and complications (OR: 0.22, 95%CI: 0.07 - 0.72, P = 0.01). However, effect of NIPPV on re-intubation did not reach statistical difference (OR: 0.61, 95%CI: 0.33 - 1.11, P = 0.11).

CONCLUSIONSEarly extubation and immediate application of NIPPV is superior to conventional invasive weaning approach in increasing weaning success rates, decreasing the risk of mortality and reducing the incidence of VAP and complications, in patients who need weaning from IMV. However, it should be applied with caution, as there is insufficient beneficial evidence to definitely recommend it in terms of avoiding re-intubation.

Humans ; Randomized Controlled Trials as Topic ; Respiration, Artificial ; methods ; Ventilator Weaning ; methods