OBJECTIVE:To establish a method for simultaneous determination of the contents of ligustroflavone,specnuezhe-nide,demethylwedelolactone and wedelolactone in Zibu ganshen pill. METHODS:Dual-wavelength HPLC was performed on the column of Elite C18 with mobile phase of acetonitrile-0.5%acetic acid(gradient elution)at flow rate of 0.9 ml/min,column temper-ature was 25 ℃,detection wavelengths were 224 nm(0-30 min) and 351 nm (30-50 min),and the injection volume was 20 μl. RESULTS:The linear range was 6.75-135.00μg/ml(r=0.999 5)for ligustroflavone,6.54-130.80μg/ml(r=0.999 8)for specnuezhe-nide,4.90-98.00μg/ml(r=0.999 4)for demethylwedelolactone and 6.42-128.40μg/ml(r=0.999 6)for wedelolactone;RSDs of pre-cision,stability and reproducibility tests were no more than 1.25%;average recoveries were 96.15%-99.96%(RSD<2%,n=6). CONCLUSIONS:The method is rapid,sensitive and accurate,and can be used for the contents determination of ligustroflavone, specnuezhenide,demethylwedelolactone and wedelolactone in Zibu ganshen pill.

Objective To analyse the significance of proto oncogene protein in oral squamous cell carcinoma and oral leukoplakia.Methods 40 patients who were diagnosed with oral leukoplakia (OLK),40 patients who were diagnosed with oral squamous cell carcinoma(OSCC),and at the same time, 40 healthy volunteers as control group were collected.the lesions and normal oral mucosa were taken as samples, left supernatant after grinding, MDM2 protein, p53 protein and Ki-67 protein were detected by ELISA in three groups,and analyse the correlation between three groups.Results Among three groups,MDM2, p53 expression:OSCC group>OLK group>control group (P<0.05).Ki-67 expression level among three groups, OSCC group>OLK group>control group (P<0.05).MDM2, p53, Ki-67 expression had a positive correlation in OSCC group and OLK group (P<0.05), but there was no obvious correlation in control group.Conclusion When oral leukoplakia developes into oral squamous cell carcinoma, MDM2, p53, Ki-67 shows high expression, and there is a positive correlation between MDM2, p53, Ki-67.

Objective: To assess the implementation effects of clinical pathways, compared with usual care, among patients with stroke. Methods:Two investigators independently searched PubMed, Embase, the Cochrane Library, Web of Science, Chinese Biomedical Literature Database and Wanfang Database for studies published before December 2014. Jadad methodological approach was applied to assess the quality of included studies and RevMan software (version 5.2.7) was used for meta-analysis. Results: A total of 11 RCTs involving 913 patients were included in this meta-analysis. The overall results showed that a shorter average length of stay [MD = -2.92; 95% CI (-4.06, -1.78); P < 0.001] and a lower inpatient expenditures [SMD = -1.64; 95% CI (-1.80, -1.48); P < 0.001] in clinical pathways group comparing with the usual care group. The higher score of patient satisfaction was also seen in clinical pathways group. Conclusion: clinical pathways may reduce the average length of stay, reduce the inpatient expenditures, increase patient satisfaction and improve the quality of care in stroke management.

[Objective] Compare the outcomes of vacuum extractors and forceps in assisted vaginal delivery.[Methods] One hundred and sixty-four pregnant women with operative vaginal delivery who received prenatal examination and delivered in our hospital were enrolled.Eighty-two women with vacuum extractors were assigned into vacuum extractors group,another eighty-two women with low or outlet forceps were assigned into forceps group.Occiput transverse or posterior position women before operative vaginal delivery were assigned into abnormal fetal position group.Compare the maternal and neonatal outcomes of vacuum extractors and forceps in assisted vaginal delivery.[Results] The vaginal laceration rate of forceps group was significant higher than vacuum extractors group (56.1％ vs 24.2％,P ＜ 0.01),but neonatal subscalp hematoma and hyperbilirubinemia rate were lower than vacuum extractors group (P ＜ 0.05).The vaginal laceration rate of abnormal fetal position forceps group was also higher than abnormal fetal position vacuum extractors group (76.5％ vs 22.2％,P ＜ 0.05),but neonatal hyperbilirubinemia rate was lower than abnormal fetal position vacuum extractors group (P ＜ 0.05).The Interval time from decision to delivery,postpartum hemorrhage volume with 24 hours,leukocyte and neutrophil count after 24 hours,asphyxia of newborn rate,intracranial hemorrahge,subgaleal hemorrahge,referral rate and hypoxic-ischemic encephalopathy did not significantly differ between vacuum extractors group and forceps group (all P ＞ 0.05).[Conclusions] Vacuum extractors can reduce the incidence of vaginal laceration rate,especial for abnormal fetal position,but increase the incidence of neonatal subscalp hematoma and hyperbilirubinemia rate.

Objective: To assess the implementation effects of clinical pathways, compared with usual care, among patients with stroke. Methods:Two investigators independently searched PubMed, Embase, the Cochrane Library, Web of Science, Chinese Biomedical Literature Database and Wanfang Database for studies published before December 2014. Jadad methodological approach was applied to assess the quality of included studies and RevMan software (version 5.2.7) was used for meta-analysis. Results: A total of 11 RCTs involving 913 patients were included in this meta-analysis. The overall results showed that a shorter average length of stay [MD = -2.92; 95% CI (-4.06, -1.78); P < 0.001] and a lower inpatient expenditures [SMD = -1.64; 95% CI (-1.80, -1.48); P < 0.001] in clinical pathways group comparing with the usual care group. The higher score of patient satisfaction was also seen in clinical pathways group. Conclusion: clinical pathways may reduce the average length of stay, reduce the inpatient expenditures, increase patient satisfaction and improve the quality of care in stroke management.
Stroke

Objective:To develop an HPLC-DAD-ELSD method for the determination of nitidine chloride, 5-ethoxychelerythrine, bergeninum and ardisiacrispin A in Shangtong tinctures ( STD) . Methods: A Hypersil C18 column was used as the chromatographic column, the flow rate was 0. 8 ml·min-1 . For nitidine chloride and 5-ethoxychelerythrine, the mobile phase A consisted of acetoni-trile,the mobile phase B consisted of 0. 1% formic acid-triethylamine (pH 4. 5),and the DAD detection wavelength was at 273 nm. For bergeninum and ardisiacrispin A, the mobile phase consisted of methanol-water(25∶75), the temperature of drift tube was set at 95℃, and the gas flow (N2) was set at 2. 5 SLPM·min-1. Results:There was a good linear relationship between the concentration and peak area for nitidine chloride and 5-ethoxychelerythrine within the range of 0. 021-0. 426 μg (r=0. 999 5) and 0. 075-1. 494 μg (r=0. 999 8), respectively. The average recovery was 99. 22%(RSD=0. 64%) and 98. 61%(RSD=0. 46%), respectively. There was a good linear relationship between the concentration and peak area for bergeninum and ardisiacrispin A within the range of 0. 215-4. 304 μg(r=0. 999 3) and 0. 286-5. 728 μg(r=0. 999 7), respectively. The average recovery was 99. 15%(RSD=0. 77%) and 99. 25%(RSD=0. 56%) accordingly. Conclusion:The method is accurate, sensitive and reproducible, and can be used in the de-termination of nitidine chloride, 5-ethoxychelerythrine, bergeninum and ardisiacrispin A in STD.

Magnetic nano gene vector is one of the non-viral gene vectors, modified by functional group to bind cationic transfect reagents. Coupling magnetofection with the universal lipofection we developed a novel somatic cell transfection method as the so-called liposomal magnetofection (LMF). This approach is potential to provide somatic cell cloning with stable genetic cell lines to cultivate transgenic animals. In order to construct such liposomal magnetic gene vectors complexes system, we used nano magnetic gene vector to combine with liposomal cationic transfect reagents by molecular self-assembly. This vectors system successfully carried exogenous gene and then transfected animal somatic cells. Here, we conducted atomic force microscopy (AFM), zeta potential-diameter analysis and other characterization experiments to investegate the size distribution and morphology of magnetic nanoparticles, the way of the vectors to load and concentrate DNA molecules. Our data reveal that, the LMF of Pig Kidney cells exhibited higher transfection efficiency comparing with the transfection mediated by the commercial lipofectamine2000. Moreover, LMF method overcomes the constraint of transient expression mediated by lipofection. Meanwhile, MTT assay showed low cytotoxicity of LMF. Hence, LMF is a feasible, low cytotoxic and effective method of cell transfection.
Animals ; Cations ; Cell Line ; DNA ; Genetic Vectors ; Kidney ; cytology ; Liposomes ; Magnetics ; Nanoparticles ; Swine ; Transfection

Objective To evaluate the clinical and manometric characteristics of achalasia.Meth-ods Patients diagnosed as having achalasia from July 2010 to July 2014 at our hospital were enrolled.High resolution manometry(HRM)results were analyzed.Data of Eckardt scale,MDADI and SF-36 were ana-lyzed.Results All subjects had dysphagia,35.6% (37 /104)accompanied with regurgitation,26.9%(28 /104)with heartburn and 17.3%(18 /104)with weight loss.According to HRM results and Chicago classification criteria,16.35%(17 /104)of the subjects were classified as type Ⅰ,76.92%(80 /104)as type Ⅱ and 6.73%(7 /104)as type Ⅲ.27.9%(29 /104),19.2%(20 /104)and 24.0%(25 /104)of the subjects finished Eckardt scale,MDADI and SF-36,respectively.Eckardt score was positively correlated with integrated relaxation pressure(IRP)(r =0.421,P <0.05)and MDADI physical score was negatively with IRP(r =-0.530,P <0.05).Conclusion Dysphagia often occurs as the chief complaint among acha-lasia patients.And type Ⅱ is the most common.IRP is an indicator of the severity of clinical symptoms and impairment of quality of life.

Objective:To study the role of phospholipase A 2 in the pathomechanism of low back and leg pain. Methods:Phospholipase A 2 was injected into rats′ cavum epidurale, then the pathologic changes of cauda equina was viewed under light microscopy and electronic microscopy. The substance P(SP) in the dorsal root ganglia(DRG) and spinal cord was determined by immunohistochemistry. Results:After phospholipase A 2 injection, the nerve fibers of cauda equina displayed significant degeneration and demyelination. In L 4 to L 6 segment, the number and area of SP positive neurons in the DRG increased significantly. The relative density of positive nerve endings in the dorsal horn of spinal cord was also increased significantly. There were significant difference compared to the normal solution control group by statistic analysis. Conclusion:Phospholipase A 2 is a nocuous substance to cauda equina. It may play an important role in the pathomechanisms of low back and leg pain related to lumbar disc herniation.

Objective To evaluate safety and clinical significance of single balloon enteroscopy (SBE) for small intestinal diseases. Methods Data of 83 patients with suspected or known small intestinal diseases, who underwent SBE from March 2009 to July 2010, were reviewed in terms of preparation time,procedure time, detection rate and complication occurrence. Results The 83 patients included 37 cases of digestive tract bleeding, 38 chronic abdominal pain, 1 chronic diarrhea, 2 fever and 5 incomplete ileus. A total of 94 procedures of SBE were performed, including oral route in 46 patients, anal route in 26 and both routes in 11. Excluding 6 cases with endoscopic therapy, the mean procedure time of oral approach was 29.6 ± 10. 3 min, and that of anal route was 57.1 ± 15.6 min. Abnormalities were detected in 57 ( 68.7% )of the 83 patients, with detection rate of 81.1% (30/37) in digestive tract bleeding with unknown reason,57. 8% (22/38) in chronic abdominal pain of unknown reason, 50. 0% (1/2) in fever of unknown reason and 80. 0% (4/5) in incomplete ileus. Peutz-Jeghers syndrome was diagnosed in 6 patients and endoscopic polypectomy was performed, with complicated bleeding in one patient. No other procedure-related complications were observed. Conclusion SBE is well-tolerated and safe for diagnosis of small intestine diseases,with easy manipulatiou, short procedure time, high detection rate and satisfactory location of intestinal hemorrhagic lesions.