In this experiment six methods,calcium chloride(CaCl_(2)) precipitation,polyethylene glycol(PEG,pH7.0) precipitation,polyethylene glycol(PEG,pH11.5) precipitation,aluminum chloride(AlCl_(3)) precipitation,Amicon Utcra centrifugal filter devices and cellulose nitrate membrane were used to concentrate the vaccine poliovirus type 1(PV_(1)) added to water body;experimental conditions for concentration were selected and optimized.The results showed that two methods,CaCl_(2)and PEG(pH 7.0) precipitation were suitable for concentrating virus in large volumes of water while amicon utcra centrifugal filter devices for small ones.The virus recovery of the three methods reached a 100% rate.

[ ABSTRACT] AIM:To investigate the effect of phycocyanin on the apoptosis of human laryngeal cancer HEP-2 cells and to explore the inhibitory mechanism of phycocyanin to tumor.METHODS:Highly purified phycocyanin was ex-tracted from spirulina.The effects of phycocyanin at different concentrations on the growth of human laryngeal cancer HEP-2 cells were detected by MTT assay.In addition, the cell structures were observed under electron microscope.The cell ap-optosis was analyzed by flow cytometry.The production of reactive oxygen species ( ROS) was measured by flow cytometry. Enzymatic activities of caspase-3,-8 and-9 were measured by chemical colorimatry.The expression of Bax, Bcl-2, Fas, P53, caspase-3 and caspase-9 at mRNA and protein levels was determined by RT-PCR and Western blot.RESULTS:MTT test confirmed that phycocyanin inhibited the cell activity of HEP-2 cells with time and dose dependent manners.The result of electron microscope observation and flow cytometry indicated that phycocyanin induced the apoptosis of HEP-2 cells.The intracellular content of ROS was increased.The activities of caspase-3, -8 and -9 were increased.RT-PCR showed that the mRNA expression of Bax, Fas, P53, caspase-3, caspase-9 was increased and Bcl-2 was decreased.The results of Western blot were consistent with the results of RT-PCR.CONCLUSION:Phycocyanin might induce apoptosis of HEP-2 cells by down-regulating Bcl-2, up-regulating Bax, Fas and P53, and the transduction of apoptotic signals in the human laryngeal cancer cells.

Bacterial Infections ; diagnosis ; urine ; Colorimetry ; Female ; Humans ; Male ; Middle Aged ; Peritonitis ; diagnosis ; urine ; Urinalysis ; methods

OBJECTIVETo explore the surgical method and clinical curative effect of Ilizarov technique combined with limited surgical treatment for neurotrophic malnutrition equinovarus accompanied with weight-bearing area ulcer.

METHODSFrom July 2004 to December 2011, 21 cases of neurological disorders equinovarus with skin ulcer in weight-bearing area were treated including 14 males and 7 females with an average age of 21.3 years old ranging from 8 to 32 years. Among them,19 cases with talipes equinovarus were on account of spina bifid and 2 cases on account of spinal cord injury of lumbar sacral segment. Nine cases were on the left foot and 12 cases were right foot. The ulcer area in out-below side of the fifth metatarsal bone was in 6 cases, in basement of the fifth metatarsal bone in 5 cases,in lateral of calcaneocuboid joint 2 cases, and in lateral of subtalar joint in 8 cases. Surgical procedure included plantar medial release technique, tendo-chilles lengthenning, and three-joint partial osteotomy. During operation,the skin and soft tissue of ulcer were resected and the incision was sutured when the ulcer was in the lateral of subtalar joint, and ankle Ilizarov external fixation for extension was installed after correcting talipes equinovarus partially. For the legacy skin ulcer in weight-bearing area,the Shenrg-ji cream (Chinese characters) was used after operation for one time per day until the wound healed. Adjusting Ilizarov external fixation for correcting residual deformity until it is satisfaction. During this process weight-bearing walking was keeping.

RESULTSAll cases were followed up from 6 to 52 months with an average of 28 months. All ulcers were healing and no recurrence. The ulcer healing time was 14 to 36 days postoperative (26 days in average). Eighteen deformity feet were corrected satisfactorily, and recovered fully plantar foot, 3 feet appeared mild deformity after 1 year. Ankle joint activity appeared limited in 15 cases and got completely rigidity in 6 cases. The AOFAS increased from preoperative 34.0±7.2 to 86.0±8.5 postoperatively; 8 feet got excellent, 10 good,3 fair.

CONCLUSIONThe clinical effect of Ilizarov technique combined with limited operation and the Sheng-ji cream in correction of the talipes equinovarus with skin ulcer in weigh-bearing area on account of neurotrophic malnutrition is satisfactory, the surgical method is simple and the treating course in security, and serious complications can be avoided.

Adolescent ; Adult ; Child ; Clubfoot ; surgery ; External Fixators ; Female ; Foot Ulcer ; surgery ; Humans ; Ilizarov Technique ; Male

OBJECTIVETo explore the application value of morphology assessment of sperm from fresh semen in routine in vitro fertilization (IVF).

METHODSWe analyzed the morphology of the sperm from fresh or optimized semen samples and, based on the sperm morphology of the raw semen, allocated 908 IVF cycles due to the pure tubal factor to different groups: morphologically normal sperm (MNS) ≤ 4%, > 4% - ≤ 15%, and > 15% in Trial 1 and MNS ≤ 1%, > 1% - ≤ 2%, > 2% - ≤ 3%, and > 3%-- ≤ 4% in Trial 2. We compared the rates of fertilization, cleavage, high-quality embryo, -blastocyst formation, and pregnancy among different groups.

RESULTSThe total fertilization rate was significantly lower in the MNS ≤ 4% than in the MNS > 4% - ≤ 15% and >15% groups (74.40% vs 78.61% and 80.03%, P < 0.01). Compared with the MNS ≤ 1%, > 1% - ≤ 2%, and > 2% - ≤ 3% groups, the MNS > 3% - ≤ 4% group showed remarkably increased rates of 2PN normal fertilization (77.23%, 78.97% and 78.99% vs 85.47%, P < 0.01), cleavage (95.71%, 96.01% and 97.27% vs 98.73%, P < 0.05), and blastocyst formation (53.85%, 49.01% and 49.55% vs 63.41%, P < 0.01). No statistically significant differences were observed in the rates of clinical pregnancy, implantation, early abortion, live birth, or malformation at birth among different groups (P > 0.05).

CONCLUSIONMNS ≤ 4% affected the total rate of fertilization while MNS ≤ 3% reduced the rate of normal fertilization in IVF. However, even MNS ≤ 1% did not result in fertilization disorder or failure. Therefore, teratozoospermia alone was not an indicator of ICSI and sperm mor- phology assessment had no obvious value for predicting the rates of embryo quality, clinical pregnancy, and live birth in IVF.

Female ; Fertilization in Vitro ; Humans ; Male ; Pregnancy ; Pregnancy Outcome ; Spermatozoa ; cytology

Objective To discuss the operative procedures and clinical result of posteromedial and posterolateral approaches in treatment of posterior condylar tibial plateau fractures. Methods From January 2006 to June 2008, 21 patients of posterior condylar tibial plateau fractures were treated by posteromedial and posterolateral knee approaches. There were 12 males and 9 females. The age ranged from 28 to 68 years, with a mean of 39.5 years. Of the patients, 13 had resulted from a traffic accident and 8 had caused by a fall. As for the state of posterior condylar tibial plateau fractures, 7 patients had a medial condylar.fracture, 8 patients had a lateral condylar fracture and 6 patients had a bilateral condylar fracture. Results A follow-up lasted 12-24 months (mean 16.2 months ) in 21 patients. There was no infection, no varus or valgus of the knee, no nerve injuries and loosening or breakage of the screw. All cases had attained bone union. According to the Rasmussen functional scoring, the results were excellent in 12, good in 7 and fair in 2. The excellent and good rate of clinical results was 90.5%. Radiologic results were graded with the Rasmussen score to evaluate the reduction of fracture. There were excellent in 13, good in 7 and fair in 1. The excellent and good rate of clinical results was 95.2%. Conclusion Posteromedial and posterolateral approaches can facilitate the reduction and fixation for posterior condylar tibial plateau fractures. It has many advantages such as good exposure, less invasion and the excellent clinical results.

To study the pharmacokinetics characteristic of loganin, ferulic acid and stilbene glucoside in rat plasma after oral administration of Bushen Tongluo formula. The plasma samples were treated by using liquid-liquid extraction technique, the concentrations were determined by HPLC-UV. Johnson spherigel C18 column (4.6 mm x 250 mm, 5 μm) was adopted and eluted with the of mobile phase of methanol-water containing 0.01% glacial acetic acid in a gradient mode, with the flow rate at 1.0 mL x min(-1), column temperature at 30 degrees C and injection volume of 10 μL. According to the findings, loganin was determined at 235 nm, ferulic acid and stilbene glucoside were determined at 320 nm, with the sample size of 10 μL. The pharmacokinetic parameters of loganin, ferulic acid and stilbene glucoside were calculated by DAS 2. 0 software as follows: C(max) was (0.369 ± 0.042), (0.387 ± 0.071), (0.233 ± 0.044) mg x L(-1); t(max) was (0.226 ± 0.022), (0.282 ± 0.031), (0.233 ± 0.044) h; t(½β) was (6.89 ± 0.20), (10.73 ± 0.11), (6.93 ± 0.09) h; AUC(0-∞) was (1.91 ± 0.36), (3.22 ± 0.52), (1.52 ± 0.33) mg x h x L(-1); AUCO(0-t) was (1.62 ± 0.33), (2.58 ± 0.43), (1.30 ± 0.30) mg x h x L(-1); CL was (20.2 ± 4.0), (1.39 ± 0.23), (31.7 ± 6.9) L x h(-1) x kg(-1), respectively. The results showed that after the oral administration with Bushen Tongluo formula, loganin, ferulic acid and stilbene glucoside showed concentration-time curves in conformity with the two compartment model, with a rapid absorption, loganin and stilbene glucoside was excreted at a moderate speed, and ferulic acid was excreted slowly (but with the highest bioavailability). Bushen Tongluo formula can main maintain plasma concentration with three administrations everyday and so is suitable to be made into common oral preparation.
Administration, Oral ; Animals ; Biological Availability ; Coumaric Acids ; administration & dosage ; blood ; pharmacokinetics ; Drugs, Chinese Herbal ; administration & dosage ; analysis ; pharmacokinetics ; Glucosides ; administration & dosage ; blood ; pharmacokinetics ; Iridoids ; administration & dosage ; blood ; pharmacokinetics ; Male ; Rats ; Rats, Sprague-Dawley ; Stilbenes ; administration & dosage ; blood ; pharmacokinetics

OBJECTIVETo analyze the factors contributing to the occurrence of diabetes insipidus after operations for craniopharyngiomas.

METHODSA total of 121 cases of diabetes insipidus following surgeries for craniopharyngiomas were retrospectively analyzed and the factors associated with postoperative diabetes insipidus were analyzed.

RESULTSThe incidence of diabetes insipidus was 27.3% (33/121 cases) before the operation, 89.9% (107/1119) early after the operation and 39.8%(37/93) in later stages after the operation. The occurrence of early postoperative diabetes insipidus showed a significant relation to the classification and calcification of the craniopharyngioma. Patients with supradiaphragmatic and extraventricular tumors had the lowest incidence of postoperative diabetes insipidus. Late postoperative diabetes insipidus was closely correlated to such factors as age, classification of craniopharyngioma, and intraoperative treatment of the pituitary stalk, but not to the scope of tumor resection or tumor calcification. Late diabetes insipidus was more frequent in children and patients with severed pituitary stalk. The incidence of late postoperative diabetes insipidus was significantly higher in patients with supradiaphragmatic and extra-intraventricular tumors than in those with tumors beneath the diaphragma sellae and extraventricular tumors.

CONCLUSIONSPostoperative diabetes insipidus following surgeries for craniopharyngiomas is closely related to the tumor classification, calcification and pituitary stalk protection.

Adolescent ; Adult ; Aged ; Child ; Child, Preschool ; China ; epidemiology ; Craniopharyngioma ; pathology ; surgery ; Diabetes Insipidus ; epidemiology ; etiology ; Female ; Humans ; Incidence ; Infant ; Male ; Middle Aged ; Neurosurgical Procedures ; adverse effects ; methods ; Pituitary Neoplasms ; pathology ; surgery ; Postoperative Complications ; blood ; etiology ; Regression Analysis ; Retrospective Studies ; Sella Turcica ; Young Adult

Objective Few studies are reported on the correlation of Spinster homolog2(Spns2) with cancer.This study aims to investigate the expression of Spns2 in gastric cancer and the adjacent tissue and explore its effects on the proliferation, clone formation and migration of human gastric cancer cell line SGC-7901.Methods Samples of gastric cancer and the adjacent tissue were collected from 20 patients (12 males and 8 females) after surgery between February 2016 and August 2016. The expression of Spns2 in the gastric cancer and adjacent tissues was determined by immunohistochemistry. Eukaryotic expression vector pEX-1 (pGCMV/MCS/EGFP/Neo)-Spns2 and siRNA were constructed and transfected into the human gastric cancer cell line SGC-7901. The cells were divided into a pEX-1- Spns2 (siRNA-Spns2) group and a negative control pEX-1 (siRNA-NC) group. The mRNA and protein expressions of Spns2 were detected by RT-PCR and Western blot, respectively. And the effects of Spns2 on the migration, clone formation and proliferation of the SGC-7901 cells after Spns2 over-expressed and down-regulated were evaluated by cell counting kit-8 (CCK-8) assay, clone formation assay, transwell experiment and wound healing scratch assay.Results The expression of Spns2 was significantly lower in the gastric cancer than in the adjacent tissue (0.39±0.04 vs 0.45±0.02, P<0.05). After transfection, both Spns2 mRNA and protein levels were remarkably higher in the pEX-1-Spns2 (21.96±2.08 and 10.71±2.78) than in the pEX-1 group (0.88±0.12 and 0.83±0.16) (P<0.05), but markedly lower in the siRNA-Spns2 (0.35±0.07 and 0.53±0.07) than in the siRNA-NC group (0.91±0.09 and 0.92±0.08) (P<0.05). At 48, 72 and 96 hours, the proliferation of the SGC-7901 cells was significantly decreased in the pEX-1-Spns2 group as compared with the pEX-1 group (P<0.05) but increased in the siRNA-Spns2 group in comparison with the siRNA-NC group (P<0.05); the clone formation rate was reduced in the pEX-1-Spns2 group as compared with the pEX-1 group (\[33.30±3.81\]％ vs \[48.00±4.60\]％, P<0.05) but elevated in the siRNA-Spns2 group in comparison with the siRNA-NC group (\[48.38±4.22\]％ vs \[26.25±4.88\]％, P<0.05).Conclusion Spns2 is lowly expressed in gastric cancer, and its over-expression in SGC-7901 cells can reduce its proliferation, clone formation and migration. Spns2 may play a role in inhibiting the development and progression of gastric cancer.

AIMTo investigate the protective effect of lumbrokinase against myocardial ischemia and to further explore its underlying mechanisms.

METHODSThe effect of lumbrokinase on myocardial ischemia was observed by a model of acute myocardial infarction due to permanent ligation of the left anterior descending coronary artery in rats. Patch-clamp technique and laser scanning confocal microscopy were utilized to study the action of lumbrokinase on L-type calcium current (ICa-L) and intracellular calcium concentration ([Ca2+]i).

RESULTSLumbrokinase decreased the infarct size of myocardium in a dose-dependent manner. The inhibitory rate of lumbrokinase at the dose of 20, 40 and 80 mg x kg(-1) was 7.7%, 34.6% and 46.2%, respectively. The electrophysiological studies displayed that, at + 10 mV, the ICa-L was markedly reduced from (-14.42 +/- 1.53) pA/pF to (-11.33 +/- 1.40) pA/pF (decreased by 21.4%, P <0.01) and (-9.92 +/- 1.31) pA/pF (decreased by 36.5%, P <0.01) by lumbrokinase (10 and 50 micromol x L(-1)), respectively. Confocal experiments showed that 10 micromol x L(-1) lumbrokinase showed no obvious effects on [Ca2+]i at resting states (P > 0.05). However, the increase of [Ca2+]i induced by 60 mmol x L(-1) KCl was distinctly limited by 10 micromol x L(-1) lumbrokinase (P <0.01). Within 240 s, the no obvious peak value of fluorescent intensity (FI) was shown.

CONCLUSIONLumbrokinase showed protective action against myocardial infarction in rats. The possible mechanisms of anti-ischemia could be attributed to decreasing ICa-L and [Ca2+] of ventricular myocytes in rats.

Animals ; Calcium ; metabolism ; Calcium Channels, L-Type ; metabolism ; Cardiotonic Agents ; pharmacology ; Endopeptidases ; pharmacology ; Female ; Heart Ventricles ; Male ; Myocardial Infarction ; metabolism ; pathology ; Myocardium ; pathology ; Myocytes, Cardiac ; metabolism ; Rats ; Rats, Wistar