Objective To evaluate the development of immature oocytes after freezing-thawing by conventional cryopreservation method for mature oocytes.Methods Immature oocytes were collected from stimulated ovaries of intracytoplasmic sperm injection(ICSI)cycles.Immature oocytes were in vitro matured directly or after slow freezing-fast thawing and immunostained for tubulin and chromatin and at last visualized by confocal microscopy.Results No statistical difference was found in maturity rate between freezing groups and the controls.There was a statistically significant increase in abnormalities of chromosome(23.7% vs. 50%)and spindle(28.9% vs.53.9%)in the GV freezing group compared with the GV control(P

OBJECTIVETo observe the effect of Kanli Granule (KG) on myocardial mechanics in pressure overload induced diastolic heart failure (DHF) rats.

METHODSTotally 60 male Wistar rats were divided into the sham-operation group, the model group, the KG group, and the Valsartan group according to random digit table, 15 in each group. The pressure overload induced DHF model was established in all groups except the sham-operation group using abdominal aortic constriction surgery. Totally 7 rats died after modeling (with the mortality of 10. 67%) , and the rest 53 finished the following test. Rats in the KG group were administered with KG extract (calculated as 6. 75 g crude drug/kg) by gastrogavage. Rats in the Valsartan group were administered with Valsartan (7.2 µg/g) by gastrogavage. Equal volume of double distilled water was administered to rats in the model group and the sham-operation group by gastrogavage. All rats were intervened for 32 weeks. The response of isolated heart papillary muscle tonus to isoprenaline (ISO) and adenylate cyclase (Forskolin) was respectively observed. The enhancement phenomenon after resting development force (DF) of isolated heart papillary muscle tonus, and changes of DF in different Ca²⁺ concentrations were observed.

RESULTS(1) In the ISO response test: Compared with the sham-operation group, the amplifications of DF, ±df/dt, -df/dt were obviously elevated in the model group (P < 0.05). Compared with the model group, the amplifications of DF and ±df/dt were obviously lowered in the KG group (P < 0.01), and the amplification of ±df/dt was also reduced in the Valsartan group (P < 0.01). (2) In the Forskolin response test: Compared with the sham-operation group, the amplifications of DF and ±df/dt obviously increased in the model group (P < 0.05). Compared with the model group, the amplifications of DF and ±df/dt were obviously reduced in the KG group (P < 0.01), and the amplification of DF was also reduced in the Valsartan group (P < 0.05). (3) In post-resting DF enhancement test: Compared with the sham-operation group, the amplification of DF showed gradually decreasing tendency along with prolonged resting time in the model group, and they were obviously lowered at all time points (P < 0.05). Compared with the model group, the amplification of DF was gradually increasing along with prolonged resting time in the KG group. The amplification of DF at post-resting 240 s was obviously larger in the KG group than in the model group (P < 0.05). The amplification of post-resting DF still showed gradually decreasing tendency along with prolonged resting time in the Valsartan group, with increased amplifications of DF at post-resting 60 s and 120 s (P < 0. 05) (4) The amplifications of DF in different Ca²⁺ concentrations: Compared with the sham-operation group, the amplifications of DF were significantly elevated in different Ca²⁺ concentrations (1.75, 3.5, 7.0 mmol/L ) (P < 0.05, P < 0.01). Compared with the model group, there was no statistical difference in amplification of DF in different Ca²⁺ concentrations in the KG group (P > 0.05). The amplifications of DF in different Ca²⁺ concentrations were significantly reduced in the Valsartan group (P < 0.05).

CONCLUSIONSThe ISO response and the Forskolin response were enhanced in isolated heart papillary muscle tonus of pressure overload induced DHF rats; enhanced post-resting DF was reduced; DF in different supra-physiologic levels of Ca²⁺ was still enhanced. KG could significantly improve excessive enhancement of pressure overload induced DHF rats in ISO response and Forskolin response, and improve enhancement of post-resting myocardium.

Animals ; Colforsin ; pharmacology ; Drugs, Chinese Herbal ; pharmacology ; Heart ; drug effects ; physiopathology ; Heart Failure, Diastolic ; drug therapy ; Isoproterenol ; pharmacology ; Male ; Random Allocation ; Rats ; Rats, Wistar

Objective To compare the effects of vitrification with slow-freezing on the developmental ability of day 3 cleavage stage embryos.Methods Patients who had no less than 4 high quality embryos were included in this study.These embryos were cryopreserved using the methods of vitrification or slow-freezing.In the eryopreserved embryo transfer cycles,the embryos which were cryopreserved using one of the methods were chosen randomly.The developmental ability of embryos was compared between these two groups.Results A total of 80 patients were included in this study with 160 embryos.In the group of slow-freezing,73(91%)embryos were survived and achieved 15(38%)clinical pregnancies.Among these,3 were twins and the implantation rate was 25%(18/73).In the group of vitrification,71(89%)embryos were survived and achieved 19(48%)clinical pregnancies.Among these, 9 were twins and the implantation rate was 39%(28/71),which was significantly higher than the slow- freezing group(P

Objective To investigate the role of metastatic lymph node ratio (MLR) in the evaluation of prognosis of patients with gastric cancer (GC) at different lymph node numbers examined.Methods Clinical data were reviewed retrospectively in a total 535 patients who underwent surgery for GC.Spearman correlation analysis between MLR or number of metastatic lymph nodes (N) and examined lymph node numbers,Kaplan-Meier method was used for comparison survival rates of N stage and MLR stage.A receiver operating characteristic (ROC) curve was used to evaluate the role of N stage and MLR stage in the prognosis of GC patients.Results Metastatic lymph node ratio and number of metastatic lymph nodes correlated with the examined lymph node numbers (r =0.146,r =0.378,P ＜ 0.01,P ＜ 0.001).The 5 year survival rate of MLR0,MLR1,MLR2 and MLR3 patients were 57.5％,69.9％,40.0％ and 21.7％ respectively when examined lymph node numbers ＜ 6 (P ＜ 0.01).The 5-YSR of MLR0,MLR1,MLR2 and MLR3 patients were 86.8％,59.2％,35.8％ and 39.2％ respectively when between 6-10 (P ＜0.001) and the 5-YSR of MLR0,MLR1,MLR2 and MLR3 patients were 88.7％,62.5％,0 and 17.7％ respectively when they ＞ 10(P ＜0.001).AUC of MLR staging was 0.68 ±0.05 when the numbers ＜ 6 (P ＜ 0.001).AUC of MLR staging was 0.72 ± 0.04 at numbers 6-10 (P ＜ 0.001).AUC of MLR staging was 0.79 ± 0.03 when numbers ＞ 10 (P ＜ 0.001).Conclusions MLR was less influenced by lymph node number examined than N.MLR stage has potential superiority to that the N stage in assessing prognosis of GC patients,especially for patients with more than 6 lymph nodes examined.

BACKGROUND: In experimental cerebral ischemia, hypothermia protects the brain, while hyperglycemia aggravates ischemic damage. Clinical studies have also reported worse outcomes in ischemic stroke patients with hyperglycemia or fever and improved outcomes with hypothermic therapy. However, it is not well known what will happen if these opposite effects exist together. METHODS: Sixty male Sprague-Dawley rats were used. Focal cerebral ischemia was induced for 2 hours by an intraluminal thread followed by reperfusion for 4 hours. Rats were divided into 4 groups; 1) normo-glycemic normothermic (NGNT), 2) normoglycemic hypothermic (low temperature) (NGLT), 3) hyperglycemic normothermic (HGNT), and 4) hyperglycemic hypothermic (HGLT) groups (n=15 for each group). Hyperglycemia wasmade by intraperitoneal injection of streptozotocin (60 mg/kg) 3 days before ischemia. The body temperature was maintained at 37+/-1 degrees C in normothermic animals, while lowered at 32+/-1 degrees C in hypothermic animals during the ischemic period. Following reperfusion, 2-mm thick coronal slices were obtained and stained by triphenyltetrazolium chloride. The infarct volume was measured using an image analyzer. RESULTS: Mean glucose levels (mean+/-SD in mg/dl) were 77.9+/-9.3 in NGNT, 77.7+/-11.5 in NGLT, 311.0+/-69.2 in HGNT, and 355.3+/-57.7 in HGLT. Mean infarct volumes (mean+/-SD in mm 3) were 168.2+/-44.2 in NGNT, 66.2+/-24.2 in NGLT, 417.6+/-123.2 in HGNT, and 337.2+/-89.3 in HGLT. The protective effect of hypothermia was less evident in hyperglycemic rats and the aggravating effect of hyper-glycemia was more evident in hypothermic rats. Infarct volume of HGLT was 2-fold bigger than that of NGNT (p<0.05). CONCLUSIONS: Our results suggest that the detrimental effect of hyperglycemia may override the protective effect of hypothermia in transient focal cerebral ischemia.
Animals ; Body Temperature ; Brain ; Brain Ischemia* ; Fever ; Glucose ; Humans ; Hyperglycemia* ; Hypothermia* ; Injections, Intraperitoneal ; Ischemia ; Male ; Rats* ; Rats, Sprague-Dawley ; Reperfusion ; Streptozocin ; Stroke

OBJECTIVETo investigate the pathogenesis of globozoospermia, fertilization ability of round-headed sperm, and the application value of assisted oocyte activation in intracytoplasmic sperm injection (ICSI) for the wives of glohozoospermia men.

METHODSWe collected oocytes from the wives of 2 globozoospermia patients and randomly divided them into two groups after ICSI to receive calcium ionophore A23187-activation and conventional treatment, respectively. We reviewed the relevant literature published at home and abroad, and discussed the etiology of globozoospermia, fertilization ability of round-headed sperm, and treatment options for this disease.

RESULTSQuality embryos were obtained in the A23187-activation group while no fertilized oocytes, oocyte cleavage, quality embryos, or blastular formation were found in the conventional treatment group. Both women achieved pregnancy and gave birth to healthy neonates after transfer of the quality embryos from the A23187-activation group.

CONCLUSIONCalcium ionophore A23187 can be applied to ICSI for the wives of globozoospermia men and bring about desirable clinical outcomes. Meanwhile, attention should be paid to its safety.

Calcimycin ; therapeutic use ; Calcium Ionophores ; therapeutic use ; Female ; Humans ; Infertility, Male ; drug therapy ; Male ; Oocytes ; Pregnancy ; Sperm Injections, Intracytoplasmic ; Spermatozoa ; abnormalities

Objective To explore the therapeutic efficacy of levofloxacin combined with anti-tuberculosis drugs and thoracic catheterization for the treatment of tuberculous pleuritis.Methods Patients who were admitted to Departments of Infectious Diseases of Hanzhong Central Hospital and Ankang Central Hospital between February 2014 and August 2016 for initial treatment of tuberculous pleuritis were included in the study,they were divided into groups A,B,C and D.Group A received 2HRZE + 7HR regimen combined with conventional drainage;group B received 2HRZE+ 7HR regimen combined with thoracic catheterization;group C received 2HRZEV + 7HR regimen combined with thoracic catheterization;group D received 2HRZEV + 10HR regimen combined with thoracic catheterization.groups B,C and D received thoracic catheterization,normal saline 20mL and urokinase 100,000U were given through the drainage tube.Results A total of 172 patients with newly diagnosed tuberctlous pleurisy were received for treatment.There were 45,53,38,and 36 cases in group A,B,C,and D respectively.The total effective rate of therapy for pleural effusion in group A was lower than that in group B(64.44％ vs 90.57％,x2 =9.863,P＜ 0.05);after two month therapy,total effective rate of therapy for pleural effusion in group B was lower than that in group C (18.87％ vs 39.47％,x2 =4.716,P＜0.05);at the end of therapy,total effective rate in group C was lower than that in group D (60.53 ％ vs 83.33 ％,x2 =4.731,P＜0.05).Conclusion For initial treatment of patients with tuberculous pleuritis,2HRZEV + 10HR antituberculosis regimen combined with thoracic catheterization and urokinase infusion can significantly improve the clinical symptoms and recovery rate of tuberculous pleuritis,facilitate drainage of pleural effusion and prevent pleural thickening,adhesion and encapsulation.

Diagnosis, Differential ; Electroencephalography ; Female ; Follow-Up Studies ; Humans ; Infant, Newborn ; Male ; Parasomnias ; diagnosis ; pathology ; Remission, Spontaneous ; Seizures ; diagnosis ; pathology ; Sleep Stages ; physiology

OBJECTIVETo compare the effects of the cryoprotectant containing glucose and that containing sucrose on the motility of post-thaw human sperm.

METHODSThe cryoprotectant containing glucose and that containing sucrose were applied to 50 semen samples and the motility of the post-thaw human sperm was compared before and after cryopreservation and between the study groups.

RESULTSThe forward motility and total motility of the sperm were (58.4 +/- 5.7)% and (63.4 +/- 6.1)% before cryopreservation, (43.8 +/- 7.6)% and (48.4 +/- 7.6)% after thawing with the cryoprotectant containing glucose, and(42.6 +/- 8.9)% and (48.0 +/- 8.5)% after thawing with the cryoprotectant containing sucrose. Decreased sperm motility was observed after cryopreservation, with statistic significance (P < 0.01). There was no significant difference in the forward and total motility of the post-thaw sperm between the two cryoprotectants.

CONCLUSIONCryopreservation inflicts obvious damage on sperm. Sucrose is a feasible sperm cryoprotectant.

Adult ; Cryopreservation ; methods ; Cryoprotective Agents ; pharmacology ; Glucose ; pharmacology ; Humans ; Male ; Semen Preservation ; methods ; Sperm Motility ; drug effects ; Spermatozoa ; drug effects ; physiology ; Sucrose ; pharmacology

CD40/CD40L interactions play a pivotal role in T cell activation, and take part in many physiologic and pathologic procedures and different levels. In this article, stable CHO transformants secreting human CD40-Ig fusion protein were established through transfection and selection with Lipofectamaine and G418, respectively. In order to obtain great valume of recombinant protein, big batch serum-free cultures of engineered CHO cells were performed in roller-bottle using CHO-II-SFM medium. After cultures, the cell-culture supernatants were harvested, concentrated through ultra-filtration, and finally purified by affinity choromatography with Protein G Sepharose Fast Flow. Human peripheral bloods were collected freshly and seperated with Ficoll, CFU-T was cultured in semi-solid culture system with peripheral blood mononuclear cells (PBMNC). Effect of human CD40-Ig fusion protein on the formation of CFU-T was observed in vitro. The results showed that the yield of human CD40-Ig fusion protein was 30 mg in total 3 liter CHO-II-SFM culture supernatant, and it supposed that the expression level of CD40-Ig in CHO cells was more than 10 micro g/ml. The purity of purified fusion protein is above 95%. Furthermore, compared with human IgG, human CD40-Ig fusion protein significantly inhibited the formation of CFU-T at dose 0.25, 1.0, 4.0, and 10 micro g/ml, it lays a good foundation to evaluate its potential functions in vivo.