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 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.

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

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

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

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 study the mechanism of the effect of low-frequency rotary constant magnetic field on high-fat and high-protein diet-induced fatty liver in rats.

METHODSFatty liver model was established in SD rats by feeding on a high-fat and high-protein diet daily. The enzyme activity changes in the serum and liver homogenate were detected at 10, 14, and 18 weeks, and the pathological changes of the liver were observed with optical and electron microscopy.

RESULTSIn magnetic field intervention group, the concentration of alanine aminotransferase and aspartate transaminase were significantly decreased, and the activity of lipoprotein lipase, hepatic lipase, superoxide dismutase and the concentration of malondialdehyde in the liver homogenate were significantly increased. Under optical microscope and electron microscope, the rats in the model group showed diffusive adipose degeneration in the hepatic cells with large lipid droplets, which became large vacuoles after fat extraction, indicating fatty necrosis. In magnetic field intervention group, remarkably smaller lipid droplets and lessened hepatic cell adipose degeneration were observed.

CONCLUSIONLow-frequency rotary constant magnetic field has beneficial effect on fat metabolism, leading to reduced lipid peroxidation and structural recovery of the degenerated hepatic cells.

Animals ; Dietary Fats ; administration & dosage ; Dietary Proteins ; administration & dosage ; Fatty Liver ; etiology ; pathology ; therapy ; Magnetic Field Therapy ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley

Embryonic hematopoiesis in mammals is characterized by successive temporal and spatial changes. Previous investigations indicate that in vitro differentiation of embryonic stem cells (ES cells) derived from 129 mice can mimic embryonic hematopoiesis to some extent. To investigate the in vitro hematopoietic differentiation capacity of ES cells derived from C57BL/6 mice, the authors initially established the murine ES cell line with standard identification methods employed. Next, two-step culture system was utilized for embryoid bodies formation and the appearance of different hematopoietic precursors was confirmed by CFC assay, cellular chemical staining as well as RT-PCR. The results demonstrated that the ES cell line MES-1 fulfilled the criteria of ES cell line and its progeny after in vitro differentiation included primitive and definitive erythrocyte precursors, mixed colony-forming cells and granulocyte/macrophage colony-forming cells. RT-PCR analysis revealed the molecular consistence of transcription factors and hematopoietic markers with cellular event. In conclusion, MES-1 established from C57BL/6 mice was able to differentiate in vitro to a variety of hematopoietic precursors, thus could partly recapitulate embryonic hematopoiesis.
Animals ; Cell Culture Techniques ; methods ; Cell Differentiation ; genetics ; Cell Line ; Colony-Forming Units Assay ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; cytology ; Erythroblasts ; cytology ; metabolism ; Erythroid Precursor Cells ; cytology ; metabolism ; Erythroid-Specific DNA-Binding Factors ; Gene Expression ; Hematopoietic Stem Cells ; cytology ; metabolism ; Mice ; Mice, Inbred C57BL ; Mice, Inbred Strains ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells ; cytology ; metabolism ; Time Factors ; Transcription Factors ; genetics ; Vascular Endothelial Growth Factor Receptor-2 ; genetics