Fifty oocytes were studied with light microscope and electron microscope. The shape of oocyte is round, about 50—100?m in diameter. The nucleus is large and spheroidal in shape. The diameter of the nucleus is 15—20?m. All the oocytes are surrounded by zona pellucida and follicular cells. Call-Exner bodies are seen between these follicular cells. A great number of microvilli are present on the surface of oocytes. The paired membranes of nucleus envelope is clearly seen. There are a lot of nuclear pores on nuclear envelope, about 4,000/mm~2. The nuclear pore complex is disinct. The cytoplasm of oocyte abounds with organelles which are distributed regularly. The cristae of mitochondria are short and scarety. The mitochondrial matrix becomes very dense. The cavities of endoplasmic reticulum are distended. Well developed Golgi complex are usually found at the periphery of oocytes. Annulate lamellae consist of parallel flat sacs with a length of 1.5—2.0?m. There are structures like nuclear pores on the flat sac. Some bunchs of fibrous material in the cytoplasm of oocytes are called yolk plates Each yolk plate consists of a regular alternating dark and light bands with a periodicity of 20nm. Cortical granules are round with a diameter of 0.2—0.4?m. They are wrapped up in a membrane. Their electron density is high.

ObjectiveTo evaluate the effect of N(2)-L-alanyl-L-glutamine (Ala-Glu) on perioperative insulin resistance in patients undergoing radical colon cancer operation.MethodsSixty ASA Ⅰ or Ⅱ patients of both sexes aged 35-75 yr with BMI of 18.5-25.0 kg/m2 undergoing redical colon cancer operation under general anesthesia were randomly divided into 3 groups ( n =20 each):control group(group C) ; vehicle group (group Ⅴ) and group Ala-Glu.Ala-Glu 22.5 ml/kg was administered iv at 24 h before and 1 h after operation in group AlaGlu,while in groups C and V equal volume of normal saline and vehicle were given iv instead of Ala-Glu.Venous blood samples were taken at 24 h before operation (T1),30 main before ( T2 ) and 3 h after induction of anesthesia (T3) and 1 and 24 h after operation (T4,T5 ) for determination of blood concentrations of glucose (BG),insulin ( INS)-,TNF-α and free fatty acid (FFA).Insulin resistance ( HOMA-IR =BG × INS ÷ 22.5) and insulin sensitivity index (ISI =1 ÷ (lgBG + lgINS) ) were calculated.The time when the patients passed flatus,the days of hospitali-zation after operation,and the incidence of insulin resistance were recorded.ResuitsAla-Glu significantly decreased blood concentrations of BG,INS,TNF-α,FFA and HOMA-IR and increased ISI in group Ala-Glu as compared with groups C and V.The patients passed flatus earlier after operation and postoperative hospital stay was shorter and the incidence of insulin resistance was lower in group Ala-Glu than in groups C and V.There was no significant difference in all the indexes between group C and group V.ConclusionN (2)-L-alanyl-L-glutamine can attenuate perioperative insulin resistance in patients undergoing colon cancer resection and is helpful to patient' s recovery,and the decrease in the concentrations of TNF-α and FFA may be involved in the mechanism.

6 cases of gamete intra-fallopian transfer (GIFT) were performed in cooperation with the Department of Gynaecology and Obstetrics of The Third Teaching Hospital of our university in 1987. All the six women aged less than 41 years and have failed to be pregnant as long as ten years. One fallopian tube is patent at least. Two of them were pregnant after operation of GIFT. The rate of success was 33％. We used Ham's F10 medium mixed with different concentrations of fetal cord serum or mother's serum to prepare respectively follicular flushing medium (FM), sperm washing medium (SW), growth medium (GM) and transfer medium (TM). Clomiphene citrate and human menopausal gonadotropin (HMG) were used for inducing growth of follicle. HCG was given by intramuscular injection when two or more follicles were measured 16 mm in diameter by pelvic ultrasound. 32-34 hours later, follicular aspiration were performed by abdominal operation and the oocytecorona-cumulus complexes (OCCCs) were collected and trimmed under stereo microscope. The semen samples were obtained by means of masturbation 2-3 hours before operation. After centrifuged twice, the liquified semen was mixed with SW and incubated with 5％ CO_2 in air at 37℃ until GIFT.The OCCCs and 25?l processed semen were loaded into a special catheter and transferred into one fallopian tube. Then the transfer procedure was repeated for the other fallopian tube. The level of HCG in blood was examined from day 11 after transfer in order to diagnose pregnancy.

Objective To detect the mutations in ATP2C1 gene in 3 Chinese Hailey-Hailey-disease (HHD) families and 1 sporadic HHD patient.Methods Three Chinese HHD families and 1 sporadic HHD patient were recruited into this study with informed consent.Blood samples were taken from the patients with HHD,unaffected individuals in the HHD families and 100 unrelated normal human controls.Genomic DNA was extracted from these blood samples.All the exons and exon-intron boundaries of the ATP2C1 gene were amplified by PCR followed by direct sequencing via dye-termination chemistry.Results Three novel missense mutations in ATP2C1 gene were identified,including a 2048 G→A mutation in exon 20 causing the substitution of arginine by lysine at position 619 in the patients from HHD family 1,853A→C mutation in exon 8 causing the substitution of threonine by proline at position 221 in the patients from family 2,and 2323T→C mutation in exon 23 causing the substitution of tyrosine by histidine at position 711.None of these mutations were found in patients from the HHD family 3,unaffected individuals from the HHD family 1 and 2,or the unrelated normal human controls.Conclusion Three novel missense mutations are identified in the ATP2C 1 gene of patients with HHD.

Objective To investigate the role of CCL2 in pain facilitation and spinal mechanisms in the rat model of bone cancer pain.Methods The bone cancer pain model was developed by inoculating.Walker 256 mammary gland carcinoma cells into the rat tibia medullary cavity.SD female rats were divided into 5 groups randomly ( n =8):sham group( group Ⅰ),sham + CCL2 antibody group( group Ⅱ),BCP group( group Ⅲ),BCP +control lgG group ( group Ⅳ),BCP + CCL2 antibody group ( group Ⅴ ).VonFrey threshold was measured one day before operation and 1 st,3 rd,5th,7th,10th,14th,21 st after operation.CCL2 antibody or control lgG was injected intrathecally from 10th to 12th day.The expression of the spinal Iba-1 ( microglial marker) in rat lumbar4-5 was detected by immunohistochemistry assay.Results From the 10th to 21st day after operation,the PMWT of group Ⅲ rats were ( 1.78 ±0.38)g,( 1.70 ±0.17)g,( 1.35 ±0.07 )g;group Ⅳ rats were (2.99 ±0.67)g,(2.52 ±0.75)g,(1.13±0.07)g ; and group Ⅴ rats were (5.88±0.66)g,(7.81 ±0.75)g,(6.19±0.53)g.Compared with group Ⅲ,the PMWT of group Ⅴ was remarkly higher (P＜0.01) ; group Ⅳ had no obvious statistical significance (P＞0.05).At the 14th day after operation,the MOD of group Ⅲ,Ⅳ and Ⅴ rats were (151.3 ±10.8 ),( 149.2 ± 10.6),(74.5 ± 5.0),Compared with group Ⅲ,the MOD of group Ⅴ was significantly increased (P＜0.01 ),group Ⅳ had no obvious statistical significance (P ＞ 0.05 ).Conclusion Intrathecal injection of CCL2 antibody can remarkly attenuate established pain facilitation of tibial bone cancer pain rats,and significantly suppress the expression of Iba-1.It suggests that CCL2 is involved in the bone cancer pain via activation of spinal microglia.

Objective To analyze the changes in peripheral blood monocyte subpopulations in patients with primary, secondary and latent syphilis. Methods Flow cytometry was used to detect CD14highCD16- and CD14+CD16+ monocyte subpopulations in peripheral blood from 58 patients with untreated syphilis, including 36 cases of latent syphilis,8 cases of primary syphilis and 14 cases of secondary syphilis, as well as from 65 normal human controls. Restflts Compared with the normal controls, the proportion of CD14+CD16+ monocytes among total monocytes was significantly elevated (12.0% ± 5.0% vs 6.0% ± 3.3%, t = 7.25, P < 0.01), while that of CD14highCD16- monocytes was down-regulated (88.0% ± 5.1% vs 94.0% ± 3.5%, t = -7.20, P < 0.01). No statistical difference was observed in the proportion of CD14+CD16+ or CD14hhighCD16- monocytes among the patients with primary syphilis, secondary syphilis and those with latent syphilis (all P > 0.05). Conclusions The changes in peripheral blood monocyte subpopulation in patients with untreated syphilis may be associated with the permanent infection of Treponema pallidum, but have no obvious correlation with clinical stage of syphilis.

Summary: In order to investigate the effects of aconitine on [Ca2+] oscillation patterns in cultured myocytes of neonatal rats, fluorescent Ca2+ indicator Fluo-4 NW and laser scanning confocal microscope (LSCM) were used to detect the real-time changes of [Ca2+] oscillation patterns in the cultured myocytes before and after aconitine (1.0 μmol/L) incubation or antiarrhythmic peptide (AAP) and aconitine co-incubation. The results showed under control conditions, [Ca2+] oscillations were irregular but relatively stable, occasionally accompanied by small calcium sparks. After incubation of the cultures with aconitine, high frequency [Ca2+] oscillations emerged in both nuclear and cytoplasmic regions, whereas typical calcium sparks disappeared and the average [Ca2+] in the cytoplasm of the cardiomyocyte did not change significantly. In AAP-treated cultures, intraecllular [Ca2+] oscillation also changed, with periodic frequency, increased amplitudes and prolonged duration of calcium sparks. These patterns were not altered significantly by subsequent aconitine incubation. The basal value of [Ca2+] in nuclear region was higher than that in the cytoplasmic region, in the presence or absence of drugs, the [Ca2+] oscillated synchronously in both the nuclear and cytoplasmic regions of the same cardiomyocyte. It was concluded that although oscillating strenuously at high frequency, the average [Ca2+] in the cytoplasm of cardiomyocyte did not change significantly after aconitine incubation, compared to the controls. The observations indicate that aconitine induces the changes in [Ca2+] oscillation frequency other than the Ca2+ overload.

Objective To explore the effect of polyethylene glycol-polyethyleneimine (PEG-PEI) serving as a non-viral vector in delivering small interfering RNA (siRNA) into C17.2 neural stem cells (NSCs) in vitro. Methods Complexes of PEG-PEI and siRNA targeting Nogo receptor were prepared, and their characterizations were estimated by measurements of particle size and zeta potential,and the complex abilities of PEG-PEI/siRNA complexes were observed by gel retardation assay. In addition, with liposome complex system (Lipofectamine 2000/siRNA) as positive control, the transfection efficiency of PEI-PEG/siRNA complexes at different N/P ratios (cationic nitrogen/siRNA phosphate molar ratio) was detected by flow cytometry. Results The siRNA molecules were condensed by PEG-PEI to form nanoseale complexes. As the proportion of N/P ratio enhancing, the surface potential of nanoparticles gradually increased and the particle sizes of PEI-PEG/siRNA complexes showed a decreasing trend. Gel retardation electrophoresis suggested that siRNA could be fully composited with PEG-PEI as a result of the coulombic foree between them. Meanwhile, flow cytometry experiments revealed that the transfection efficiency of PEG-PEI mainly depended on N/P ratios of the nanoparticles,and the highest one was obtained at N/P=15 ([78.72±8.18)]％). Conclusion PEG-PEI might be a prospective candidate for siRNA delivery system, which enjoys its value in NSC gene therapy.

Objective To compare the differences on therapeutic approach and short-term outcomes between male and female patients with ST-elevation myocardial infarction ( STEMI ).Methods Data of Chinese STEMI patients were retrospectively analyzed from a global multicenter clinical trial of reviparin and metabolic modulation in acute myocardial infarction treatment evaluation (CREATE). The patients were divided into two groups according to gender and difference on demographic,baseline clinical characteristics at admission, reperfusion and drug therapy and 30-day all-cause mortality, re-infarction, stroke,hemorrhage,heart failure and combined end points were compared.Results Of the 7431 patients,29.1％were female.Female patients were older than male patients [ ( 68.2 ± 9.1 ) years vs.( 60.3 ± 12.1 )years ].On admission,heart rate,diastolic blood pressure and Killip class were higher and the delay between onset of chest pain and arrival at hospital was longer in female patients than in male patients ( all P ＜0.01 ).Male patients often presented MI in anterior leads while female patients often presented MI in inferior and lateral leads.History of diabetes mellitus,hypertension and heart failure was significantly higher in female than in male patients ( all P ＜0.01 ).Incidence of high blood glucose was higher in female while high blood potassium was higher in male patients ( P ＜ 0.01 ).Rate of reperfusion therapy was lower and the use aspirin and diuretic was more frequent in female patients than in male patients,while frequency of clopidogrel,Glycoprotein Ⅱ b/Ⅲ a receptor inhibitor,β-blockers,angiotensin converting enzyme inhibitor,lipid-lowering drug use was significantly higher in male than in female patients ( all P ＜ 0.01 ). Multivariate logistic regression analysis showed that female gender was a predictor for less PCI therapy (P ＜ 0.01 ).Thirty-day all cause mortality( OR =1.425,95％ CI:1.163 - 1.747,P ＜ 0.01 ) and combined end points ( OR =1.193,95％CI:1.010- 1.410,P =0.04) were significantly higher in female patients than in male patients.Conclusions There are gender-related differences on therapeutic approach and short-term outcome in Chinese STEMI patients.The unfavorable demographic and baseline clinical profile could partially explain the less reperfusion therapy rate and worse prognosis in female patients.