Objective To develop a method for treating infertile women with polycystic ovary syndrome(PCOS)by ooeyte in vitro maturation(IVM).Methods On day 3～4 of menstrual cycle,infertile women with PCOS were administrated with(100～150IU)FSH/day for 3 days.On day 8～10,when the follicle diameter reached 10～14mm and endometrium＞5mm,the immature oocytes weFe picked up and cultured in vitro for 28～31 hours.Then,ICSI and embryo transfer were performed.Results 3 clinic pregnancies were established among 12 infertile women.the pregnancy rate was 25.0%.Conclusion This research indicates that IVM is a feasible technology for the infertile women with PCOS.

Objective To evaluate the role of Toll-like receptor 2/nuclear factor-κB(TLR2/NF-κB)signaling pathway pretreatment in ventilator-induced lung injury(VILI). Methods Thirty male Sprague-Dawley(SD)rats were randomly divided into three groups by using random number scale,with 10 rats in each group. Group A:rats were given 200μL of TLR2 monoclonal antibodies(TLR2mAb,10μg/kg)by slow instillation through tracheal catheter, and then ventilated with a high tidal volume(VT)of 40 mL/kg. Group B:ventilated with a normal VT of 8 mL/kg. Group C:rats were tracheally instilled with 10 μg/kg of TLR2mAb devoid of biologic activity,and then ventilated with a high VT of 40 mL/kg. The rats were mechanically ventilated for 4 hours,the lung wet to dry weight ratio(W/D)was calculated. The changes in pathology and ultrastructure in lung tissue were observed with microscope. Enzyme linked immunosorbent assay(ELISA)was performed to determine the concentration of interleukins(IL-1β,IL-6)and tumor necrosis factor-α(TNF-α)in serum and brconchoalveolar lavage fluid(BALF). Real-time fluorescent quantitation reverse transcription-polymerase chain reaction(RT-PCR)was used to assess the mRNA expressions of TLR2, NF-κB and myeloid differentiation factor 88(MyD88)in lung tissue. Results No obvious pathological changes in lungs were found in group A and group B,and no obvious damages to ultra-microstructure were found in lung macrophages, typeⅠepithelial cell and typeⅡepithelial cell. In group C,pathological changes were observed,including pulmonary alveoli fusion,alveoli septum thickening,inflammatory cells infiltration,and damages to ultrastructure of lung macrophage,damage to cell membrane of typeⅠepithelial cells and typeⅡepithelial cells,vacuoles in cytoplasm, damage to organelle,and even pyknosis and perinuclear cistern thickening. Compared with group C,W/D ratio and mean concentration of inflammatory cytokines in serum and BALF showed a significant decrease in group A and B〔W/D ratio:1.151±0.026,1.128±0.048 vs. 1.403±0.062;concentration of IL-1βin serum(ng/L):37.05±5.61, 34.52±4.31 vs. 51.45±8.18;concentration of IL-6 in serum(ng/L):53.65±5.16,55.77±5.62 vs. 89.96±7.08;concentration of TNF-αin serum(ng/L):71.93±13.29,67.36±11.42 vs. 96.20±11.60;concentration of IL-1βin BALF(ng/L):56.48±6.16,54.44±7.26 vs. 99.77±8.41;concentration of IL-6 in BALF(ng/L):172.44±21.26, 163.47±18.70 vs. 216.22±23.90;concentration of TNF-α in BALF(ng/L):235.81±42.75,231.72±40.38 vs. 374.85±69.61,all P<0.01〕,but there were no significant differences between group A and group B(all P>0.05). The mRNA expressions of TLR2,MyD88,and NF-κB were significantly decreased in group A and group B compared with those in group C〔TLR2 mRNA(2-ΔΔCt):1.021±0.287,0.938±0.196 vs. 3.862±0.871;MyD88 mRNA (2-ΔΔCt):1.235±0.277,1.300±0.306 vs. 3.618±1.107;NF-κB mRNA(2-ΔΔCt):0.519±0.036,1.043±0.170 vs. 20.280±9.466,P<0.05 or P<0.01〕,but there was no significant difference among the parameters mentioned above between group A and B(all P>0.05). Conclusion To some extent,pre-intervention with TLR2mAb to block the TLR2/NF-κB signal pathway can inhibit the release of pro-inflammatory factors,and regulate the VILI.

Objective To observe the influence of patient′s age, and the number of transferred-good-quality-embryos on multiple gestation rates in in vitro fertilization and embryo transfer(IVF-ET) cycles. Methods In this retrospective study, a total of 4395 patients who transferred fresh embryo between Jan 2004 and Nov 2006 was analyzed. According to the age, the patients were divided into 2 groups: aged < 35 (3442 cycles) or aged ≥135(953 cycles). We regularly transferred 2 -3 embryos. If the patients had only one embryo, one was transferred. And those patients who had only 2 embryos, even if they were more than 35 years old or it would be the second time for them to transfer, were transferred 2 embryos. The influence of female age and the number of good quality embryos transferred on the multiple gestation rates in IVF-ET cycle was analyzed. Results (1)The multiple gestation rate of the groups of 1 good quality embryo,2 good quality embryos, or 3 good quality embryos transferred were 21.08% (35/166), 31.41% (413/1315), and 42. 37% (75/177), respectively in women aged < 35, with a significant difference between them. The pregnancy rates of these groups were 29. 64% (166/560) ,51.63% (1315/2547) ,and 52. 84% (177/335), respectively; there were no significant differences between 2 good quality embryos transferred group and 3 good quality embryos transferred group. (2) The multiple gestation rates of the groups of 1 good quality embryo,2 good quality embryos, or 3 good quality embryos transferred were 19. 51% (8/41) ,20. 65% (19/ 92) ,and 40.66% (74/182), respectively, in women aged ≥ 35; there were no significant differences between 1 good quality embryo transferred group and 2 good quality embryos transferred group. The pregnancy rates of these groups were 19. 07% (41/215), 33.70% (92/273), and 39. 14% (182/465), respectively; there were no significant differences between 2 good quality embryos transferred group and 3 good quality embryos transferred group. (3) The pregnancy rate of the patients aged <35 [48. 17% ( 1658/ 3442) ]was significantly higher than in women aged ≥35[33.05% (315/953) ]. Conclusion The transfer of 2 good quality embryos results in similar pregnancy rates and significantly reduced multiple gestation rates when compared to the transfer of 3 good quality embryos in women regardless of their ages.

Objective To investigate the changes in the expression of macrophage migration inhibitory factor (MIF) mRNA in a rat model of ventilator-induced lung injury.Methods Thirty adult male Sprague-Dawley rats,weighing 2β5-260 g,were randomly divided into 3 groups (n =10 each) using a random number table:control group (group C),small tidal volume (VT) mechanical ventilation group (group S) and large tidal volume mechanical ventilation group (group L).The animals were anesthetized with intraperitoneal ketamine 100 mg/kg,midazolam 0.2 mg/kg and atropine 1.0 mg/kg.The rats were tracheostomized and spontaneous breathing was maintained in group C,while the rats were tracheostomized and mechanically ventilated for 4 h in groups S and L.The tidal volume was 7 ml/kg (group S) or 40 ml/kg (group L),I ∶ E was 1 ∶ 1,RR was 80 bpm and FiO2 was 100％.At 4 h of spontaneous breathing or mechanical ventilation,broncho-alveolar lung lavage fluid (BALF) was collected for determination of the total protein concentration,white blood cell (WBC) counts and concentrations of MIF,IL-6 and IL-1β (by ELISA).Then the rats were sacrificed and the lungs removed for microscopic examination and for determination of wet to dry lung weight ratio (W/D ratio) and expression of MIF mRNA (by RT-PCR).Results Compared with C and S groups,WBC counts,concentrations of total protein,MIF,IL-6 and IL-1β in BALF,and W/D ratio and expression of MIF mRNA in lung tissues were significantly increased in group L (P ＜ 0.05).There was no significant difference in the indexes mentioned above between group C and group S (P ＞ 0.05).The pathological changes occurred in group L.Conclusion The up-regulation of MIF mRNA expression in lung tissues may be involved in the development of ventilator-induced lung injury in rats.

Objective To evaluate the efficacy and safety of levetiracetam (LEV) monotherapy on children with epilepsy.Methods Forty-one children (26 cases were male,15 cases were female) with epilepsy aged 7 months to 13 years were treated with LEV as monotherapy.These patients were selected from Department of Neurology ,Wuhan Children′s Hospital, from Aug.2007 to Aug.2009.The starting do-sage of LEV was (13.6?4.7) mg?kg-1?d-1,twice daily,and its objective dosage was (25.7?7.5) mg?kg-1?d-1,twice daily.LEV monotherapy was investigated by a self-controlled and open-label research,and the follow-up period ranged from 6 months to 2 years.Results The effective rate was 68.3% (28 cases),with 39.0% (16 cases) achieving seizure freedom in LEV monotherapy of children with epilepsy.Thirteen patients (31.7%) had poor efficacy in reduction of seizures,7 patients (17.1%) discontinued LEV monotherapy due to either an inadequate seizure control or aggravated seizures.Fifteen patients (36.6%) had the therapy-related adverse events in LEV monotherapy,including gastrointestinal dysfunction (5 cases),irritability (5 cases),dizziness (2 cases) and somnolence (2 cases).The adverse effects appeared in 2-4 weeks of early LEV therapy and were spontaneously disappeared in 1 week to 1 month of continuing therapy.Conclusions The LEV monotherapy is effective and safe for the control of partial and generalized seizures in children with epilepsy.LEV appears to be a broad-spectrum,first-line anti-epileptic drug in treatment of children with epilepsy.

Objective To investigate the clinical characteristics and diagnostic main points of poliomyelitis-like syndrome(PS).Met-hods The clinical data of 15 children with PS were analyzed retrospectively including nervous system manifestation,results of lab andauxiliary examination,diagnosis and reason of misdiagnosis,treatment and progress.Results PS in all 15 children was characterized by mean age of onset(3.3 years),and forerunner infection(in 11 cases,73%)with acute upper respiratory infection of diarrhea 1-2 weeks pre-onset or during onset.All cases were acute flaccid paralysis,12 cases(80%) of which were only one limb.The positive outcome of serologic examination in PS consisted of 4 cases in coxsackie virus-IgM,1 case in EB virus-IgM,1 case in herpes simplex virus-IgM and 1 case in mycoplasma-IgM.All children showed the electromyologram changes in nerve damage.The muscle force of 4 cases increased one grade.Conclusions Children with PS are characterized by the age of onset under 5 years old,acute flaccid paralysis(mostly affected one limb),and the most pathogen being enterovirus.The electromyologram examination can help establish a definite diagnosis in PS.

Objective To investigate the role of Toll-like receptor9 (TLR9)-myeloid differentiation factor 88 (MyD88) signal pathway in alveolar macrophages in ventilator-induced lung injury (VILI).Methods 30 adult male Sprague-Dawley (SD) rats were randomly assigned to three groups (with 10 rats in each group).Group A was the control group,with spontaneous respiration after tracheostomy.Rats in group B received mechanical ventilation for 4 hours with normal tidal volume (VT) 7 ml/kg after tracheostomy,and group C rats received mechanical ventilation with VT 40 ml/kg for 4 hours.After termination of ventilation,examination with transmission electron microscopy was performed to observe the ultrastructure changes in alveolar epithelial cell type Ⅱ (AEC Ⅱ) of the lung.Lung wet/dry ratios (W/D) and total protein concentration,the concentration of interleukins (IL-6 and IL-1 β) in bronchoalveolar lavage fluid (BALF) were determined.The protein and mRNA expressions of TLR9,MyD88 and nuclear factor-κB (NF-κB) in alveolar macrophages were assayed by Western Blot and real-time reverse transcription-polymerase chain reaction (RT-PCR).Results The ultrastructure of AEC Ⅱ in the group A and group B was almost normal,whereas the chromatin of the nuclei,the lamellar corpuscles in the cytoplasm,the cell membrane and the microvilli of the AEC Ⅱ in the group C showed injurious changes in various degrees.When the group C was compared with the group A and the group B,it was shown that the W/D ratios (5.54 ± 0.17 vs.4.58 ± 0.17,4.69 ± 0.16) and total protein concentration (g/L:6.33 ± 0.61 vs.0.45 ± 0.05,0.47 ± 0.04),IL-6 (μg/L:1.989 ± 0.103 vs.1.033 ± 0.061,1.010 ± 0.069) and IL-lβ (ng/L:2.79 ±0.25 vs.1.05 ±0.15,1.23 ±0.22) in BALF,the protein expressions of TLR9,MyD88 and NF-κB [TLR9 (A value):0.770 ±0.042 vs.0.300 ±0.027,0.310 ±0.037; MyD88 (A value):0.950 ±0.091 vs.0.560 ±0.082,0.580±0.084; NF-κB(A value):1.020 ±0.076 vs.0.740 ±0.052,0.700 ±0.076] in alveolar macrophages were all increased significantly,and all of which showed significant difference (P＜0.05 or P＜0.01).The mRNA levels of TLR9,MyD88 and NF-κB in alveolar macrophages in the group B were (1.13 ± 0.32),(1.18 ± 0.33),and (1.11 ± 0.22) folds of those of the group A,respectively,but there were no significant differences (all P＞0.05).While the mRNA levels of TLR9,MyD88 and NF-κB of alveolar macrophages in the group C were (8.66 ± 0.69),(6.41 ± 0.53) and (5.29 ± 0.71) folds of those of the group A,respectively,and all of them showed significant difference (all P＜0.01).Conclusion TLR9-MyD88 signaling in alveolar macrophages plays a role in pathogenesis of VILI.

OBJECTIVE To explore the role of P38MAPK inhibitor on the soft palate reconstruction of the rats with chronic intermittent hypoxia. METHODS The animals were divided into normal control group, hypoxia control group and SB203580+hypoxia group (every group 20 rats). After 5 weeks, the expressions of p38MAPK and p-p38MAPK protein on the soft palate of the rats were detected with immunohistochemical techniques and western blot. RESULTS Compared with the normal control group, the soft palate tissue thickness of the hypoxia group were increased most obviously; Levels of p38MAPK were increased in hypoxia control group; Compared with the hypoxia control group, the levels of p-p38MAPK group were decreased in SB203580+hypoxia group. CONCLUSION p38MAPK may play important roles in the soft palate reconstruction of the rats with chronic intermittent hypoxia.

Objective To investigate the role of Ras-related C3 botulinum toxin substrate 1/mitogen-activated protein kinase/extracellular signal-regulated kinase (Rac 1/MAPK/ERK) signal pathway in rats with ventilator induced lung injury (VILI) and its mechanism.Methods Thirty Sprague-Dawley (SD) rats were randomly divided into spontaneous respiration group,normal tidal volume (VT) group and high VT group with 10 rats in each group.The rats in spontaneous respiration group were kept their spontaneous breathing.The rats in normal VT group and high VT group were performed tracheal intubation after tracheostomy,and underwent mechanical ventilation on bilateral lungs with 6 mL/kg and 40 mL/kg VT respectively with maintenance anesthesia.After 4-hour ventilation,heart blood,bronchoalveolar lavage fluid (BALF) and lung tissues were harvested.The levels of interleukins (IL-1β,IL-6),tumor necrosis factor-α (TNF-α),myeloperoxidase (MPO) and macrophage inflammatory protein-2 (MIP-2) in serum and BALF were determined by enzyme linked immunosorbent assay (ELISA).Lung wet/dry radio (W/D) was determined.The lung tissues were stained with hematoxylin and eosin (HE),and pathological changes were observed,and pathological scores were evaluated.The ultra structure changes in type Ⅱ alveolar epithelial cells (AEC Ⅱ)were observed with transmission electron microscope.The positive expressions of phosphorylation of extracellular signal-regulated kinase (p-ERK) were determined by immunohistochemistry,and those of Racl and F-actin were determined by immunofluorescence.The mRNA expressions of ERK and Rac1 were determined by real-time fluorescent quantitation reverse transcription-polymerase chain reaction (RT-qPCR),and protein expressions of Rac-1,p-ERK and F-actin were determined by Western Blot.Results ① Compared with spontaneous breathing group,lung W/D in both mechanical ventilation groups was significantly increased,with more significant increase in the high VT group (6.64 ± 0.88 vs.1.79 ± 0.36,P ＜ 0.01).② There was no obvious pathological changes in the lung tissue and AEC Ⅱ of the spontaneously breathing group.In the normal VT group,there was slight edema and infiltration of inflammatory cells;AEC Ⅱ had less lamellar bodies and uniform distribution of the villi of the alveolar epithelium.In the high VT group,the edema of the lung tissue,the widening of the pulmonary septum,the alveolus congestion,the infiltration of inflammatory cells,and alveolar structure disorder were found;and AEC Ⅱ was irregular,the number of lamellar bodies in the plastids was decreased and was unevenly distributed.The pulmonary histopathological score in the high VT group was significantly higher than that in the spontaneous breathing group and the normal VT group (12.00 ± 2.00 vs.6.00 ± 1.51,8.50 ± 0.53,both P ＜ 0.01).③ Compared with spontaneous breathing group,IL-1β,IL-6,TNF-α,MPO,and MIP-2in serum and BALF in both mechanical ventilation groups were significantly increased,with more siguificant increase in the high VT group [serum IL-1 β (ng/L):104.2 ± 15.1 vs.20.3 ± 8.3,IL-6 (ng/L):46.6 ± 11.5 vs.22.7 ± 7.5,TNF-α (ng/L):39.4±6.5 vs.5.4± 1.9,MPO (ng/L):0.66±0.24 vs.0.06±0.03,MIP-2 (ng/L):109.2±25.8 vs.22.8±8.4;BALF IL-1 β (ng/L):121.5 ± 25.6 vs.24.0 ± 7.5,IL-6 (ng/L):136.7 ± 32.7 vs.31.4 ± 10.5,TNF-α (ng/L):98.0 ± 14.8vs.10.1 ±2.6,MPO (ng/L):0.80±0.31 vs.0.08±0.04,MIP-2 (ng/L):144.4±28.9 vs.41.2±20.7;all P ＜ 0.01].④ There were only a few p-ERK,Rac1 and F-actin positive expressions in the spontaneous breathing group.The positive expressions in normal VT group were increased.In high VT group,the positive expression of p-ERK was significantly increased;Rac1 and F-actin were mainly distributed in the cell membrane and cytoplasm respectively,the positive expressions were further enhanced.⑤ The gene expressions of ERK and Rac1,and protein expressions of p-ERK,Rac1 and F-actin in the high VT group were significantly higher than those in the spontaneous breathing group and normal VT group [ERK mRNA (2-△△Ct):8.23±2.83 vs.1,3.02± 1.38,p-ERK protein (gray value):1.15±0.36 vs.0.61 ±0.23,0.88±0.22;Rac1 mRNA (2-△△Ct):4.45 ±2.26 vs.1,1.22±0.39,Rac1 protein (gray value):0.91 ±0.16 vs.0.48±0.11,0.55 ± 0.10;F-actin protein (gray value):0.70± 0.09 vs.0.49 ± 0.08,0.55 ± 0.04;all P ＜ 0.01].Conclusion F-actin expression in lung tissue was up-regulated in rats with VILI,which resulted in reconstruction of AEC Ⅱ cyto-skeleton,and variation of cell membrane permeability through Rac 1 /MAPK/ERK sigualing pathway during VILI.

Objective To explore the clinical effect of high intensity focused ultrasound(HIFU)combined with transcatheter arterial chemoembolization(TACE)in the treatment of primary liver cancer.Methods A total of 106 patients with primary liver cancer were divided into two groups:50 cases were treated with TACE,and the other 56 were treated with combination of HIFU and TACE.The changes of AFP levels and the size of tumors after three months treatment were analyzed and compared with each other.The survival rates for one,two and three years were calculated with Kaplan-Meier method and compared between the two groups.Results In the two groups,AFP decreased significantly after treatment,but the combined group was superior to the other in AFP decrease.In the combined group,the 1-,2-and 3-year survival rates were higher than those in the TACE group with 82.3%,60.8%and 39.2% vs 68.0%,42.6%and 21.0%respectively(P＜0.01).No serious complications were seen,such as burn of skin,bleeding,gastrointestinal perforation. Conclusion The use of HIFU combined with TACE in the treatment of patients with primary liver cancers is feasible and safe.The combined group is superior to simple TACE for the management of primary liver cancers,and the former is more effective in decreasing AFP level and improving survival rates.