Objective To set up a novel clinical method for storage of oocytes by slowly freezing and successful pregnancy by intracytoplasmic sperm injection (ICSI). Methods The frozen oocytes (n=33) in metaphase Ⅱ (28),which were normal in morphology,were denuded of their cumulus-corona complex.The protective freezing solution contained 1,2-propanediol (1.5 mol/L) and sucrose (0.35 mol/L) and the process was slow for freezing and rapid for thawing.The recipients of these oocytes received hormone replacement therapy (HRT). Results A high survival rate of oocytes (32/33) was obtained when the sucrose concentration was 0.35 mol/L.ICSI induced a higher fertility rate (23/28),a good embryo cleavage rate (21/23) and a satisfactory embryo morphology for thawed oocytes.Embryo transfers were performed in 6 cycles/6 cases. Four cases got clinical pregnancy by demonstration of four gestation sacs on B ultrasonic examination 7 weeks after ICSI,1 fetal was first trimester spontaneous abortion,another 3 survival fetal were ongoing pregnancy and the first case gestation week is 17 weeks. Conclusions This study is the first successful application of human oocyte cryopreservation in China and this method can increase the survival rate of freezing oocytes.It’s easy to carry out,low in cost and high in the recovery rate of oocytes after thawing.

Objective To analyse the associated clinical findings of newborn infants with HIE after intrauterine asphyxia with normal Apgar scores at birth and follow-up the neurologic sequelae. Methods 30 newborn infants admitted for HIE in Jan.to Dec. 2002 were analysed,they had :(1)intra-uterine asphyxia with Apgar scores of 8～10 at birth,(2)with neurological symptoms on the first day after birth,(3)diseases of infection,genetics and metabolism,and deformity were ruled out,(4)with characteristic findings of HIE on cranial ultrasound examinations and CT scans. Results All of 30 newborn infants had signs of depression from the first day after birth,22 infants of them sometimes were irritable.17 cases (56.7%) with mild encephalopathy and 13 cases (43.3%) with moderate encephalopathy,none of severe case,compared with the infants who had HIE with birth asphyxia,there were no statistical difference. Less than 40% cases had elevated BUN and CKMB,hyponatremia,hypocalcemia,hypoglycemia,acidosis,etc The incidence of them were the same as the infants who had HIE with birth asphyxia. The findings of cranial ultrasound examinations were different between the acute and chronic hypoxic injury of intrauterine asphyxia Follow-up 28 infants at the mean age of (11.7?3.8) months. Except 1 case of cerebral palsy,all of them are normal intelligent infants. There were 4 cases of dilated lateral ventricle and 1 case of dilated 3rd ventricle at birth; 6 cases of dilated ventricles found in follow-up and 1 of them had cerebral palsy,all of those infants recovered at 12～19 months of age. Conclusion Newborn infants who had intrauterine asphyxia without birth asphyxia might suffer from HIE and cerebral palsy later. Cranial ultrasound examinations showed some of them had fetal brain damages. Therefore,intensive care of such infants,preventing and treating intrauterine asphyxia could decrease the morbidity of HIE.

Objective To investigate the expression of angiotensin-converting enzyme 2(ACE2) protein and mRNA in kidney of different age spontaneously hypertensive rats(SHR) and the role of ACE2 in the development of hypertension. Methods Male SHR(n=30) were allocated to 5 groups by age: 1)one-month-old group(S1); 2)two-month-old group(S2); 3)three-month-old group(S3); 4)six-month-old group(S6); 5)nine-month-old group(S9). Sex and age matched Wistar-Kyoto normotensive rats(WKY)served as controls. The expression of ACE2 mRNA in kidney was determined by reverse transcription polymerase chain reaction(RT-PCR). The expression of ACE2 protein in kidney was assessed by immunohistochemistry and computer image analysis. Results 1)An age-dependent increase of SBP was observed in SHR before six months(P0.05). 2)An age-dependent increase of the ACE2 protein and mRNA levels in kidney was observed in both SHR and WKY(P0.05). The ACE2 protein and mRNA levels in kidney were lower in SHR than those in the matched WKY at every corresponding time point(P

Objective To investigate the long-term programming effects on pubertal reproductive function by immunological challenge in early life. Methods Female Sprague-Dawley rats were administered by endotoxin (lipopolysaccharide, LPS) at a dosage of 50 μg/kg and saline intraperitoneally on postnatal day 3 and 5. Body weight was measured weekly. Puberty onset ( vaginal opening) and oestrous cyclicity were monitored from postnatal day 30. At the age of 6 weeks, bilateral ovariectomy was performed. The histological and morphological change of the ovaries (the thickness of the theca interna and the number of different kinds of follicles) were observed and the immunoreactivity of the ovarian sympathetic nerve markers (low affinity receptor of nerve growth factor, p75NGFR) was evaluated by immune staining. Results Immunological challenge (exposed to LPS) in early life delayed vaginal opening significantly [LPS-treated (40.6 ±0.7) days versus controls ( 38. 6 ± 0. 5 ) days, P ＜ 0. 05], decreased the percentage of normal oestrous cyclicity ( LPS-treated 26. 1% versus controls 66. 8% , P＜ 0. 05 ) , decreased the total number of different types of follicles (primordial follicles: LPS-treated 610 ±47 versus controls 1181 ±57, P ＜ 0. 05; primary follicles: LPS-treated 624 ±41 versus controls 960 ± 30, P ＜ 0. 05 ; preantral follicles: LPS-treated 183 ± 16 versus controls 260 ± 14, P ＜ 0. 05; antral follicles: LPS-treated 32 ± 4 versus controls 79 ± 7, P ＜ 0. 05) and increased the thickness of the theca interna [LPS-treated ( 15. 8 ±0. 4) μm versus controls (11.4 ±0. 3) μm, P ＜ 0. 05]. The immunostaining of p75NGFR was obviously enhanced in the LPS-treated ovaries when compared with that of controls ( P ＜ 0. 05 ) . Conclusions Immunological stress during early critical developmental windows could have long dysfunctional effects on the pubertal reproductive function. It delayed puberty onset, reduced the percentage of the normal oestrous cycles, decreased follicles reserve and increased the thickness of the theca interna which might involve the up-regulation of the local ovarian sympathetic nerve activity.

Objective To screen an optimum method for in vitro culture of Helicobacter pylori-spe-cific CD4+T cells and apply it to immunodominant Th epitopes screening .Methods PBMCs were isolated from subjects positive for Helicobacter pylori infection and were stimulated with HpaA recombinant protein . Various induction conditions including serum containing mediums , concentrations of antigen and time were screened to obtain an optimum method for in vitro culture of Helicobacter pylori-specific CD4+T cells.The cells were harvested and stimulated using HpaA synthesized overlapping peptide pool .The percentage of an-tigen-specific CD4+T cells was evaluated by intercellular cytokine staining of interferon-γand the results were compared under different conditions .The possible immunodominant Th epitopes were screened by using synthetic overlapping peptides .Results Antigen-specific CD4+T cells were well cultured in RPMI 1640 culture medium containing human AB serum in comparison with those cultured in fetal bovine serum based medium.The highest percentage of antigen-specific CD4+T cells was achieved when stimulated with HpaA recombinant protein at the concentration of 0.2 μmol/L.CD4+T cells in response to the stimulation of 0.2μmol/L of HpaA recombinant protein was observed on the ninth day after culture and its peak was reached on the fifteenth day .A possible immunodominant Th epitope ( HpaA220-237 ) was screened in subjects with He-licobacter pylori-infection by using synthetic overlapping peptides .Conclusion Helicobacter pylori-specific CD4+T cells were successfully cultured in vitro by using RPMI 1640 culture medium containing human AB serum and stimulated with 0.2 μmol/L of HpaA recombinant protein for fifteen consecutive days .This cul-ture method could be applied to immunodominant Th epitopes screening and provide evidences for further in -vestigation on the development of Helicobacter pylori epitope-based vaccine .

Objective To investigate the role of acid-sensing ion channels (ASICs) in global cerebral ischemia-reperfusion (I/R) injury in rats. Methods Forty-eight adult male Sprague-Dawley rats weighing 250-310 g were randomly divided into 4 groups ( n = 12 each): sham operation group (group S); global cerebral I/R group (group I/R); normal saline group (group NS) and specific ASIC blocker amiloride group (group A). Global cerebral I/R was produced by occlusion of 3 vessels ( 10 min occlusion of the bilateral common carotid arteries and basilar artery) followed by reperfusion. In group NS and A, NS 6 ml/kg and amiloride 0.6 mg/kg were injected through femoral vein immediately before reperfusion respectively. Six rats in each group were selected, the dialysate in CA1 area was collected before ischemia (baseline), immediately after ischemia and during 20 min reperfusion (once every 10 min) for determination of lactate concentrations. The left 6 rats in each group were elected at 8 h of reperfusion and the open field test and inclined plane test were peeformed to assess neurological behavior.The rats were then sacrificed and brain tissues taken for microscopic examination and brain water content was calculated. Results Compared with group S, the concentration of lactate in the dialysate and brain water content were significantly increased and neurological deficits developed in group I/R and NS (P ＜ 0.05). Compared with group I/R, the concentration of lactate in the dialysate and brain water content were significantly decreased and neurological deficits were improved in group A ( P ＜ 0.05 ), but no significant change in the parameters mentioned above was found in group NS ( P ＞ 0.05). Microscopic examination showed that the damage to the brain tissues was attenuated in group A compared with group I/R. Conclusion ASICs are involved in the development of global cerebral I/R injury in rats.

Objective To investigate the pathogenic constitution of thyroid nodular diasease and it' s di-agnosis and treatment.Methods A retrospective analysis was performed on data of 576 patients accepted hospi-tal therapy with thyroid nodular diasease from January 2004 to June 2008.Results Definite diagnosis of all easeswere made by surgery biopsy or fine needle aspiration-cytology.529 eases were benign lesions, another 47 eases were malignant lesions.Condusions Ultra-sound , FNAC and frozen sections have important value in differenti-al diagnosis for the thyroid nodules, nodular goiter is most common, and which have a close relationship with thy-roid carcinoma.Regular surgical resection would reduce the recurrence of thyroid nodular diasease and avoid missed diagnosis for thyroid carcinoma.

Objective To summarize the experience of how to preserve parathyroid glands and their function in thyroidectomy. Methods Data of 872 patients with thyroid carcinoma who entered our hospital from May. 2006 to Oct. 2009 were retrospectively analyzed. Results Parathyroid damage rate was 2. 06% (18/872)and permanent hypoparathyroidism was 0. 23% (2/872). All occurred in total thyroidectomy. Conclusions Preserving blood supply for parathyroid and avoiding direct contusion or misresection are important for preventing hypoparathyroidism in thyroidectomy. Calcium supplementation should be given to patients with obviously low serum calcium level after surgery to prevent permanent hypoparathroidism.

Objective To investigate the effect of dezocine on emergence agitation (EA) during recovery from sevoflurane-based anesthesia in children.Methods Ninety ASA Ⅰ or Ⅱ children,aged 3-6 yr,weighing 14-31 kg,were randomized into 3 groups (n =30 each):fentanyl group (group F),dezocine group (group D)and control group (group C).Anesthesia was induced with inhalation of 8 ％ sevoflurane and iv injection of atracurium 0.5 mg/kg and remifentanil 1 μg/kg and maintained with inhalation of 2％-3 ％ sevoflurane and infusion of remifentanil at 0.1-1.0 μg· kg-1 · min-1.In groups F,D and C,fentanyl 2 μg/kg,dezocine 0.05 mg/kg and the equal volume of normal saline were injected intravenously 10 min before the end of surgery,respectively,and inhalation of sevoflurane was stopped at the end of operation.The extubation time and length of stay in the postanesthesia care unit (PACU) were recorded.Agitation and the duration were recorded when the children were in PACU.The degree of EA was assessed using the Pediatric Anesthesia Emergence Delirium (PAED) scale.Pain was evaluated using the Children' s Hospital of Eastern Ontario Pain Scale (CHEOPS) and sedation was measured with Ramsay score.The complications such as respiratory depression,postoperative nausea and vomiting were recorded within 24 h after operation.Results Compared with group C,the incidence of EA and PAED scores were significantly decreased and the duration of EA was shortened in groups F and D,CHEOPS scores were decreased at each time point after operation and Ramsay scores were increased at T1 in group F,and CHEOPS scores were decreased and Ramsay scores were increased at each time point after operation in group D (P ＜ 0.05).Compared with group F,the incidence of EA and PAED scores were significantly decreased,CHEOPS scores were decreased at each time point after operation,and Ramsay scores were increased at T2 (P ＜ 0.05),and no significant change was found in the duration of EA in group D (P ＞ 0.05).There was no significant difference in the extubation time and length of stay in the PACU between the three groups (P ＞ 0.05).Conclusion Dezocine 0.05 mg/kg can safely and effectively prevent EA during recovery from sevoflurane-based anesthesia in children and produces better efficacy than fentanyl.