Objective To establish a satisfactory method of isolating,culturing and determining gut neural crest stem cells,which may provide theoretical basis for clinical application.Methods The guts of embryonic mice removed and dissociated were plated into serum-free DMEM/F12 medium.The mitogen-free DMEM/F12 medium supplementd with 10% fetal bovine serum was used to induce differentiation of GNCSCs.Neurospheres and their derivations were determined with immunocytochemical and immunofluorescent staning.Results Neurospheres were generated in the simplified serum-free medium.The staining results showed that enteric neurospheres were GNCSCs and could differentiate into neurons,glial cells and smooth muscle cells by serum-induction.Conclusion GNCSCs have the capacity of self-renewal and proliferation,and give rise to neurons,glial cells and myofibroblasts.

Objective To discuss the clinical manifestations, imaging data and DSA findings of lacunar infarction (LI). Methods One hundred and thirty-three patients, admitted to our hospital from May 2002 to April 2008, were chosen in our study; these patients with first onset as LI were confirmed by Head CT or MR; the clinical manifestations and imaging data were retrospectively analyzed; DSA was also performed on these patients and DSA findings were concluded. Results One hundred and thirty-three patients were clinically manifested as pure motor hemiplegia (PMH, n=42, 31.6%) and sensorimotor stroke (SMS, n=36, 27.1%). Two hundred and eighty-three lesions were noted by CT/MR examinations, including 78 locating at the endocyst (27.6%) and 121 locating at the corona radiate+greater oval center (91.0%). Forty-four patients were noted as having 101 intracranial vessel lesions by DSA, including 38 patients with angiostenosis, 6 with Moyamoya and 1 with single intracranial aneurysm; of the patients with angiostenosis, 95 lesions (34 in the offending vessels and 61 in other vessels) were found. Among the DSA (+) patients, PMH (n=21) and SMS (n=10) were mainly noted with their lesions locating at the endocyst (n=23) and the corona radiate+greater oval center (n=31); At least 1 high-risk factor such as hypertension, diabete, hyperlipemia, coronary heart disease and arial fibrillation was found in 44 patients. Conclusion The pathogeneses of LI are various. Main artery infarction may co-exist in some cases. PMH and SMS are common with their lesions frequently locating at basal ganglia area and corona radiate of the cerebral hemisphere. High risk factor exists in most patients with cerebrovascular diseases.

OBJECTIVETo explore effective pretreated methods for hydrosalpinx before frozen embryo transfer (FET).

METHODSA randomized controlled study was performed on 229 FET cycles of hydrosalpinx patients. They were assigned to two groups by random digit table, Group A (94 cases), Group B (89 cases), and Group C (46 cases). Patients in Group A received transvaginal aspiration of hydrosalpinx combined with auricular point sticking. Those in Group B received transvaginal aspiration of hydrosalpinx group. Those in Group C received no transvaginal aspiration of hydrosalpinx. Pregnancy outcomes of FET, endometrial and subendometrial blood flow distribution on the embryo transfer day were compared among the three groups.

RESULTSThere was no statistical difference in the endometrial thickness on FET day, the number of transfer embryos, the number of transferred good quality embryos among the three groups (P > 0.05). The clinical pregnancy rate and the embryo implantation rate were significantly higher in Group A than in Group C (P < 0.05), and the clinical pregnancy rate was significantly higher in Group A than in Group B (P < 0.05). The early abortion rate and the transfer cycle cancel rate were significantly lower in Group A than in Group C (P < 0.05). Type A endometrial and subendometrial blood flow distribution was dominant in Group A, which was significantly higher in Group A than the rest two groups (P < 0.05). Type A distribution rate was also significantly higher in Group B than in Group C (P < 0.05).

CONCLUSIONTransvaginal aspiration of hydrosalpinx combined with auricular point sticking before FET could improve the endometrial receptivity and improve outcomes of IVF.

Embryo Implantation ; Embryo Transfer ; methods ; Female ; Fertilization in Vitro ; methods ; Humans ; Pregnancy ; Pregnancy Outcome ; Pregnancy Rate

Objective To determine the impact of fluorine and aluminum,and both action combined on the number of rat osteoclasts and bone resorption cultured in vitro and to explore its mechanisms.Methods The osteoclasts and bone marrow stromal cells (BMSCs) isolated from long bone of new born rats were cultured,respectively,in TC199 medium (containing 10％ fetal bovine serum) with fluoride,aluminum and fluoride combined with aluminum.The osteoclasts were inoculated in 96-well culture plate and ivory slice,BMSCs in 6-well culture plate,and culture medium was changed after 2 hours incubation.The cells were divided into control group,fluoride group,aluminum group and fluoride combined with aluminum group; the doses of sodium fluoride were 0,1.0 × 10-4,0,1.0 × 10-4 mol/L and the doses of aluminum chloride were 0,0,1.0 × 10-5,1.0 × 10-5 mol/L,respectively.Tartrate-resistant acid phosphatase (TRAP) staining positive cells were counted under light microscope after TRAP staining on the 5th day and the pit formed in ivory slices were measured by histomorphometry after staining with toludine blue.The expression of osteoprotegerin(OPG) and receptor activator of nuclear factor kappa-B ligand (RANKL) was detected by real-time fluorescence quantitative PCR in BMSCs after 8 h treatment.Results ① Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the numbers of osteoclasts (F =7.15,6.56 and 7.98,respectively,all P ＜ 0.05).The numbers of osteoclasts in fluoride group,aluminum group and fluoride combined with aluminum group[(136.9 ± 22.99),(135.4 ± 23.5),(163.0 ± 24.4) per well] were higher than that in the control group[(92.5 ± 22.1) per well,all P ＜ 0.05].② Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the resorption pit area on ivory slices(F =10.47,12.64,14.29,respectively,all P ＜ 0.05).The resorption pit area on ivory slices in fluoride group,aluminum group and fluoride combined with aluminum group[(0.242 ± 0.031),(0.293 ± 0.026),(0.333 ± 0.016)mm2 per slice] was higher than that in the control group [(0.088 ± 0.030)mm2 per slice,all P ＜ 0.05].③Fluoride,aluminum and the interactive effects of fluoride and aluminum all had impact on the expression ratios of RANKL/OPG in BMSCs (F =8.15,15.38,23.59,respectively,all P ＜ 0.05).The expression ratios of RANKL/OPG in BMSCs in fluoride group,aluminum group and fluoride combined with aluminum group [(193.98 ± 137.93)％,(326.11 ± 176.78)％,(599.84 ± 275.82)％] were higher than that in the control group[(100.00 ± 56.02)％,all P ＜ 0.05].Conclusions Both fluoride and aluminum can cause increase in the number of osteoclasts in vitro and promote cell differentiation and bone resorption activity,which may be related to increased expression ratio of RANKL/OPG mRNA in BMSCs.The stimulating effects of fluoride on osteoclasts differentiation and bone resorption is enhanced by aluminum.

Objective To determine the effects of fluoride on osteoclasts's quantity and bone resorption function in vitro and its mechanisms. Methods The osteoclasts and bone marrow stromal cells(BMSCs) isolated from long bone of new born rats were cultured respectively in TC199 medium (containing 10% fetal bovine serum) with fluoride. The osteoclasts were inoculated in 96-well culture plate and ivory slice, BMSCs were inoculated in 6- well culture plate, respectively, medium were changed after 2 hours incubation. They were divided into control group, low-dose fluoride, medium-dose fluoride and high-dose fluoride groups, the doses of sodium fluoride were 0,2.5 × 10-5,5.0 × 10-5,10.0 × 10-5 mol/L, respectively. Tartrate-resistant acid phosphatase(TRAP) staining positive cells were counted under light microscope after TRAP staining on the 2nd and the 5th day and the pit formed in ivory slices were measured by histomorphometry after staining with toludine blue. The expression of receptor activator of NK-κβ ligand(RANKL) and osteoprotegerin(OPC) was detected by real-time fluorescence quantitative (337.5 ± 70.5), (447.5 ± 43.4), (472.9 ± 34.8), (475.3 ± 24.3)/well in the control group, the low-dose, mediumdose and high-dose fluoride groups, respectively. The differences were statistically significant between these groups and the control group (all P < 0.05). After in vitro culture for 5 days, the numbers of osteoclasts were (92.5 ± 22.1), (123.0 ± 26.4), (135.5 ± 22.2), (136.9 ± 23.0) per well in the control group, the low-dose, medium-dose and high-dose fluoride groups, respectively. The differences were statistically significant between these groups and the (0.088 ± 0.030), (0.100 ± 0.018), (0.152 ± 0.015), (0.242 ± 0.031 )mm2 per piece in the control group, the lowdose, medium-dose and high-dose fluoride groups, respectively. The values of medium-dose and high-dose fluoride BMSCs in the control group, the low-dose, medium-dose and high-dose fluoride groups were 100.00 ± 56.02, 144.95 ± 97.21,223.25 ± 184.48,193.98 ± 137.93, respectively. The values of medium-dose and high-dose fluoride groups were significantly higher than that of control group (all P < 0.05). Conclusions Fluoride can cause increase in the number of osteoclasts in vitro and promote their cell differentiation and bone resorption activity, which may be related to increased expression ratio of RANKL/OPG mRNA in BMSCs.

To determine the optimum conditions of supercritical CO2 extraction of Xiaoyaosan, and establish its fingerprint by gas chromatography-mass spectrometry (GC-MS), the yield of extract were investigated, an orthogonal test was used to quantify the effects of extraction temperature, pressure, CO2 flow rate and time, and fingerprint analysis of different batches of extracts were by GC-MS. The optimal extraction conditions were determined as follows: extraction pressure 20 MPa, extraction temperature 50 degrees C, CO2 flow rate 25 kg x h(-1), extraction time 3 h, and average yield 2.2%. The GC-MS fingerprint was established and 27 common peaks were found, whose contents add up to 81.89% of the total peak area. Among them, 21 compounds were identified, accounting for 53.20% of the total extract. The extraction process is reasonable and favorable for industrial production. The GC-MS method is accurate, reliable, reproducible, and can be used for quality control of supercritical CO2 extract from Xiaoyaosan.
Carbon Dioxide ; chemistry ; Chromatography, Supercritical Fluid ; methods ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Gas Chromatography-Mass Spectrometry ; methods

Objective To investigate the relationship between CYP19 gene polymorphism and Alzheimer disease. Methods Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) was used to analyze the allele frequency distribution at the Mfe Ⅰ site of CYP19 gene in 102 patients with Alzheimer disease and 121 healthy control subjects. Results The frequencies of CYP19 ml and m2 alleles showed significant differences between the patient group and the control group (66.2% vs 81.0% and 33.8% vs 19.0%, respectively, X2=12.696, P<0.05). In patients with Alzheimer disease, the frequencies ofm1/m1, m1/m2, m2/m2 genotypes of CYP19 gene were 44.1%, 44.1%, and 11.8%, respectively, showing significant differences from the frequencies in the control subjects (65.3%, 31.4%, and 3.3%, respectively, X2=12.384, P<0.05). Conclusion CYP19 gene polymorphism at the Mfe Ⅰ site is associated with the genetic susceptibility to Alzheimer disease.

The effect of high-frequency repetitive transcranial magnetic stimulation (rTMS) on potassium-chloride cotransporter-2 (KCC2) protein expression following spinal cord injury (SCI) and the action mechanism were investigated.SCI models were established in SD rats.Five groups were set up randomly:normal control group,SCI 7-day (7D) model group,SCI 14-day (14D) model group,SCI-7D rTMS group and SCI-14D rTMS group (n=5 each).The rats in SCI rTMS groups were treated with 10 Hz rTMS from 8th day and 15th day after SCI respectively,once every day,5 days every week,a total of 4 weeks.After the model establishment,motor recovery and spasticity alleviation were evaluated with BBB scale once a week till the end of treatment.Finally,different parts of tissues were dissected out for detection of variations of KCC2 protein using Western blotting and polymerase chain reaction (PCR) technique.The results showed that the BBS scores after treatment were significantly higher in SCI-7D rTMS group than in SCI-14D rTMS group (P＜0.05).As compared with normal control groups,The KCC2 protein in SCI model groups was down-regulated after SCI,and the decrease was much more significant in SCI-14D model group than in SCI-7D group (P＜0.05).As compared with SCI model groups,KCC2 protein in rTMS groups was up-regulated after the treatment (P＜0.05).The up-regulation of KCC2 protein content and expression was more obvious in SCI-7D rTMS group than in SCI-14D rTMS group (P＜0.05).It was concluded that 10 Hz rTMS can alleviate spasticity in rats with SCI,which might be attributed to the up-regulation of KCC2 protein.It was also suggested that the high-frequency rTMS treatment after SCI at early stage might achieve more satisfactory curative effectiveness.

Objective To analyze the awareness on health among high school students and its influencing factors in Guangdong. Methods Multi-stage sampling and questionnairs "2009 health awareness survey of the Chinese citizens" developed by our Department of Health, were used. Data were analyzed by multivariate multilevel model under MLwinN 2.19 software. Results The mean scores on knowledge and ideas, behaviors and related skills among 1606 high school students of Guangdong province, were 69.08 ± 14.81,60.05 ± 16.85 and 74.99 ± 21.17 respectively. Three items on health showed that they all related to each other and relations between grades (0.972,0.715 and 0.855) were greater than the individuals (0.565,0.426 and 0.438). Conclusion Factors as students from outside the Pearl River Delta region or from the rural areas, being male, at general secondary schools, at grade one, with poor academic performance and more pocket money etc., had lower levels on those related information of health.

Objective To investigate the intelligence standard for diagnose the sub-cretin children and children with mental retardation of socio-cultural type.Methods The full intelligence quotient(IQ),verbal intelligence quotient(VIQ)and performance intelligence quotient(PIQ)was tested by Wechsler scale(C-WISC)for mild iodine deficiency disordem children,children living in abnormal socio-cultural condition and normal children aged 7～14 years old in Qinba mountain area.The test results had been compared between the groups.Results There were no significant difference between psychomotor functioning well children and children living normal sociocuhural condition in VIQ,PIQ and full IQ(89.24±18.44 vs 90.75±17.58,87.58±15.78 vs 88.95±15.56,87.42±17.84 vs 89.02±17.18,t=1.14,1.19 and 1.24,respectively,all P＞O.05).PIQ and full IQ were significantly lower in mild iodine deficiency disorders children than in children with abnormal socio-cultural background (65.81±10.22 vs 72.33±13.23,62.42±12.31 vs 68.13±14.54,t=3.26,2.55,P＜0.01 or＜0.05,respectively).But the VIQ was not significantly different between these two groups.The average difference of VIQ and PIQ among mild iodine deficiency disorders children wag-0.32 without significant difierence(t=0.28,P＞0.05),however it was-2.91 among children under abnormal socio-cultural condition with significant difierenee(t=-3.59,P＜0.01).Conclusions IQ for iodine deficiency disorders children is characterized by that VIQ is damaged in parallel with PIQ,while that in children under abnormal soeio-cuhural condition is marked by that VIQ is retarded more severely than PIQ,which ean be used as an intelligence standard for differentiating the sub-cretin children from children wjth socio-cuhural mental retardation.