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.

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.

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 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.

Objective To observe the expression of STEAP4 gene(a novel obesity-related gene) during the period of human preadipocyte differentiation and to explore the relationship between the STEAP4 gene expression and adipocytes differentiation,adipogenesis.Methods Human preadipocytes were cultured and differentiated into the matured adipocytes in vitro.Adipocytes morphology and lipid accumulation were observed during this process.Total RNA was extracted from adipocytes at various time points (preadipocyte,Day 0,Day 4,Day 6,Day 8,Day 11,Day 14,and Day 17) and the level of STEAP4 mRNA expression was measured by fluorescent real-time quantitative reverse transcriptase-polyme-rase chain reaction(RT-PCR).Results The level of STEAP4 mRNA expression remained high in preadipocytes.In the presence of differentiation medium (Day 4),there was a transient upregulation in the expression of STEAP4 gene.After that,with the human preadipocytes being differentiated into matured adipocytes,the expression of STEAP4 mRNA was downregulated and reached the lowest level in fully differentiated adipocytes.There was a significant difference between any 2 detected phases in the level of STEAP4 mRNA expression (Pa

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.

OBJECTIVETo explore the better therapy in the treatment of ganglion.

METHODSNinety cases of ganglion were randomized into a two-way quintuple puncture group, a common quintuple puncture group and a fire needling group, 30 cases in each one. In the two-way quintuple puncture group, the "9-in-1" multiple penetrating needling technique was used. In the common quintuple puncture group, the traditional "5-in-1" multiple penetrating needling technique was applied. In the fire needling group, the traditional multiple fire needling technique was adopted. The treatment was given once a day, 3 treatments made one session and the efficacy was analyzed statistically after 1 session treatment in the three groups.

RESULTSAll of the three therapeutic methods achieved the efficacy on ganglion. The curative rate was 96. 7% (29/30) in the two-way quintuple puncture group, which was better obviously than 66.7% (20/30) in the common quintuple puncture group and 60. 0% (18/30) in the fire needling group (both P<0. 01).

CONCLUSIONThe two-way quintuple puncture technique achieves the remarkably superior efficacy on ganglion as compared with the common quintuple puncture technique and fire needling technique.

Acupuncture Therapy ; Adolescent ; Adult ; Aged ; Female ; Ganglion Cysts ; therapy ; Humans ; Male ; Middle Aged ; Treatment Outcome ; Young Adult