Brain ; metabolism ; pathology ; Child, Preschool ; Chromatography, High Pressure Liquid ; Diagnosis, Differential ; Folate Receptor 1 ; genetics ; metabolism ; Folic Acid ; blood ; cerebrospinal fluid ; metabolism ; Folic Acid Deficiency ; diagnosis ; drug therapy ; etiology ; Humans ; Infant ; Leucovorin ; therapeutic use ; Malnutrition ; complications ; diagnosis ; Tetrahydrofolates ; cerebrospinal fluid ; metabolism

Background and purpose:Ovarian cancer is associated with a high recurrence and mortality due to the existence of cancer stem cells. The purpose of this study was to investigate the effect of casticin (CAS) on the capability of self-renewal in ovarian cancer stem cell like cells (OCSLCs) derived from SKOV3 cell line. Methods:SKOV3 cell line cells were cultured in vitro, and OCSLCs were obtained and amplified through suspended culture with conditioned medium of the stem cells. The phosphorylation level of FoxO3a was analyzed using Western blot. The protein expression of FoxO3a was inhibited by FoxO3a speciifc siRNA transfection, and then the ratio of sphere-formation was detected. Results: Compared with parental cells, OCSLCs over-expressed phosphorylated FoxO3a (pFoxO3a) and had elevated ratio of sphere-formation [(3.1±0.3)% vs (34.8±6.8)%, P<0.05]. CAS significantly inhibited the capability of sphere-formation in OCSLCs and down-regulated the expression level of pFoxO3a. And the transfection of FoxO3a speciifc siRNA suppressed the protein expression of FoxO3a and attenuated the inhibitory effect of CAS on the sphere-formation of OCSLCs. Conclusion: Reduced expression level of pFoxO3a is involved in the effect that CAS inhibits sphere-formation of OCSLCs derived from SKOV3 cell line.

Objective To explore whether Forkhead O transcription factor-3a (FoxO3a) activity affects the capability of sphere-formation of ovarian cancer SKOV3 cell line.Methods Sphere-forming cells (SFCs) were obtained and amplified through suspended culture with conditioned medium of the stem cells in SKOV3 cell line.After SKOV3 cells were transfected with FoxO3a specific siRNA,the protein expressions of FoxO3a and Bmi1 and the ratio of sphere-formation were compared with Western blot and sphere-forming assay,respectively.Results Compared to parental cells,SFCs from SKOV3 cell line had higher ratio of sphere-formation and over-expressed Bmi1 and pFoxO3a.Transfection of FoxO3a specific siRNA down-regulated the protein expression of FoxO3a and upregulated expression of Bmi1 in SKOV3 cells,and enhanced the capability of sphere-formation.Conclusions Silence of FoxO3a leads to enhanced capability of sphere-formation in SKOV3 cell line.

Adolescent ; Adult ; Cause of Death ; Child ; Female ; Hematologic Neoplasms ; mortality ; surgery ; Hematopoietic Stem Cell Transplantation ; mortality ; Humans ; Male ; Middle Aged ; Tissue Donors ; Young Adult

AIM:To investigate the effect of cyclopamine , a Hedgehog ( Hh) signaling pathway inhibitor , on the biological behavior of intrahepatic cholangiocarcinoma cell line RBE .METHODS:The proliferation of RBE cells was detected by cell counting with Typan blue staining and MTT assay , and the apoptosis was analyzed by the flow cytometry . The Transwell invasive cabin assay was used to detect the invasion ability , and Western blot was used to determine the pro-tein expression of Gli 1 and MMP-9 in the RBE cells before and after cyclopamine treatment .RESULTS:Cyclopamine in-hibited the growth of RBE cells in a time-and dose-dependent manner .After cyclopamine treatment for 24 h, 48 h and 72 h, the apoptotic rates were significantly higher than those in control group .In control group , the number of cells invading through the Matrigel of invasion chamber was 154.52 ±13.61, while in experimental group it was 62.00 ±12.17, indica-ting that the invasion ability of the cells declined significantly .Furthermore , Western blot showed that the protein levels of Glil and MMP-9 in the RBE cells were decreased after treatment with cyclopamine for 24 h and 48 h.CONCLUSION:Blockage of the Hh signaling pathway with cyclopamine suppresses the proliferation , promotes the apoptosis and inhibits the invasion ability of RBE cells .

OBJECTIVETo investigate the effects of the plasma containing Qianlongtong Capsule (QLT)-containing plasma on the expression of the Smad4 gene in prostate stromal cells in vitro and provide some experimental evidence for the treatment of benign prostatic hyperplasia (BPH) with Chinese medicinal compound.

METHODSFifteen cases of BPH were equally randomized to three groups to be treated with QLT at a high dose (6 capsules once), a medium dose (3 capsules once), and a low dose (1.5 capsules once), tid, for 7 days consecutively. QLT-containing plasma was collected from the patients. Prostate stromal cells were identified by immunofluorescence when they became monolayered and cultured in the QLT-containing plasma for 24 hours, followed by detection of the expression of the Smad4 gene by real-time quantitative PCR and that of the Smad4 protein by Western blot.

RESULTSAfter treatment with the QLT-containing plasma, the expression of the Smad4 gene in the stromal cells was significantly increased in a dose-dependent manner as compared with the blank control and no-QLT groups (P < 0.01). The expression of the Smad4 protein was also markedly elevated after treatment. The differences were statistically significant between the blank control and medium-dose groups (P < 0.01), low-dose and medium-dose groups (P < 0.05), and high-dose and the other groups (P < 0.01), but not between the blank control and low-dose groups (P > 0.05).

CONCLUSIONQLT-containing plasma could inhibit the proliferation and improve the apoptotic index of prostate stromal cells in vitro, which was related to the elevation of the mRNA and protein expressions of Smad4.

Apoptosis ; Cells, Cultured ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Prostate ; drug effects ; metabolism ; RNA, Messenger ; genetics ; Smad4 Protein ; genetics ; metabolism ; Stromal Cells ; drug effects ; metabolism

Objective To investigate the distribution status of coal-burning fluorosis (endemic fluorosis) areas in Luoyang and to provide scientifc evidence for making strategies in prevention and control. Methods In 2006, a household per village was chosen to carry the general survey so as of disease condition, living habits and housing structure among 941 coal-burning pollution fluorine sickness natural villages in Yanshi, Mengjin, Xin'an, Luanchuan counties and Geely area which were under the jurisdiction of Luoyang. In the general survey, the sampled village having a population of more than 500 person was considered as a major survey village, and water fluoride, 8 - 12 year-old child fluorine spot on tooth and the urinary fluoride were surveyed. Water fluoride and the child urinary fluoride determination used the fluoride ion selective electrode method, and the children's dental fluorosis used Dean method. Results The endemic fluorosis of Luoyang existed 742 in endemic fluomsis villages, compared with history, a decrease of 199 in number. Ninety-six point seven per cent( 142 543/147 419) of the households were consuming smoke-free coal. Households using intact kitchens accounted for 93.6%( 137 919/147 419). Of which 63.0%(86 889/137 919) of kitchens were mixed up with bedrooms. Total 125 060 people were using coal- fired furnace for heating, of which 87.8%(109 802/125 060) had smoke-free facilities, 12.2%(15 258/125 060) had none. Among 52 endemic villages with population of more than 500 people surveyed, a total of 183 water samples were collected, 2 had water fluoride exceeding 1.0 rag/L, the highest water fluoride being 1.04 rag/L, averaging 0.39 mg/L Sixteen villages had a prevalence rate of dental fluorosis for children less than 30.00%, accounting for 30.8% (16/52), 36 endemic villages the prevalence of dental fluorosis detection rate of more than 30.00%, accounting for 69.2%(36/52). Twenty-thrce villages had a dental fluorosis index greater than 0.6, severe dental fluorosis was not found. Real-time measurement of 1408 urine samples of children aged 8 - 12 showed that urine fluoride highest value 6.88 nag/L, the minimum value of 0.10 mg/L, geometric mean 1.10 mg/L. The prevalence rate of dental fluorosis for children was 36.06%. Conclusions In Luoyang city, numbers of coal-burning endemic fluorosis villages are less than before, children's dental fluorosis has significantly declined, however some people still use kitchens connecting with bedrooms and lack smoke-free facilities, they need to be educated to change lifestyle and improve furnace to reduce soot fluoride pollution.

OBJECTIVETo explore the scientificity of Yunqi theory based on incidence of disease related recognitions such as year evolutive phase, host climatic qi, six climatic factors, celestial manager qi, qi in the earth by analyzing data of hypertension incidence.

METHODSA total of 1,265 inpatients with confirmed first diagnosis as hypertension were recruited at Dongzhimen Hospital, Beijing University of Chinese Medicine from February 4, 1999 to February 4, 2011. Each period (including year evolutive phase, host climatic qi, guest climatic qi) was deducted based on Yun-qi theory. Their ratios of different phases and qi in the total number of hypertension inpatients number were calculated and statistically tested.

RESULTSThere was statistical difference in ratios at year evolutive phase, celestial manager qi, and qi in the earth (P < 0.05).

CONCLUSIONThe incidence of hypertension has certain difference in each period, which is consistent with Yun-qi theory.

Beijing ; epidemiology ; Humans ; Hypertension ; epidemiology ; Incidence ; Medicine, Chinese Traditional ; Qi ; Research

Objective To investigate the correlation between 24 h urinary protein quantitation and pregnancy outcome in patients with pre-eclampsia.Methods A total of 332 pre-eclampsia patients were selected in Tongji Hospital from January 2014 to December 2016.The patients were divided into microalbuminuria group(24 h urinary protein quantification ＜ 0.3 g,n =46),mild proteinuria group (0.3 g ≤ 24 h urinary protein quantification ＜ 2.0 g,n =98),moderate proteinuria group (2.0 g ≤ 24 h urinary protein quantification ＜ 5.0 g,n =71) and severe proteinuria group(24 h urinary protein quantification ≥ 5.0 g,n =117) according to the results of 24 h urinary protein quantification.The pregnancy outcomes were compared between the four groups.Results The 24 h urinary protein quantification and the serum creatinine,urea nitrogen,uric acid levels in the mild proteinuria group,moderate proteinuria group and severe proteinuria group were significantly higher than those in the microalbuminuria group (P ＜ 0.05);and gestational week was significantly shorter than that in the microalbuminuria group (P ＜ 0.05).The 24 h urinary protein quantification and serum urea nitrogen,uric acid levels in the moderate proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and gestational week was significantly shorter than that in the mild proteinuria group (P ＜ 0.05);but there was no significant difference in serum creatinine level between the two groups (P ＞ 0.05).The 24 h urinary protein quantification,serum creatinine,urea nitrogen and uric acid levels in the severe proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and the gestational week was significantly lower than that in the mild albuminuria group (P ＜ 0.05).The 24 h urinary protein quantification in the severe proteinuria group was significantly higher than that in the moderate proteinuria group (P ＜ 0.05),but there was no significant difference in the gestational week and serum creatinine,urea nitrogen,uric acid levels between the two groups (P ＞0.05).There was no significant difference in the rates of cesarean section and spontaneous labor between the four groups (P ＞0.05).The rate of induced labor in the moderate proteinuria group and the severe proteinuria group was significantly higher than that in the mild albuminuria group and the microalbuminuria group (P ＜ 0.05).There was no significant difference in the rate of induced labor between the mild proteinuria group and the microalbuminuria group (P ＞ 0.05).There was no significant difference in the rate of induced labor between the severe proteinuria group and the moderate proteinuria group (P ＞ 0.05).The incidence of complications in microalbuminuria group,mild proteinuria group,moderate proteinuria group and severe proteinuria group was 30.43％ (14/46),47.96％ (47/98),74.65％ (53/71) and 74.36％ (87/117) respectively;the incidence of complications in the moderate proteinuria group and the severe proteinuria group was significantly higher than that in the microalbuminuria group and the mild albuminuria group (P ＜ 0.05),but there was no significant difference in the incidence of complications between microalbuminuria group and mild albuminuria group (P ＞ 0.05),there was no significant difference in the incidence of complications between the moderate proteinuria group and the severe proteinuria group (P ＞0.05).The incidences of premature birth and neonatal asphyxia in the mild proteinuria group were significantly higher than that in the microalbuminuria group (P ＜ 0.05),and the body mass of the neonates was significantly lower than that in the microalbuminuria group (P ＜0.05),but there was no significant difference in the perinatal mortality rate and the incidences of fetal growth restriction(FGR) and poor neonatal resuscitation between the two groups (P ＞ 0.05).The incidences of FGR,premature birth,neonatal asphyxia,poor neonatal resuscitation and the perinatal mortality in the moderate proteinuria group and severe proteinuria group were significantly higher than those in the microalbuminuria group (P ＜ 0.05);and neonatal body mass was significantly lower than that in the mieroalbuminuria group (P ＜ 0.05).The incidences of FGR,premature birth and poor neonatal resuscitation and perinatal mortality in the moderate proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and the neonatal body mass was significantly lower than that in the mild proteinuria group (P ＜ 0.05);but there was no significant difference in the neonatal asphyxia incidence between the two groups (P ＞ 0.05).The incidences of FGR,premature birth,neonatal asphyxia,poor neonatal resuscitation and perinatal mortality in the severe proteinuria group were significantly higher than those in the mild proteinuria group (P ＜ 0.05);and the body mass of the newborns was significantly lower than that in the mild albuminuria group (P ＜ 0.05).The incidence of neonatal asphyxia in the severe proteinuria group was significantly higher than that in the moderate proteinuria group (P ＜ 0.05),but there was no significant difference in the incidences of FGR,premature birth,poor neonatal resuscitation,perinatal mortality and neonatal body mass between the two groups (P ＞ 0.05).Conclusions The of 24 h urinary protein quantitation is closely related to the pregnancy outcome in patients with pre-eclampsia,the 24 h urinary protein quantification should be regularly detected in the patients with pre-eclampsia.When the urinary protein quantitation is more than 2.0 g,the incidences of maternal complications and poor prognosis of the perinatal infants is significantly higher,but the boundary value of the 24 h urinary protein quantitation for the diagnosis of severe pre-eclampsia still needs further large sample study.

[Objective] To investigate the expression of MICA mRNA and protein in osteosarcoma and give a more comprehensive understanding to its immune evasion.[Methods] RT-PCR was used to analyze MICA mRNA in 11 osteosarcoma tissues and 5 osteosarcoma cell lines.MICA expression was examined in 66 paraffin-embedded osteosarcoma tissues and 6 paraffin-embedded normal bone tissues by immunohistochemistry,and MICA protein in 9 fresh osteosarcoma tissues was detected by Western blot too.MICA surface expression was estimated by flow cytometry.[Results] Nine of eleven (81.8%) osteosarcoma specimens and all of the five cell lines consistently expressed MICA mRNA.Up-regulation of MICA expression was found in osteosarcoma (34/66,51.6%),compared with normal bone tissues (0/6,0%).The cell lines Saos-2,MG63,and HOS showed positive surface MICA expression,while the U2OS and OS732 showed very limited expression.[Conclusion] MICA mRNA and protein predominantly expressed on osteosarcoma tissues and cell lines.