Objective:To explore the technical advantages of MR cholangiopancreatography (MRCP) with single breath holding high parallel acquisition factor 3-D variable flip angle fast spin echo (3D-SPACE) sequence.Methods:From November 2018 to March 2019, 75 patients who underwent MRCP examination in our hospital were prospectively enrolled, with single breath holding high parallel acquisition factor 3D-SPACE sequence and free breathing navigation gated 3D-SPACE sequence. Three experienced radiologists scored the overall image quality, artifacts, CBD visibility, left and right hepatic ducts, right anterior and posterior branches, second and third branches, main pancreatic duct and gallbladder duct with four scales. Paired t test was used for statistical analysis. Results:The scanning time of single breath holding method (18 s) was significantly shorter than that of free breathing diaphragm navigation method［264（226，313）s］, and the difference between the two methods was statistically significant ( Z=-7.520， P<0.001). The SNR, CR and CNR (8.31±4.23, 0.92±0.30, 11.46±5.77) of single breath holding method were lower than those of free breathing diaphragm navigation method (11.23±5.70, 0.93±0.38, 15.06±7.37), and the differences between the two methods were also statistically significant ( t=4.378, 3.429, 4.063, P<0.05). The overall image quality, artifact, the CBD, left and right hepatic duct, right anterior and posterior branchs, the second and third branches, main pancreatic duct and cystic duct of single breath holding method were higher than those of free breathing diaphragm navigation method, and the differences between the two methods were statistically significant ( P<0.001). Conclusions:Compared with the free breathing diaphragm navigation gated 3D-SPACE MRCP imaging method, the single breath holding high parallel acquisition factor 3D-SPACE MRCP imaging method has less artifacts and examination time, but higher visibility to pancreaticobiliary tree and work efficiency, which is worthy of further promotion.

Objective: The effect of total flavonoids of litchi (TFL) on nuclear translocation of nuclear factor-kappa B (NF- kappa B) in rat hepatic stellate cell line (HSC-T6) induced by transforming growth factor - beta 1 (TGF- beta 1) in vitro was studied to explore the mechanism of action of anti-hepatic fibrosis drugs. Methods: HSC-T6 was cultured in vitro, induced by TGFβ1 for 24 h, and then treated with TFL at 125, 250 and 500 μg/ml for 48 h. The effect of TFL on NF-κB nuclear translocation in HSC-T6 was observed by confocal laser microscopy. The effects of TFL on the expression of TLR4, p-IκB ɑ, p-NF-κB p65, NF-κB and Collagen I protein were detected by western blot. The expressions of TLR4 and p-NF-κB p65 were detected by immunofluorescence. Data were presented as mean±SEM. Homogeneity test of variance was performed and then followed by one-way analysis of variance (ANOVA). The multiple comparisons between groups were performed by LSD test. P < 0.05 was considered statistically significant. Results: Confocal laser scanning microscopy showed TFL inhibited the nuclear translocation of NF-κB in activated HSC-T6 in a concentration-dependent manner and TFL down regulated the protein expression levels of TLR4, p-IκB ɑ, p-NF-κB p65, NF-κB and collagen I protein in HSC-T6 in a concentration-dependent manner. Conclusion The mechanism of TFL against hepatic fibrosis may be related to the inhibition of nuclear translocation of NF-κb in the activated HSC-T6 and the expression of TLR4, P-iκbɑ, P-nf-κb p65, NF-κb and collagen I protein in HSC-T6.

The increased levels of intracellular reactive oxygen species (ROS) in granulosa cells (GCs) may affect the pregnancy results in women with polycystic ovary syndrome (PCOS). In this study, we compared the in vitro fertilization and embryo transfer (IVF-ET) results of 22 patients with PCOS and 25 patients with tubal factor infertility and detected the ROS levels in the GCs of these two groups. Results showed that the PCOS group had significantly larger follicles on the administration day for human chorionic gonadotropin than the tubal factor group (P < 0.05); however, the number of retrieved oocytes was not significantly different between the two groups (P > 0.05). PCOS group had slightly lower fertilization, cleavage, grade I/II embryo, clinical pregnancy, and implantation rates and higher miscarriage rate than the tubal factor group (P > 0.05). We further found a significantly higher ROS level of GCs in the PCOS group than in the tubal factor group (P < 0.05). The increased ROS levels in GCs caused GC apoptosis, whereas NADPH oxidase 2 (NOX2) specific inhibitors (diphenyleneiodonium and apocynin) significantly reduced the ROS production in the PCOS group. In conclusion, the increased ROS expression levels in PCOS GCs greatly induced cell apoptosis, which further affected the oocyte quality and reduced the positive IVF-ET pregnancy results of women with PCOS. NADPH oxidase pathway may be involved in the mechanism of ROS production in GCs of women with PCOS.
Abortion, Spontaneous ; epidemiology ; Acetophenones ; therapeutic use ; Adult ; Apoptosis ; drug effects ; Embryo Transfer ; Female ; Fertilization in Vitro ; Granulosa Cells ; metabolism ; Humans ; NADPH Oxidases ; antagonists & inhibitors ; Onium Compounds ; therapeutic use ; Oocyte Retrieval ; Oxidative Stress ; Polycystic Ovary Syndrome ; drug therapy ; Pregnancy ; Pregnancy Rate ; Reactive Oxygen Species ; metabolism

Background and purpose: AMP-activated protein kinase (AMPK) plays an important role in the regulation of cell metabolism and energy balance and is associated with cell proliferation, survival and multiple signaling pathways. Recent reports found that AMPK is involved in tumor suppression and drug resistance. The aim of this study was to explore the effect of AMPK on the anti-tumor effect of adriamycin and underlying mechanism in breast cancer MCF-7/adr cells. Methods:The anti-proliferative effects of adriamycin was detected by methyl thiazolyl tetrazolium (MTT) assay in MCF-7/adr, MCF-7/adr-vector and MCF-7/adr-AMPKαcells. The cell morphology in each group was stained with the lfuorescent dye Hoechst 33528, and the effects on apoptosis induction were examined by lfow cytometry (FCM). The intracellular concentration of adriamycin was detected by lfuorescence assay. The resis-tance-and apoptosis-related proteins were analyzed by Western blot. Results:The growth of breast cancer MCF-7/adr cells was inhibited by adriamycin in a dose-and time-dependent manner. The IC50 values at 24 and 48 h were (36.8±2.1) and (28.8±1.3) μg/mL, respectively. AMPKαover-expression enhanced the cytotoxic effect of adriamycin in MCF-7/adr-AMPKαcells in a dose-and time-dependent manner. Its IC50 values at 24 and 48 h were (16.0±0.7) and (4.2±0.2) μg/mL, respectively. Fluorescent morphological assay showed that AMPKαoverexpression contributed to adriamycin induced apoptosis in MCF-7/adr-AMPKαcells. After treatment with 1.0 μg/mL adriamycin for 48 h, the apoptosis rates of MCF-7/adr, MCF-7/adr-vector and MCF-7/adr-AMPKα cells were (12.0±1.4)%, (12.7±1.6)% and (32.0±4.2)%, respectively, indicating that overexpression of AMPKα enhanced the adriamycin-induced apoptosis in MCF-7/adr cells. Fluorescence microplate assay showed that over expression of AMPKαsigniifcantly increased the intracellular accumulation of adriamycin, in a concentration dependent manner. Western blot analysis showed that, compared with MCF-7/adr and MCF-7/adr-vector cells, the expressions of Bax, caspase-3 and cleaved PARP proteins were increased. Meanwhile, Bcl-2 and P-gp protein expressions were decreased in MCF-7/adr-AMPKαcells. Furthermore, the release of cytochrome c from mitochondria into the cytosol was also observed in MCF-7/adr-AMPKαcells. Conclusion:AMP-Kαoverexpression can enhance the chemosensitivity of breast cancer MCF-7/adr cells to adriamycin through inhibiting the drug effux transporter and regulating the expression of apoptosis-related proteins.

Objective To investigate the effect of sepsis on vecuronium-induced inhibition of acetylcholine release in neuromuscular junction in rats.Methods Thirty-six adult male SPF SpragueDawley rats,aged 2-3 months,weighing 200-220 g,were randomly divided into 3 groups (n=12 each) using a random number table:control group (group C),sham operation group (group S) and sepsis group (group Sep).Sepsis was induced by cecum ligation and puncture (CLP) in rats anesthetized with intraperitoneal chloral hydrate 350 mg/kg.At 12 h after CLP,the sciatic nerve-pretibial muscle was prepared.Vecuronium was added to the culture medium with the final concentration of 0.08 μg/ml,and the sciatic nerve-pretibial muscle was incubated for 15 min.Before and after administration,evoked endplate potentials (EPPs) and miniature endplate potentiais (MEPPs) were recorded by using intracellular microelectrode.EPP/MEPP ratio was calculated.Results Compared to C and S groups,EPPs,MEPPs and EPP/MEPP ratio were significantly increased before and after administration in group Sep.EPPs,MEPPs and EPP/MEPP ratio were significantly lower after administration than before administration in the three groups.Conclusion Sepsis can promote acetylcholine release in neuromuscular junction,thus weakening vecuronium-induced inhibition of acetylcholine release in neuromuscular junction in rats.

Objective To investigate the effects of total flavonoids of litchi chinensis sonn (TFL) on cell proliferation and the molecular mechanism in rat hepatic stellate cells (HSC-T6) activated by growth factor-β1 (TGF-β1). Methods HSC-T6 cells were treated by 0.25%Trypsin-EDTA and then were digested into single cell suspension by DMEM (10%FBS included), which were mixed with TGF-β1 (5μg/L). (1) MTT method was used to detect the proliferation of HSC-T6 cells. Cells were cultured in 96-well plate and were treated by different concentrations of TFL including TGF-β1 group, the control group (5‰DMSO included), and different concentrations of TFL groups (80, 160, 320, 640 and 800 mg/L TFL). Each group has three wells. The absorbance (A) value was measured by enzyme standard meter at the 490 nm wavelength after 24 h, 48 h and 72 h treatment. The cell inhibitory rate was calculated. The subsequent experimental drug concentration and drug treatment time were determined according to half inhibitory concentration (IC50). (2) The expression levels of NF-κB andα-SMA mRNA were detected by PCR (for mRNA) and Western blot assay (for protein). Cells were cultured in the 10 cm culture dish and were divided into different TGF-β1 groups, including TGF-β1 group, the control group (5‰DMSO included), and different concentrations of TFL groups (125, 250 and 500 mg/L TFL). After 48 h, related indicators were measured. Results At the same treatment time point, with the increased concentrations of TFL, A values were gradually decreased, and the cell inhibitory rates were gradually increased. There were no significant differences in the expressions of NF-κB andα-SMA mRNA between TGF-β1 group and control group. And there were no significant differences in the expressions of NF-κB andα-SMA mRNA between TFL125 group, TGF-β1 group and control group. There was a gradually decrease in the expressions of NF-κB andα-SMA mRNA and protein with the increased concentrations of TFL. Conclusion TFL can inhibit TGF-β1-induced HSC-T6 cell proliferation, which is involved in the inhibited expressions of NF-κB andα-SMA to anti-fibrotic effects in liver fibrosis.

Objective] To observe the influence of YIGU oral liquid on the pain threshold and bone mineral density in ovariectomized rats. [Methods] Eighty-four Female SD rats were randomly divided into 4groups:YIGU oral liquid group, calcitonin group,YIGU oral liquid combined with calcitonin group, saline control group.The ovariectomized rats were confirmed the successful model of osteoporosis,rats in the YIGU oral liquid group were administered with YIGU oral liquid, rats in the calcitonin group were administered with calcitonin, rats in the YIGU oral liquid combined with calcitonin group were administered with YIGU oral liquid group and calcitonin,rats in the saline control group were administered with saline.To detect the pain threshold and bone mineral density at the 10th day, 30th day, 60th day and 90th day respectively. [Results]At the 10th, 30th, 60th and 90th day after treatment, the pain threshold of the YIGU oral liquid group, calcitonin group,YIGU oral liquid combined with calcitonin group compare to saline control group, there was significant statistical difference( P<0.05).At the 10th, 30th day after treatment,the bone mineral density of the YIGU oral liquid group, cal-citonin group,YIGU oral liquid combined with calcitonin group compared with saline control group, there was no significant statistical difference( P>0.05). At the 60th,90th day after treatment, there was significant statistical difference( P<0.05).[Conclusion] YIGU oral liquid can effectively enhance pain thresh-old and bone mineral density of the ovariectomized rats to inhibit the osteoporosis pain.

Embryos with a poor morphological score at cleavage stage are usually discarded because they are considered unsuitable for transfer and cryopreservation. This study examined the in vitro blastocyst development after extended culture of these embryos and the clinical outcomes after transfer of these blastocysts in warming cycles. A total of 597 blastocysts (24.7%) were obtained from 2421 embryos with low morphological scores after extended culture. One hundred and sixty blastocysts (6.6%) with optimal morphology were vitrified. Embryo utilization rate was increased from 30.8% to 32.6%. After warming, 61 out of 92 blastocysts (66.3%) survived and were transferred in 44 cycles. The clinical pregnancy rate and the implantation rate were 40.9% (18/44) and 32.8% (20/61) respectively. Thirteen healthy babies were born, and 5 pregnancies aborted spontaneously. Our study suggested that some blastocysts derived from embryos with a poor morphological score can be successfully vitrified and give rise to live births. Selection and vitrification of viable embryos after extended culture of embryos with a poor morphological score may constitute a proposal to avoid embryo wastage.
Adult ; Cryopreservation ; methods ; Embryo Culture Techniques ; methods ; Embryo Transfer ; methods ; statistics & numerical data ; Female ; Fertilization in Vitro ; methods ; statistics & numerical data ; Humans ; Infertility ; pathology ; therapy ; Middle Aged ; Pregnancy ; Pregnancy Outcome ; Vitrification ; Young Adult

Objective To study the effect of Kanglaite combined with comprehensive therapy on advanced non-small cell lung cancer.Methods Sixty-one patients with advanced non-small cell lung cancer were randomly divided into treatment group ( n=31 ) and control group ( n=30 ).Both groups were given comprehensive therapy.Treatment group were additionally treated with intravenous injection of 200 ml Kanglaite.Clinical efficacy,quality of life,pain relief and adverse reactions of the two groups were observed.Results ( 1 ) Quality of life was improved in 20 cases (64.5％ ),stabled in 8 cases (25.8％),declined in 3 cases ( 9.7％ ) of treatment group,and in the control group there were 9 cases ( 30.0％ ) improved,9 cases ( 30.0％ ) stabilized,12 cases (40.0％ ) declined respectively.Quality of life in treatment group was higher than in control group ( U=2.91,P＜0.01 ).( 2 ) Pain relief:the number of patients with complete remission,partial remission,no change,and progression were 5 cases ( 16.1％ ),16 cases ( 51.6 ％ ),16 cases (51.6％ ),6 cases (19.4％) and 4 cases (12.9％ ) in treatment group,and in control group they were 2 cases(6.7％ ),9 cases (30.0％ ),11 cases(36.7％ ) and 8 cases(26.7％ ) respectively.The effect of treatment on pain relief in treatment group was better than that in control group ( U=2.32,P＜0.05 ).(3) Clinical efficacy:in the treatment group there were 12 cases (38.7％) with partial remission,14 cases (45.2％) stabilized,and 5 cases (16.1％ ) progressed,and in control group the numbers were 8 cases (26.7％ ),8 cases (26.7％ ) and 14 cases (46.7％ ) respectively.The clinical efficacy in treatment group was better than that in the control group( U=2.04,P＜0.05).(4) There were significant difference on the change of white blood cell count and gastrointestinal reactions Ⅲ and Ⅳ degrees between treatment group and contrl group [22.6％ (7/31) vs.53.3％(16/30),x2=6.139 P＜0.05;19.4％ (6/31) vs.46.7％ (14/30),x2=5.161,P＜0.05].Conclusion Kanglaite injection combined with comprehensive therapy can improve clinical efficacy of therapy for advanced non-small cell lung cancer,reduce the toxic adverse reaction,protect immunity system and improve the quality of life of patients.

ObjectiveTo study microRNA (miRNA) expression and role of cell cycle regulation in decidualized endometrial stormal cells (ESC) in vitro.MethodsESC was induced decasualization in vitro and matched with non-decidualized cells as controls.The expression repertoire of miRNA was measured by microarray chip and was validated by real-time PCR.Flow cytometry was used to identify ESC cycle during decidual reaction in vitro and after miRNA222 inhibitor was transfected into it.Results (1) Between decidualized and undecidualized stromal cells,there were 49 miRNAs significantly different expression by microarray chip,including 16 miRNA up-regulation and 33 miRNA down-regulation.hsa-miR-27b,30c,143,101,181 b,29b,30d,507,23 a,222,221 exhibited significantly differential expression between decicualized and undecidualized stromal cells by real-time PCR (P ＜0.05).(2) After miRNA222 inhibitor (NC-FAM) transfection to decidual ESC,ESC were cultured by FBS medium for 24 hours,the rate of transfection was 70％.ESC were transfected with miRNA 222 inhibitor and cultured for 48 hours,the percentage of ESC at Sphase of (6.2 ± 0.7 ) ％ were significantly lower than ( 10.9 ± 0.8 ) ％ in control group ( P ＜ 0.05 ) ; the percentage of ESC at G0/G1 phase increased at transfection group [ (77.5 ± 1.3 ) ％ vs.(73.0 ± 1.6) ％ at control group ],but there was no significant difference (P ＞ 0.05 ).Decasualization ESC were transfected with miRNA 222 inhibitor and cultured for 48 h,the percentage of ESC at S-phase was ( 3.3 ± 0.6) ％ in transfection group,which were significantly lower than (7.8 ± 0.9 ) ％ in control group ( P ＜ 0.05 ).The percentage of ESC at G0/G1 phase was ( 80.7 ± 1.6 ) ％ in transfection group and ( 74.9 ± 1.1 ) ％.In control group,which did not reached statistical difference ( P ＞ 0.05).ConclusionmiRNA was involved in ESC decidual process in vitro by regulating cell cycle.