Objective To investigate the effect of Oct4 gene silencing on apoptosis in human colorectal cancer SW480 cell lines and its possible mechanism. Methods SW480480 cells at the late logarithmic phase were divided into 3 groups. (1) control group (Con): no virus transfection; (2) negative control group (NC): transfected with negative control viruses; (3) RNA interference group (RNAi): transfected with Oct4-shRNA lentiviral vector plasmids. In the above three groups, the Oct4 mRNA level was detected by real-time quantitative PCR, the protein levels of Oct4 and p-Akt were detected by Western blotting, and the proportion of Oct4+ and CD44+ cells and apoptosis of the tumor were analyzed by flow cytometry. Results Compared with those in the NC group, the Oct4 mRNA levels (1.00 vs 0.19±0.02), the Oct4 protein levels (0.032±0.004 vs 0.007±0.001), the p-Akt protein levels (0.11±0.03 vs 0.03±0.01), the proportion of Oct4+ cells (91.53% vs 10.70%) and the proportion of CD44+ cells (59.69% vs 23.58%) in the RNAi group significantly reduced, while cells apoptosis in the RNAi group significantly increased (43.2%±4.5% vs 12.1%±1.8%, P<0.01). Conclusion Oct4 gene silencing could significantly reduce the proportion of Oct4+ and CD44+ cells, and obviously increase apoptosis in SW480 cells, which may be related to the PI3K/Akt pathway.

OBJECTIVETo study the effect of Guishen Pill (GSP) on expression levels of Oct-4, MVH, and Egr-1 in mice with diminished ovarian reserve (DOR).

METHODSTotally 40 female C57BL/6J mice were randomly divided into 4 groups, the normal control group, the model group, the GSP group, and the dehydroepiandrosterone (DHEA) group, 10 in each group. Pregnant mare serum gonadotropin (PMSG), human chorionic gonadotropin (HCG), and prostaglandin F2α (PGF2α) were sequentially administrated to produce superovulation. The DOR model was established by exposing to ozone inhalation. Mice in the GSP group were intragastrically administered with GSP at 0.3 mL. Those in the DHEA group were intragastrically administered with DHEA at 0.3 mL. Equal volume of normal saline was intragastrically administered to mice in the normal control group and the model group. All mice wer treated for 21 days. Serum levels of estrogen (E2), progestogen (P), and anti-Müllerian hormone (AMH) were measured by ELISA. Changes of Oct-4, anti-AMH, and early growth response gene-1 (Egr-1) mRNA in ovaries were dtected by Real-time PCR.

RESULTSCompared with the model group, serum levels of E2, P, and AMH, as well as contents of estrogen receptor (ER), progestogen receptor (PR), MVH, and Oct-4 mRNA significantly increased in the GSP group and the DHEA group (P < 0.05).

CONCLUSIONGSP could improve expression levels of Oct-4, MVH, and Egr-1 mRNA in DOR mice and their ovarian function.

Animals ; Anti-Mullerian Hormone ; metabolism ; Dehydroepiandrosterone ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Early Growth Response Protein 1 ; metabolism ; Estrogens ; Female ; Mice ; Mice, Inbred C57BL ; Octamer Transcription Factor-3 ; metabolism ; Ovarian Reserve ; Ovary ; Pregnancy ; Receptors, Estrogen ; metabolism ; Superovulation

Objective To investigate the cause of metastatic metatarsalgia after hallux valgus surgery and the clinical outcomes of Weil osteotomy for metastatic metatarsalgia after hallux valgus surgery. Methods From July 2009 to Janurary 2012, data of 27 patients (27 feet) with metastatic metatarsalgia of 2nd to 4th head of metatarsal bone after hallux valgus surgery who had been treated by Weil osteotomy were retrospectively analyzed. There were 1 male and 26 females with an average age of 51 years (range, 28-73 years). Metatarsalgia occurred 6-24 months after operation for hallux valgus. 13 feet underwent mini-invasive cervi-cal wedge osteotomy of the first metatarsal, 7 with Chevron procedure, 5 with Akin procedure, and 2 with Lapidus procedure. No shortening in first metatarsal was found in 5 feet with Akin osteotomy, while there were varying degrees of shortening in first meta-tarsal in the remaining 22 feet. The clinical results were evaluated by American Orthopaedic Foot and Ankle Society (AOFAS) and visual analogue scale (VAS) scores. The preoperative and postoperative plantar pressure changes were tested by plantar pressure tester. Results 25 Patients were followed up for 12-42 months (average, 24 months). Among these 25 cases, the metatarsalgia of 23 cases were completely disappeared. The metatarsal plantar lateral metastatic pain occurred in the remaining 2 cases (2 feet) and 1 was relieved by the foot pad, 1 was cured by re-Weil osteotomy. AOFAS score was 46.82 ± 6.13 before surgery and 90.63 ± 1.65 after surgery. The VAS score was 7.5 (6, 7) before surgery and 0.5 (0, 1.0) after surgery. The last follow-up, according to the score of AOFAS toe metatarsophalangeal-interphalangeal joint, of which 23 cases were excellent, 1 good, 1 poor;the excellent and good rate was 96%(24/25). Preoperative pressure under 2nd to 5th metatarsal head were 3.12±1.62 Pa, 5.81±1.92 Pa, 4.63± 2.10 Pa, 3.37±1.57 Pa, 1.67±1.20 Pa and postoperative were 3.33±1.35 Pa, 3.89±1.08 Pa, 3.65±1.96 Pa, 2.25±1.23, (1.48±1.11) Pa. Postoperative pressure under 2nd to 5th metatarsal head were significantly decreased. Conclusion Weil osteotomy can effec-tively adjust the length of the metatarsal and the height of metatarsal head, thus effectively improve the pressure under the metatar-sal head, so it could reach a good effect in the treatment of metastatic metatarsalgia after hallux valgus surgery.

Objective To investigate the effect of Oct4 gene silencing on apoptosis in human colorectal cancer SW480 cell lines and its possible mechanism. Methods SW480480 cells at the late logarithmic phase were divided into 3 groups. (1) control group (Con): no virus transfection; (2) negative control group (NC): transfected with negative control viruses; (3) RNA interference group (RNAi): transfected with Oct4-shRNA lentiviral vector plasmids. In the above three groups, the Oct4 mRNA level was detected by real-time quantitative PCR, the protein levels of Oct4 and p-Akt were detected by Western blotting, and the proportion of Oct4+ and CD44+ cells and apoptosis of the tumor were analyzed by flow cytometry. Results Compared with those in the NC group, the Oct4 mRNA levels (1.00 vs 0.19±0.02), the Oct4 protein levels (0.032±0.004 vs 0.007±0.001), the p-Akt protein levels (0.11±0.03 vs 0.03±0.01), the proportion of Oct4+ cells (91.53% vs 10.70%) and the proportion of CD44+ cells (59.69% vs 23.58%) in the RNAi group significantly reduced, while cells apoptosis in the RNAi group significantly increased (43.2%±4.5% vs 12.1%±1.8%, P<0.01). Conclusion Oct4 gene silencing could significantly reduce the proportion of Oct4+ and CD44+ cells, and obviously increase apoptosis in SW480 cells, which may be related to the PI3K/Akt pathway.

ObjectiveTo evaluate the curative effect difference between Tong's anterior mesh suspension and modified tension-free vaginal tape-obturator (TVT-O). Methods75 SUI cases were random divided into two groups: Group A (35 cases) were accepted Tong's anterior mesh suspension and group B (40cases) were accepted modified tension-free vaginal tape- obturator. Curative effects, operation safety,and complications and so on, were recorded and compared. ResultsThere had no statistical differences between these two groups on operation time, bleeding lose, operation injury, immediate postoperative urinary retention andcure rates, but the operation expense of group A was cheaper than group B [ (980. 74 ±212.45)yuan vs (2879.06 ±467. 13)yuan , P ＜0.05). ConclusionThe curative effect between two methods were similar. It's a little complicated and fit to be popularized in large hospital to modified tensionfree vaginal tape-obturator. Tong's anterior mesh suspension was performed under completely direct vision,not through pelvic cavity, and it was an economic, convenient and easy way, which not only fit to be popularized in basic hospital, but also a surgical remedial measure to the failures who accepted these operations,such as TVT, SPARC, IVS, MONARC, TOT, TVT-O, etc.

The effect and underlying mechanism of Bu-Shen-An-Tai recipe on ovarian apoptosis in mice with controlled ovarian hyperstimulation (COH) implantation dysfunction were studied.The COH implantation dysfunction model in mice was established by intraperitoneal injection of 7.5 IU pregnant mare's serum gonadotrophin (PMSG),followed by 7.5 IU human chorionic gonadotrophin (HCG) 48 h later.Then the female mice were mated with male at a ratio of 2:l in the same cage at 6:00 p.m.The female mice from normal group were injected intraperitoneally with normal saline and mated at the corresponding time.Day 1 of pregnancy was recorded by examining its vaginal smears at 8:00 a.m.of the next day.Fifty successfully pregnant mice were equally randomly divided into 5 groups:normal control pregnant group (NC),COH implantation dysfunction model group (COH),low dosage of Bu-Shen-An-Tai recipe group (LOW),middle dosage of Bu-Shen-An-Tai recipe group (MID) and high dosage of Bu-Shen-An-Tai recipe group (HIGH).Then from day 1,the mice in different groups were respectively intragastrically given corresponding treatments at 9:00 a.m.for 5 consecutive days.The concentrations of 17β-estradiol (E2) and progesterone (P4) were determined by radioimmunoassay (RIA).The ultrastructural changes of ovarian tissues were observed by transmission electron microscope (TEM).The histopathological changes of ovarian tissues were observed by HE staining.The number of atretic follicles and pregnant corpus luteum were also recorded.TUNEL was applied to measure apoptotic cells of ovarian tissues.Western blotting was used to detect the protein expression of apoptosis-related factors like Bax,Bcl-2 and cleaved-caspase-3 in ovarian tissue of mice.The results showed that ovarian weight,the concentrations of E2 and P4,the number of atretic follicles and pregnant corpus luteum,as well as the apoptosis of granulosa cells were significantly increased in the COH group.The ultrastructures of ovarian tissues in the COH group showed that chromatin in granulosa cells was increased,agglutinated,aggregated or crescent-shaped.The focal cavitation and the typical apoptotic bodies could be seen in granulosa cells in the late stage of apoptosis.After the treatment with different doses of Bu-Shen-An-Tai recipe,the ultrastructural changes of ovarian granulosa cells apoptosis were dramatically improved and even disappeared under TEM.Visible mitochondria and mitochondrial cristae were increased and vacuoles were significantly reduced.The lipid dropltes were shown in a circluar or oval shape.The protein expression levels of Bax and cleaved-caspase-3 were decreased,and the expression of Bcl-2 protein was increased after treatment.It was concluded that Bu-Shen-An-Tai recipe can inhibit the apoptosis of ovarian granulosa cells,probably by up-regulating the protein expression of Bcl-2 and down-regulating Bax and cleaved-caspase-3,which contributes to the formation and maintenance of ovarian corpus luteum.It's helpful to promote the embryonic implantation,to reduce embryo loss and ultimately to improve the success rate of pregnancy.

In this study, we investigated the pharmacokinetics parameters of SPIO-shRNA dual functional molecular probe and observed the main organ distribution by MRI in vivo. Eighteen New Zealand white rabbits were randomly divided into three groups and injected intravenously with different doses of SPIO-shRNA molecular probe, respectively. The blood samples were collected to analyze the pharmacokinetic parameters by measuring the iron content at 30 minutes before and after the injection. Twenty-four Kun Ming (KM) mice were randomly divided into 4 groups: the control group was injected intravenously with physiological saline 200 µL per mouse via the tail vein, the other 3 groups were injected intravenously with different doses of SPIO-shRNA molecular probe. MRI observation was performed in 24 hours, and the liver, spleen, kidney, brain and muscle were collected for iron quantification with Prussian blue staining to determine distribution of the SPIO-shRNA molecular probe in the main organ in vivo. Our results suggest that the molecular probe blood half-life is more than 3 hours. The data of MRI suggest the probe was distributed in liver and spleen, and the MRI signal was reduced with the increase in probe's doses (P < 0.05). The results of Prussian blue staining confirmed the results of MRI. Most of the probe could escape the phagocytosis of mononuclear phagocyte system. Our data provide the pharmacokinetic and distribution of SPIO-shRNA molecular probe in organs. Meanwhile, it suggests the choice of the time and dose of probe for MR imaging of tumor in vivo.
Animals ; Half-Life ; Magnetic Resonance Imaging ; Magnetite Nanoparticles ; Mice ; Molecular Probes ; pharmacokinetics ; RNA, Small Interfering ; chemistry ; Rabbits

Background c-Myc,EZH2 and p27 were defined to modulate the behavior of prostate cancer with pro-tumoral or anti-tumoral effects and had ability in predicting prostate cancer progression,but the research of their co-expression value of prognosis is rarely.This study aimed to investigate the prognostic value of combining tri-marker together in patients with intermediate-risk prostate cancer after surgery.Methods Expression levels of c-Myc,EZH2 and p27 in 129 patients with intermediate-risk prostate cancer were assessed using immunohistochemistry in a semi-quantitative manner.The expression profiles of these three markers were analyzed and investigated for association with biochemical recurrence.Results In all,fifty of 129 cases experienced biochemical recurrence during a median follow-up time of 31 months (range,6-60 months).Of these relapse patients,one case without and 10 cases with any single positive marker were observed; 39 cases were detected with any two or all three positive markers (22 cases with any two and 17 cases with all three positive markers).Survival analysis showed that patients with over-expression of c-Myc or EZH2,and lower expression of p27 manifested significantly higher biochemical recurrence rates.Subsequent multivariate analysis revealed that c-Myc,EZH2 and p27 expression statuses showed potential in predicting relapse,respectively.Notably,combining three markers together as a "composite index" (0 or 1,vs.2 or 3 positive markers) provided powerful prognostic value (HR 6.57,95％ CI 3.02-14.31,P ＜0.001).There was a significant difference between the patient subgroups with 0 or 1 and those with 2 or 3 positive markers expression statuses,and tri-marker composite index was an independent risk factor for predicting relapse in patients with intermediate-risk prostate cancer after surgery.Conclusion Composite index of c-Myc,EZH2,and p27 can be valued as powerful prognosis parameter for intermediate-risk prostate cancer patients after the surgery,and postoperative adjuvant therapy can be adopted accordingly.

OBJECTIVETo study the genotype frequencies of peroxisome proliferators-activated -receptors-gamma C161-->T gene and its possible association with the metabolic syndrome and dietary intakes.

METHODSThe PCR-PFLP method was used to detect the polymorphism of PPARgammaC161-->T gene of 224 adults with metabolic syndrome and 224 normal adults in Shanghai. Their physical examinations, dietary investigation and the serum biochemistry were analyzed.

RESULTS(1) The genotype frequencies of PPARgamma C161-->T CC, CT and TT were 32.4%, 49.6% and 18.0% respectively, which were in agreement with Hardy-Weinberg equilibrium. There was no significant difference in the distribution of genotypes or allele between the metabolic syndrome group and the control group, and the result was the same between male and female subjects. (2) The levels of body mass index,waist width and hip width were significantly different among three genotypes groups. Subjects of the CT genotype had the highest levels. (3) There was significant difference in the negative correlation with the intake of protein and serum TG levels in the metabolic syndrome group.

CONCLUSIONThe results suggested PPARgamma gene C161-->T should be associated with body mass index, waist width and hip width. It might contribute to the heterogeneity in diet response to TG.

Adolescent ; Adult ; Aged ; Alleles ; Causality ; China ; Diabetes Mellitus, Type 2 ; genetics ; Diet ; Female ; Gene Frequency ; Genotype ; Humans ; Male ; Metabolic Syndrome ; genetics ; Middle Aged ; Obesity ; genetics ; PPAR gamma ; genetics ; Polymorphism, Genetic

OBJECTIVEHyperphenylalaninemia (HPA) is an inborn error of metabolism in which the hydroxylation of phenylalanine (Phe) to tyrosine is disturbed. Accumulation of Phe leads to severe mental and psychomotor retardation. (1)H magnetic resonance spectroscopy ((1)HMRS) is a novel non-invasive method to quantitate the brain metabolites besides Phe concentration in HPA patients. And it could be acquired conveniently on clinical MRI routine scanners. This study aimed to investigate the correlation between blood Phe ([Phe](blood)) and [Phe](brain), the characteristics of blood-brain Phe metabolism and its impacts on mental retardation.

METHODTotally 32 untreated patients diagnosed with HPA were studied, including 18 boys and 14 girls (age ranging from 33 days to 13 years). The patients were divided into two groups: elder than 4 months old (n = 22) and younger than 4 months old (n = 10). (1)HMRS were performed in all patients. [Phe](brain) were measured by absolute [Phe](brain) using Creatinine as an internal reference. [Phe](blood) were measured and developmental quotient (DQ) or intelligence quotients (IQ) were evaluated.

RESULT(1) [Phe](brain) measured by (1)HMRS ranged from 0.0640 to 0.6296 (M = 0.1542) while the [Phe](blood) was from 0.3804 to 2.5140 mmol/L (M = 1.5210 mmol/L) in all the 32 cases of HPA patients. (2) There was a positive linear correlation (r = 0.6103 (P < 0.01)) between [Phe](blood) and [Phe](brain). And there were interindividual differences in [Phe](brain) in several patients. (3) Variable mental retardation were observed in 23/32 cases in this study. (4) There was a negative correlation between [Phe](blood) and [Phe](brain) to the mental retardation (r(blood) = -0.5045, r(brain) = -0.6471 (P < 0.01)) in 22 cases of the HPA patients older than 4 months. And [Phe](brain) had more significant correlation with mental development than [Phe](blood).

CONCLUSIONThe [Phe](blood) could correspondingly represent the [Phe](brain) in most HPA patients. The Phe concentration could reflect the degree of mental retardation substantially in 22 cases with HPA older than 4 months. And the [Phe](brain) could more accurately illustrate it. (1)HMRS can be used to quantitate intracerebral Phe concentrations non-invasively in HPA patients. Preliminary findings suggest that interindividual variations in the kinetics of Phe uptake and metabolism do exist. (1)HMRS has great clinical significance in understanding the mechanism of HPA patient's mental retardation, providing proper objective standards for better diagnosis and treatment of HPA patients.

Adolescent ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Intelligence Tests ; Magnetic Resonance Spectroscopy ; Male ; Phenylalanine ; analysis ; Phenylketonurias ; metabolism