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

2,4-Dichlorophenoxyacetic Acid ; analogs & derivatives ; toxicity ; Animals ; Epididymis ; growth & development ; metabolism ; Female ; HSP70 Heat-Shock Proteins ; biosynthesis ; genetics ; Herbicides ; toxicity ; Male ; Rats ; Rats, Wistar ; Spermatozoa ; drug effects ; growth & development ; metabolism

Flavonol synthase (FLS) is one of the key enzymes in flavonoids metabolic pathways. In this study, middle sequence was obtained from Carthamus tinctorius transcriptome sequencing results. Full-length cDNAs of FLS was cloned from petals of C. tinctorius to FLS by using RT-PCR and RACE technology. Its full-length cDNA was 1,201 bp, with an open reading frame of 1,101 bp and 336 encoded amino acids. The phylogenetic analysis showed that, FLS gene encoded amino acids in C. tinctorius were highly homologous with amino acids in congeneric Compositae species, especially Rudbeckia laciniata. The pBASTA-FLS plant expression vector was successfully built by the molecular biology method, which lays a foundation for further studying biology functions of the gene and biosynthesis mechanism of flavonoids.
Amino Acid Sequence ; Base Sequence ; Carthamus tinctorius ; classification ; enzymology ; genetics ; Cloning, Molecular ; DNA, Complementary ; genetics ; metabolism ; Molecular Sequence Data ; Open Reading Frames ; Oxidoreductases ; genetics ; metabolism ; Phylogeny ; Plant Proteins ; genetics ; metabolism

To investigate the influence of Anxin granules combined with tirofiban on acute myocardial infarction (AMI) Patients after elective percutaneous coronary intervention (PCI). One hundred and twenty AMI patients were randomly divided into treatment group and control group. The patients in the two groups were all given Tirofiban 30mins before PCI . The treatment group was added Anxin granules 30 mins before and after PCI. Tissue factor (TF) and von willebrand factor (vWF) were tested at 6 hours after operation. Syndromatology alteration of traditional Chinese medicine (TCM) and bleeding complications were observed at 4 weeks after operation. Both TF and vWF at 6 hours after operation of the treatment group was lower than the control group significantly (P < 0.01), while the condition of myocardial ischemia at 90 mins after operation of the treatment group was better than control group with significance. The syndromatology alteration of TCM especially spontaneous perspiration and hypodynamia of the treatment group were improved significantly compared to control group 4 weeks after operation. All patients in both groups had no bleeding complications and thrombopenia. The study suggests that Anxin granules combined with tirofiba can improve the clinical efficacy and the endothelial function of AMI patients after PCI with no increase in bleeding events.
Aged ; Angioplasty, Balloon, Coronary ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Humans ; Middle Aged ; Myocardial Infarction ; complications ; metabolism ; surgery ; Postoperative Hemorrhage ; drug therapy ; etiology ; metabolism ; prevention & control ; Thromboplastin ; metabolism ; von Willebrand Factor ; metabolism