OBJECTIVE:To investigate the stability of injectable pantoprazole sodium in 4 kinds of infusions.METHODS: A HPLC method was adopted in this determination, the content change of injectable pantoprazole sodium was determined within 4 hours' mixing with 4 kinds of infusions and its change of color was observed; meanwhile the effects of pH value, potassium,magnesium,calcium ions on the stability of injectable pantoprazole sodium were investigated.RESULTS:No obvious changes were noted in terms of contents, appearances, pH value, UV - absorption maximum wavelength for the injectable pantoprazole sodium solution within 4 hours either in 4 kinds of common infusions or in potassium,magnesium,calcium ions water solutions.It was extremely unstable when pH value of pantoprazole sodium solution was lower than 7.0;when its pH value was 7.0, its color became yellowish yet without obvious loss of content; it is stable within 4h when its pH value was above 8.0.CONCLUSIONS:Injectable pantoprazole sodium remains stable within 4 hours either in 4 kinds of common infusions or in potassium, magnesium, calcium ions water solutions, the pH value has a great influence on the stability of pantoprazole sodium solution.

Objective To investigate the immunofluoresce nt and immunoblotting features of paraneoplastic pemphigus(PNP).Methods Sera were tested by indirect immunofluorecence(IIF)and immunoblotting(IB)in four patients with PNP.Results Binding of IgGand C3to the intercell ular spaces of epidermis was found in IIF with rat bladder as substrate.The titers decreased after the excision of tumo rs.The autoantibodies also bound to sub-strates of the epithelia of human ski n,rat tongue and esophagus in those p atients.The patients' sera recogni zed 210kD and 190kD antigens from human kera tinocyte extracts with IB analysis.Conclusions IIF with rat bladder as substrate can be used a screening test for PNP.PNP tends to involve epithelia of mucosa.The results of IB test m ay confirm the diagnosis of PNP of the 4p atients.[

Objective To investigate the protective effect and mechanism of pamidronate disodium on calcification in rat vascular smooth muscle cells (RVSMCs) induced by hyperphosphate.Methods RVSMCs were placed in various culture mediam, including normal phosphate medium (Pi 1.4 mmol/L), high phosphate medium (Pi 4.5 mmol/L), different pamidronate disodium concentrations medium (Pi 4.5 mmol/L+pamidronate disodium 10-5, 10-6, 10-7 mmol/L). Calcium content and cell protein content were quantified by the O-cresolphthalein complexone method and BCA protein assay respectively. Calcification was visualized by yon Kossa staining, and cbfα-1,osteocalcin were quantified by Western blotting. Results After culture for 3 days, calcium content in high phosphate group were much higher than that in control group, and pamidronate disodium groups had a lower calcium content compared with high phosphate group (all P＜0.05).Calcium deposit in RVSMCs was greater in high phosphate group, while pamidronate disodium groups revealed obviously decreased deposit (all P＜0.05). Protein expression of cbfα-1 and osteocalcin in pamidronate disodium groups was much lower than that in high phosphate group.Conclusion Pamidronate disodium can protect RVSMCs from phosphate-induced calcification in vitro, which may be associated with the blockage of transformation of RVSMCs into osteoblast.

Motivation ; Forensic Psychiatry ; Violence ; Wounds and Injuries ; Crime/psychology*

Objective To investigate the inhibitory effects of RNA interference(RNAi)on the expression of matrix metalloproteinase-9(MMP-9)gene and invasiveness and adhesion of ovarian cancer cells.Methods Four groups of different specific target sequence in coding region of MMP-9 and one non- specific sequence were chosen,which were Sitel,Site2,Site3,Site4 and Site5.Small interference RNA (siRNA)expression cassettes(SEC)were constructed by PCR and transfected into ovarian cancer HO- 8910PM cells.RT-PCR and western blot were used to detect mRNA and protein expression of MMP-9 gene; the abilities of invasion and adhesion were detected by Matrigel invasion assay and cell adhesion assay. Results The expression of MMP-9 was inhibited and the inhibitory effects of different sequence were varied.The mRNA expression was 0.64?0.06,0.47?0.07,0.55?0.10 in Sitel,Site2,Site3 group, and protein expression was 0.30?0.09,0.27?0.08,0.37?0.12,respectively.Site2 group had the most efficient inhibitory effect,followed by Sitel and Site3 groups.Cell growth curve revealed that cell growth was significantly inhibited in Site2 group.Invasiveness and adhesion were significantly reduced,the inhibitory rate on invasion in Site1,Site2,Site3 groups were 50.0%,50.0% and 37.5%,respectively;the inhibitory rate on adhesion in Site1,Site2,Site3,Site4 groups were 43.8%,48.8%,33.9%,24.2% at 60 min and 41.6%,40.2%,35.1%,16.0% at 90 min,respectively.Conclusions RNAi exists in ovarian HO-8910PM cells.MMP-9 siRNA can specifically down-regulate MMP-9 expression and lead to the inhibition of invasiveness and adhesion in ovarian cancer cells.

Objective To evaluate the long term effect of video-assisted thoracoscopic thymectomy for myasth,enia gravis and the influence of concomitant thymoma.Methods 47 cases of myasthenia gravis were retrospectively reviewed who had received video-assisted thoracoscopic thymectomy from Apr.2001 through Oct.2009.The patients were separated to two groups with or without thymoma.Influence of oncologic factors was carefully studied.Results There were 20 males and 27 females with a mean age of 36.6 yrs.According to the typing system of the Myasthenia Gravis Foundation of America ( MGFA),the patients belonged to type Ⅰ 18 cases,type Ⅱ a 14 cases,type Ⅱ b 14 cases,and Ⅲa 1 case.22 patients were in the group with thymoma,and the other 25 in the group without thymoma.Until the deadline of follow-up time of Jun.2011,only two cases in non-thymomatous group were lost.Follow-up time was 20 to 122 months,mean 57 months.The complete stable remission rate(CSR),pharmacologic remission(PR),minimal manifestations(MM),worse(W),exacerbation(E) and died of myasthenia gravis(D) in non-thymomatous group were 78.3％,13.0％,4.3％,0,0 and 4.3％.In thymomatous group the values were 50.0％,22.7％,13.6％,4.5％,9.1％ and 0.Conclusion Video-assisted thoarcoscopic thymectomy has a satisfactory long term effect for myasthenia gravis.Thymomatous group has no different in overall effectiveness with that of non-thymomatous group although a probably lower complete stable remission rate is prompted.

Objective To evaluate the degree of hypothermia cerebral resuscitation patients by acute physiology and chronic health evaluation Ⅱ (APACHE Ⅱ) score and judge the prognosis,so as to prove the effectiveness of its application.Methods Data of 34 cases of patients with mild hypothermia cerebral resuscitation in ECIU or ICU were continuously observed,calculated the scores of APACHE Ⅱ and verify for establishing the regression model.Results The APACHE Ⅱ score was 20-47 (33.86 ± 5.12) scores.The APACHE Ⅱ score in 9 patients who were survived within 72 h was (27.83 ± 4.89) scores,and in 25 patients who were dead within 72 h was (35.56 ± 7.12) scores,there was significant difference (P ＜ 0.01).Logistic regression analysis showed that APACHE Ⅱ score was the risk factor (P ＜ 0.01).Conclusion APACHE Ⅱ score can be applied to evaluate the degree of mild hypothermia cerebral resuscitation and prognosis,it can guide the clinical decision making.

Objective To investigate the effects of Saussurea involucrata extract pretreatment on the expression of the Toll-like receptor 4 (TLR4)/nuclear factor-κB (NF-κB) in focal cerebral ischemia/reperfusion in mice and its possible neuroprotective mechanism.Methods Seventy-two Kunming mice were randomly divided into four groups:sham operation,saline,Saussurea involucrata extract,and edaravone groups (n =18 in each group).Saussurea involucrata extract 0.8 g/kg was given intraperitoneally in the Saussurea involucrata extract group; edaravone 3 mg/kg was given in the edaravone group; and the same volume of saline was given in the saline group.A model of middle cerebral artery occlusion (MCAO) was induced after 7 days of continuous injection.Cerebral infarct volume was determined by 2,3,5-triphenyltetrazolium staining.Immunohistochemical staining was used to detect TLR4-positive cells in ischemic brain tissue.Reverse transcriptase polymerase chain reaction was used to detect the expression of TLR4/NF-κB mRNA.Results The cerebral infarct volume in mice in the saline,Saussurea involucrata extract and edaravone groups was 131.55± 28.25 mm3,84.10 ±13.92 mm3 and 65.10 ± 6.78 mm3,respectively.There were significant difference (F =10.158,P =0.012).The infarct volume in the Saussurea involucrata extract group (P =0.020) and edaravone group (P0.005) was significantly less than that in the saline group,and there was no significantly difference between the 2 groups.The numbers of cortex and TLR4 positive cells in hippocampus area at the ischemic sides in the saline group were significantly more than those in the sham operation group (all P ＜0.001).The numbers of positive cells of cortex and TLR4 in the Saussurea involucrata extract group and the edaravone group were significantly decreased compared to the saline group (all P ＜ 0.05),and there was no significant differences between the Saussurea involucrata extract group and the edaravone group.The expressions of TLR4,p50,and p65 mRNA in the saline group were significantly up-regulated compared to the sham operation group (all P =0.000).Saussurea involucrata extract could significantly down-regulate the expressions of TLR4,p50,and p65 mRNA at 24 hours after ischemia/reperfusion (all P =0.000).Edaravone could significantly down-regulate the expressions of TLR4 and p65 mRNA (all P =0.000) and it had a down-regulated trend for the expression of p50 mRNA (P =0.053); while there was no significant difference in the expressions of TLR4 and p65 mRNA between the Saussurea involucrata extract group and the edaravone group.Conclusions Saussurea involucrata extract pretreatment may significantly reduce the cerebral infarct volume,down-regulate the expressions of TLR4 and NF-κB subunit,and play a neuroprotective effect by inhibiting inflammatory response after ischemia.

OBJECTIVETo observe the inhibitory effects of antisense MMP-9 oligodeoxynucleotides on invasiveness and adhesion ability in vitro of ovarian cancer cells, and to investigate the mechanisms of action.

METHODSMMP-9 antisense oligonucleotides were transfected by lipofectinmin into ovarian cancer cell line HO-8910PM cells expressing MMP-9 induced with fibronectin. RT-PCR, Western blot and gelatin zymography were used to detected MMP-9 expression of mRNA and protein and enzymatic activity. The ability of invasion and migration of ovarian cancer cells was assayed in Transwell cell culture chamber. Cell adhersion assay was carried out in a microculture well pre-coated with fibronectin.

RESULTSMMP-9 expressions of mRNA and protein were significantly decreased in the antisense-transfected cells. Comparing with the control group, the inhibition rate was 34. 8% and 42. 5% , respectively (P <0. 05). Its gelatin enzymatic activity was inhibited. Matrigel invasion assay and Transwell migration assay revealed markedly reduction in invasion and migration for the antisense group. The inhibition rates were 22. 4% and 24. 8% , respectively. The adhesion ability was also reduced. The inhibition rates were 49. 8% and 38. 3% at 60 min and 90 min, respectively.

CONCLUSIONMMP-9 down-regulation can significantly inhibit the ability of invasion and attachment of ovarian cells in vitro. MMP-9 may play an important role in invasion and metastasis of ovarian cells and potentially be a molecular target of blocking invasion and metastasis of ovarian cancer.

Blotting, Western ; Cell Adhesion ; genetics ; physiology ; Cell Line, Tumor ; Cell Movement ; genetics ; physiology ; Down-Regulation ; Female ; Humans ; Matrix Metalloproteinase 9 ; genetics ; metabolism ; Neoplasm Invasiveness ; Oligodeoxyribonucleotides, Antisense ; genetics ; Ovarian Neoplasms ; enzymology ; genetics ; pathology ; RNA, Messenger ; genetics ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction ; Transfection

BACKGROUNDEndometrial carcinoma is one of the most common female tract genital malignant tumors. Nifedipine, an L-type calcium channel antagonist can inhibit cell proliferation of carcinomas. Recent studies indicated that a rise in the free cytosolic calcium ([Ca(2+)](c)) was a potent inducer of autophagy. Here, we investigated the relationship between nifedipine and autophagy in Hec-1A cells.

METHODSCells were cultured with nifedipine (10 µmol/L) and harvested at different times for counting cell number. MTT assay was applied to evaluate the cell viability and transwell assay to reveal cell migration. Apoptotic cells were detected with annexin V/PI assay. Then cells were treated with 3-methyladenine (3-MA) (2.5 mmol/L) for 0, 5, 15, 30, 60, and 120 minutes and the expression of the L-type calcium channel alpha1D (Cav1.3) protein was detected. At last, cells were cultured and assigned to four groups with different treatment: untreated (control group), 10 µmol/L nifedipine (N group), 2.5 mmol/L 3-MA (3-MA group), and 10 µmol/L nifedipine plus 2.5 mmol/L 3-MA (N+3MA group). Autophagy was detected with GFP-LC3 modulation by fluorescent microscopy, and expression of the autophagy-associated proteins (LC3, Beclin1 and P70s6K) by Western blotting and monodansylcadaverine (MDC) labeled visualization.

RESULTSProliferation of Hec-1A cells was obviously suppressed by nifedipine compared with that of the untreated cells for 24, 48, and 96 hours (P = 0.000 for each day). The suppression of migration ability of the nifedipine-treated cells (94.0 ± 8.2) was significantly different from that of the untreated cells (160.00 ± 9.50, P = 0.021). The level of early period cell apoptosis induced by nifedipine was (2.21 ± 0.19)%, which was (2.90 ± 0.13)% in control group (P = 0.052), whereas the late period apoptosis level reached (10.38 ± 0.96)% and (4.40 ± 0.60)% (P = 0.020), respectively. The 3-MA group induced a slight increase in the Cav1.3 levels within 15 minutes, but significantly attenuated the Cav1.3 levels after 30 minutes. There were more autophagic vacuoles labeled by MDC in the N group (20.63 ± 3.36) than the control group (6.29 ± 0.16, P = 0.015). GFP-LC3 localization revealed that the LC3 levels of cells in 3-MA group, N+3MA group, 3-MA group were 2.80 ± 0.29, 2.30 ± 0.17, and 1.80 ± 0.21, respectively. Cells in the N group showed significant augmentation of autophagy (P < 0.05). Western blotting analysis confirmed the down-regulation of LC3 levels in 3-MA group (0.85 ± 0.21) and N+3MA group (1.21 ± 0.12) compared with nifedipine treatment (2.64 ± 0.15, P < 0.05). The annexin-V-FITC/PI assay showed that the level of early period cell apoptosis induced in the N+3-MA group ((11.22 ± 0.91)%) differed significantly from that of the control group ((2.51 ± 0.70)%) and N group ((3.47 ± 0.39)%). Similarly, the late period level of the N+3-MA group ((55.19 ± 2.51)%) differed significantly from that of the control group ((15.81 ± 1.36)%) and the N group ((22.09 ± 2.48)%, P < 0.05). The down-regulated expression of P70s6k and up-regulated expression of the Beclin1 revealed significant differences between the N+3-MA group and control group (P = 0.025; Beclin1: P = 0.015).

CONCLUSIONSProliferation and migration in vitro of endometrial carcinoma Hec-1A cells are significantly suppressed by nifedipine. The nifedipine leads autophagy to oppose Hec-1A cells apoptosis. Autophagy inhibition by 3-MA leads down-regulation of Cav1.3 and enhances nifedipine-induced cell death. The nifedipine-induced autophagy is linked to Beclin1 and mTOR pathways.

Adenine ; analogs & derivatives ; pharmacology ; Apoptosis Regulatory Proteins ; physiology ; Autophagy ; drug effects ; Beclin-1 ; Calcium ; metabolism ; Calcium Channel Blockers ; pharmacology ; Calcium Channels, L-Type ; physiology ; Cell Line, Tumor ; Endometrial Neoplasms ; drug therapy ; pathology ; Female ; Humans ; Membrane Proteins ; physiology ; Nifedipine ; pharmacology ; Signal Transduction ; physiology ; TOR Serine-Threonine Kinases ; physiology