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Objective To study the expression and biological functions of RN181 in SMMC7721 cells,the retroviral vector was constructed.Methods Gene cloning techniques were used to construct pRetrox-TRE3G/RN1S1 recombinant vector.The regulating plasmid pRetroX-TRE3G/RN181 or the response plasmid of pRetroX-Tet3G were respectively cotansfected into GP2-293 cells with Envelope Vector plasmid to package retrovirus after routine identification.Both viruses co-infected target cells SMCC7721,and then were selected by G418 to obtain stable cell lines.The stable cell lines were induced by doxycycline (DOX),and then verified by RT-PCR and Western blotting.CCK-8 was used to evaluate the effect of RN181 on growth of SMMC7721 cells.Results We constructed the recombinant plasmid.Stable recombinant plasmid were verified by screening.And there were significant differences of RN181 between the induced and uninduced cell lines through RT-PCR and Western blot.Conclusions We have successfully constructed the inducible stable RN181 expression SMCC7721 cell,which can be used as an effective cell model to study the biological functions of RN181.We found RN181 could suppress the proliferation and invasion in SMMC7721 cells in vitro.
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Purpose To investigate the diagnostic criteria and value of angle parameters in female stress urinary incontinence (SUI),and to provide an objective basis for clinical diagnosis of SUI.Materials and Methods In retrospective analysis,138 patients with SUI diagnosed by clinic from December 2014 to December 2016 were selected as the study group,while 100 normal females receiving the routine medical examination were selected as the control group.All subjects in both groups were examined by pelvic floor ultrasound to observe the urethral inclination angle,urethral rotation angle (URA) and posterior urethrovesical angle (PUVA) to obtain the standard in diagnosis of female SUI.30 SUI patients from January to February 2017 were prospectively analyzed,and the above results were verified to evaluate their value in the diagnosis of SUI.Results There was no significant difference in age,body mass index,gravidity and parity history and resting state PUVA between the two groups (P＞0.05).There was significant difference in the resting state urethral inclination angle,Valsalva urethral inclination angle,PUVA and URA between the two groups (P＜0.05).The receiver operating characteristic curve was used to find the cut-off value of SUI:urethral inclination angle in resting state ＜14.5°,urethral inclination angle in Valsalva state ＞32.5 °,and URA ＞49.5 °,PUVA ＞154.5 °,with their sensitivity of 62％,73％,67％ and 57％,respectively;the specificity of 61％,84％,82％ and 70％,respectively;and the area under the curve of 0.633,0.809,0.768 and 0.658,respectively.By verification,when the urethral inclination angle was 14.5° in the resting state,the coincidence rate of the diagnosis of SUI was 63％;when the urethral inclination angle of Valsalva was 32.5°,83％;when the URA was 49.5°,77％;when the PUVA was 154.5 °,60％.Conclusion Pelvic floor ultrasound can clearly observe angle parameters of females,the determination of the cut-off value of ultrasound is helpful for clinical diagnosis and evaluation of SUI,and the urethral inclination angle in Valsalva state and URA are of high diagnostic value in the diagnosis of female SUI,providing an objective basis for clinical diagnosis of SUI.
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Objective To study the clinical results of patients with gallbladder stones and common bile duct stones treated either by laparoscopic cholecystectomy followed by endoscopic sphincterotomy (EST + LC) or laparoscopic cholecystectomy + common bile duct exploration (LC + LCBDE) in the elderly patients.Methods A retrospective study was conducted on 96 patients who had common bile duct and gallbladder stones treated from January 2012 to January 2016.The patients were divided into the control group and the observation group.46 patients were in the observation group who underwent LC three days after EST,while the remaining patients were in the control group who underwent LC and LCBDE.The serum amylase levels before LC,operation time,intraoperation bleeding volume,postoperative time to first flatus,total hospitalization stay and total hospitalization costs and incidences of postoperative complication were compared.Results The total operation time was (95.0 ±7.0) minutes and (125.0 ± 18.0) minutes,respectively,(P＜0.05).The total costs in the two group were (39515.0 ±4 135.0) yuan and (28287.0 ± 2 254.0) yuan (P ＜ 0.05),respectively.Postoperative complications were observed in 5 (6.1％) and 10 (13.2％) patients (P ＜ 0.05),respectively.The preoperative serum amylase levels were (97.6 ± 48.5) IU/L and (131.4 ± 68.7) IU/L,respectively.The blood loss was (35.7 ± 8.5) ml and (31.8 ± 7.3) ml,respectively.The postoperative time to first flatus was (1.7 ± 0.5) days and (1.9 ± 0.4) days,respectively.The total hospitalization stay was (16.3 ±2.8) days and (15.2 ±3.7) days.There were no significantly differences (P ＞ 0.05).Conclusions LC carried out on day 3 after EST to treat elderly patients with cholecystolithiasis and choledocholithiasis was safe and efficacious and the treatment had the advantages of minimal trauma,short operative time,rapid recovery and low complication rates.This should be recommended in clinical practice.
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Objective To determine whether it is better to carry out laparoscopic cholecystectomy (LC) immediately or 3 days after endoscopic sphincterotomy (EST) in patients with common bile duct stones and gallbladder stones.Methods A retrospective study was conducted on 160 patients who had common bile duct stones and gallbladder stones treated from July 2013 to July 2015 in the First People's Hospital of Huizhou.These patients were divided into a control group and an observation group,with 80 patients in each group.The control group underwent LC immediately after EST,while the observation group underwent LC 3 days after EST.The levels of blood amylase before LC,operation time,intraoperative bleeding volume,postoperative time to first flatus,postoperative complication after EST,postoperative hospitalization stay,total hospitalization stay and total hospital costs and incidences of postoperative complication after LC were compared between the two groups.Results The time periods of total hospitalization stay in the observation group and the control group were (18.2 ±3.5) d and (12.3 ±3.0) d,respectively (P＜0.05).The total costs in the two groups were (32 164 ±9 125) RMB and (22 375 ±7 860) RMB,respectively (P ＜ 0.05).The overall incidences of postoperative complication [patients (％)] were 5 (6.3) and 9 (11.3),respectively (P ＜ 0.05).The differences on preoperative serum amylase levels,LC operative time,blood loss,postoperative time to first flatus and postoperative hospitalization stay were not significantly different (P ＞ 0.05).Conclusion LC carried out on day 3 after EST to treat common bile duct stones and gallbladder stones was clinically more efficacious.
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Objective To evaluate the preoperative liver function and prognosis of laparoscopic cholecystectomy (LC) in patients with cirrhosis,using the Child-Turcotte-Pugh classification and the model for end-stage liver disease(MELD) score.Methods From January 2009 to June 2013,973 patients who were admitted to the Department of General Surgery of our hospital and the HuiZhou Municipal Central Hosptial were studied.Of the 373 patients with cirrhosis,38 patients were excluded because of Child C,MELD ＞ 30,or laparotomy.The remaining 335 patients who received laparoscopic cholecystectomy were randomly divided into two groups The Child grade and MELD score were retrospectively analyzed.Results There was no significant difference in intraoperative hemorrhage between the Child A group [(106 ± 11) ml] and the Child B group [(109 ± 11) ml] (P ＞ 0.05).The R ＜ 14 scores in the MELD group [(58 ± 15) ml] was significantly lower than that in the R≥ 14 group [(120 ± 28) ml] (P ＜ 0.01).There was no significant difference in postoperative complications between the Child group A (10 cases,12％) and the Child group B (17 cases,21％) (P ＞0.05).There was a significantly lower incidence in the R ＜ 14 scores in the MELD group (10 cases,12％) than the R ≥ 14 group (27 cases,33％) (P ＜ 0.05).There was also no significant difference in the hospital stay between the Child A group (9 ± 1) and the Child B group (10 ± 2)(P ＞0.05) ; the R ＜ 14 score of the MELD group (7 ± 1) was significantly less than that of the R≥ 14 group (11 ±2) (P ＜0.01).There was no significant difference in the cost of hospitalization between the Child A group (1.337 ± 0.063) and the Child B group (1.359 ± 0.089) (P ＞ 0.05) ; the R ＜ 14 group (MELD score 1.108 ± 0.123) was significantly less than that of the R ≥ 14 group (1.568-± 0.117)(P ＜ 0.01).Conclusion Compared with the Child-Turcotte-Pugh classification,the MELD score was more scientific,objective and accurate in judging the preoperative liver function.It helped to predict the amount of intraoperative hemorrhage and postoperative morbidity,reduced hospital stay and hospitalization expenses.Therefore,the MELD scoring system more objectively guided the treatment of patients with cholecystitis with cirrhosis.
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Objective Within human hepatoma cell lines,we aimed to investigate the effects of the down-regulation by RNAi on different fragments of osteopontin (OPN) in order to discover more effective and accurate sites for OPN.Methods Specific small interfering RNA of OPN (OPNi-1) were synthesized and transfected into human hepatoma cell line (HEP-G2).Fluorescent quantitative PCR and immunohistochemical methods were used to test*the OPN expression levels of mRNA and protein before and after RNAi.Results After transfection,the △CT value of the A fragment was greater than B and C fragments of OPN mRNA in HEP-G2.Before RNAi was added to HEP-G2 cells,the three fragments A,B,C had OPN mRNA CT values of 8.31±1.58,8.78±1.49,8.25±1.51 respectively.Once the RNAi were added,the CT values were measured 48h after for the fragments A,B,and C which were 12.14±1.43,10.22±1.97,10.48±1.88 (P＜0.05) respectively.The immunohis tochemical values of A,B,C were down from 6.44±1.67,5.43±2.05,5.45±2.52 to 2.84±1.52,4.43± 1.65,3.95± 1.43 respectively after interference.Conclusions RNAi can inhibit the expression of OPN gene selectively.siRNA targets different segments of OPN,which may have more effects on invasion and metastasis of liver cancer for a more important significance in science and health economics.
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<p><b>OBJECTIVE</b>To investigate the fragmentation pathway of vitexin and isorhamnetin-3-O-beta-D-rutinoside with CID-TOF-MS.</p><p><b>METHOD</b>Equipped with an LC-MS was carried out using an ultra-performance liquid chromatography, electrospray ionization quadrupole collision-induced dissociation-TOF-MS.</p><p><b>RESULT</b>ESI-MS spectrum showed [M-H]- base peak of m/z 431. 0958 and m/z 623.1566. The CID-MS of vitexin showed five basic fragment ions, three of which corresponded to the glucosyl ring fracture: m/z 353, 341 and 311; other two were benzyl ion m/z 283, aglycone ion m/z 269. In addition, two low abundance ions, namely, m/z 161 and m/z 117, generated by RDA cracking ions, were also characteristic ions. The CID-MS of isorhamnetin-3-O-beta-D-rutinoside showed six main characteristic fragments ions corresponding to the loss of rhamnosyl m/z 477 and the glycosyl ring fracture: m/z 387, 357 and 311, and aglycone ion m/z 315. In addition, B ring generated m/z 300 and m/z 271 and C ring generated m/z 243 and the RDA cleavage generated m/z 151 and m/z 125.</p><p><b>CONCLUSION</b>Those fragment ions can be used to quickly identify vitexin and isorhamnetin-3-O-beta-D-rutinoside.</p>
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Apigenin , Chemistry , Disaccharides , Chemistry , Drugs, Chinese Herbal , Chemistry , Flavonoids , Chemistry , Molecular Structure , Spectrometry, Mass, Electrospray Ionization , MethodsABSTRACT
<p><b>OBJECTIVE</b>To establish the mathematical kinetic model of the components extracted from the Salvia miltiorrhiza.</p><p><b>METHOD</b>In the conditions of ultrasound extracting the course of traditional Chinese medicine, the role of ultrasound intensity is large enough, the proliferation of boundary layer can be infinitesimal, as well as the solute upon the surface of medicine quickly into the main solution, we can assume that the whole process of extraction from the control of proliferation. A mathematical model of dual-frequency ultrasound extraction kinetics based on Fick's second diffusion law was established for traditional Chinese medicine. Based on material size and solid/liquid ratio significant factors.</p><p><b>RESULT</b>We adopt Origin software to carry out a mathematics simulation, the result showed: simulation graphics and experiment value are very fitting.</p><p><b>CONCLUSION</b>This proves that S. miltiorrhiza extraction course accords with ordinary extraction dynamics equation at the dual-frequency ultrasound.</p>
Subject(s)
Drugs, Chinese Herbal , Chemistry , Kinetics , Models, Theoretical , Particle Size , Plants, Medicinal , Chemistry , Salvia miltiorrhiza , Chemistry , UltrasonicsABSTRACT
Objective To establish a HPLC-ESI-MS method for simultaneous analysis and determination of flavonoids in Semen Hoveniae. Methods The separation was performed on Acquity UPLCTM BEH C18 column (50 mm× 2. 1 mm,1.7 μm) with a mobile phase of formic acid aqueous solution (B) and MeOH (A). The flow rate was at 1.0 mL/min and detection wavelength was at 290 nm. Samples were analyzed in positive-ion detection model. Results Five kinds of flavonoids in Semen Hoveniae were separated well and were tentatively identified by ESI-MS and by reference to litera-ture data. Conclusion The method is accurate and rapid, and can be used for the identification of flavonoids in Semen Hoveniae.
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AIM: To set up a rapid analysis and characterization method for coumarins in extract of Cnidium monnieri. METHODS: The sample was analyzed by ultra-performance liquid chromatography with a reversed phase C_(18) column using a binary eluent of acetonitrile and water(with 0.1% formic acid) under gradient conditions.The ~+ ions of coumarins in Cnidium monnieri extract were observed by positive ion electrospray time of flight mass spectrometry (ESI-TOF-MS) online detection,and the fragmentation behavior obtained by ESI-TOF-MS/MS,then coumarins was identified through accurate molecular weight and molecular formula and fragmentation information combined with literature review. RESULTS: All components could be separated rapidly within 10 min,and five coumarins-xanthotoxol,bergapten,xanthotoxin,imperatorin and osthol could be primary identified in Cnidium monnieri extract. CONCLUSION: This method is effective for identification of coumarins in extract of Cnidium monnieri.