1.Oxidized related substance of methionine in compound amino acid injection (18AA-Ⅱ)
Tianshu SHAO ; Changming ZHOU ; Hui LI ; Lei GUO
Journal of China Pharmaceutical University 2021;52(6):713-718
TA method for the content determination of methionine sulfoxide and methionine sulfone in compound amino acid injection (18AA-II) was established in order to investigate their level in 155 batches of this product, and to explore the reason for the generation of these two impurities.The determination was performed on an Agilent Poroshell 120 EC-C18 column with mobile phases of sodium acetate/tetrahydrofuran solution (A) and sodium acetate solution -acetonitrile-methanol (B, 200∶400∶400) (gradient elution) at the flow rate of 0.5 mL/min.The excitation wavelength and the emission wavelength of the fluorescence detector were 233 nm and 441 nm, respectively.The column temperature was 40 °C, and the injection volume was 8 μL.The contents of methionine sulfoxide and methionine sulfone from 155 batches of compound amino acid injection (18AA-II) was determined using this method, and the residual oxygen content was detected by headspace gas analyzer.The results showed that the linear range of methionine sulfoxide and methionine sulfone were 0.128 1-10.250 0 μg/mL (r = 0.999 9) and 0.261 0-10.440 0 μg/mL (r = 0.999 8), respectively.The limits of quantitation were 0.13 μg/mL and 0.26 μg/mL, respectively; the limits of detection were 0.04 μg/mL and 0.09 μg/mL, respectively.RSDs of precision, stability and repetitive test were all lower than 1.3%.The recoveries ranged 98.00%-100.79% (RSD = 1.15%, n = 9) and 98.19%-102.31% (RSD = 1.33%, n = 9).The content level of oxidized related substances from different manufacturers showed significant difference, showing relevance with the residual oxygen content to some extent, yet no significant correlation with the added amount of antioxygen (sodium pyrosulfite).The method is validated to be useful for the content control of methionine sulfoxide and methionine sulfone in compound amino acid injection (18AA-II).It is quite necessary to include the determination of oxidized related substance into the quality specification.Manufacturers should strengthen the control of remaining oxygen in their products.
2.Clinical Effects of Levofloxacin Eyedrops in the Treatment of Acute Bacterial Conjunctivitis and Keratitis
Changming YUAN ; Jiawen LI ; Liling ZHANG ; Qiaoshan GUO ; Yan YANG
China Pharmacy 2001;12(6):264-265
OBJECTIVE: To observe the therapeutic effects of 0.3% levofloxacin eyedrops on the acute bacterial conjunctivitis and keratitis.METHODS: The patients were randomly divided into treated group and control group.They were given 0.3% levofloxacin and 0.3% ofloxacin eyedrops respectively.The effects were compared between two groups.RESULTS: From April 1998 to December 1998, 132 cases were enrolled in the study: treated group 67 cases, control group 65 cases.The positive rates of bacterial culture were over 80% in both groups.It showed that there was no significant difference between two eyedrops in the curative and effective rates, but the mean cure day of keratitis in levofloxacin group was significantly shorter than that in ofloxacin group.CONCLUSION: Levofloxacin eyedrops is effective and safe in treating bacterial conjunctivitis and keratitis, and the therapeutic course is short and no obvious ARDs occur.
3.Effect of gynecological laparoscopic surgery in different tidal volume mechanical ventilation on lung injury
Lingli WANG ; Changming YANG ; Longquan XIANG ; Wei GE ; Xiaoli GUO
Chinese Journal of Primary Medicine and Pharmacy 2017;24(19):2941-2945
Objective To investigate the effects of different tidal volume mechanical ventilation on pulmonary function and inflammatory factors in gynecologic laparoscopic surgery.Methods 45 patients undergoing gynecological laparoscopic surgery without lung disease,ASA Ⅰ-Ⅱ,aged 20-45 years old,weight 45-65 kg,operation time 2-3 hours,were randomly divided into A,B,C three groups,15 cases in each group.Mechanical ventilation with different tidal volume of the three groups,A group,B group and C group were 8mL/kg,9mL/kg,10mL/kg.Blood gas analysis was performed before anesthesia (T0),30 minutes (T1),(T2),PA-aDO2 (60 minutes) (T3),and 120 min (T4),respectively,and the peak airway pressure and mean pressure were recorded.IL-6,TNF-α and IL-10 levels in plasma by radial artery blood sampling were simnultaneously determined.Results Comnpared with A group,the concentrations of Ppeak and Pmean were increased in B group at T1-T4 (t =5.13,4.78,6.54,5.32 and t =7.54,4.88,5.37,4.95;all P < 0.05),the concentration of Ppeak and Pmean in C group at T1-T4 increased (t =7.76,8.87,7.23,8.99 and t =6.42,7.38,7.62,9.86;all P < 0.05).Compared with B group,the concentrations of Ppeak and Pmean were increased in C group at T1-T4 (t =4.76,5.87,4.23,3.99 and t =4.76,3.99,6.06,4.52;all P < 0.05).Compared with A group,the A-aDO2 values of B group and C group were increased at T1,T2,T3 and T4 (t =5.32,5.48,4.88,5.69 and t =7.85,7.32,8.45,6.67;all P < 0.05).Compared with B group at the same time point,the A-aDO2 value of C group increased at four times of T1-T4 (t =5.62,4.38,6.94,4.22,P <0.05).Compared with group A,the concentration of A-aDO2 in C group was higher than that in B group at the same time point (t =4.45,4.87,5.32,4.79 and t =7.68,7.59,7.44,8.38;all P < 0.05).The levels of IL-6 and TNF-α in C group were significantly higher than those in the control group (t =4.78,5.56,7.62,8.03 and t =3.98,4.52,5.46,6.23;all P < 0.05).Compared with T0,IL-6 and TNF-α concentrations AT T2-T4 in A group had no statistically significant differences (all P > 0.05).Compared with B group,the levels of IL-6 and TNF-α in C group were significantly higher (t =4.58,4.99,6.53,4.77 and t =5.62,7.89,6.43,4.52;all P < 0.05).Compared with T0,IL-6 and TNF-α concentrations at T2-T4 in A group had no statistically significant differences (all P >0.05).There was no significant difference in IL-10 concentration among the thrce groups.Conclusion Laparoscopic surgery according to the end tidal carbon dioxide partial pressure (PetCO2) ventilation frequency,8 mL/kg tidal volume mechanical ventilation has no effect on the IL-6,TNF-alpha,IL-10 and other inflammatory factors,mechanical ventilation tidal volume is more appropriate.
4.Effects of long-term exposure to high altitudes on perceptual closure
Fumei GUO ; Changming WANG ; Getong TAO ; Xinjuan ZHANG ; Hailin MA ; Yan WANG
Chinese Journal of Behavioral Medicine and Brain Science 2021;30(5):446-451
Objective:To investigate the neural mechanism of long-term exposure to high altitudes environment on the processing stage of perceptual closure.Methods:A cross-sectional study was conducted. Thirty college students who first entered the plateau area (at an altitude of 3 658 m) and had lived in high-altitude areas for two years were selected as high altitude group (HA group), and 29 college students matched in age, gender and education level who had never been to the plateau area were selected as sea level group (SL group). The differences of basic physiological parameters were compared between the two groups.The face pairwise comparison paradigm was applied in the subjects of the two groups and the differences between the two groups were compared by event-related potentials (ERPs) technology. SPSS 22.0 software was used for statistical analysis, and the mean ± standard deviation was used for statistical description. Independent sample t test was used for comparison of physiological data between the two groups, and repeated measurement variance analysis was used for ERP data. Results:The physiological results revealed that the HA group had a higher pulse rate((86.71±10.82)/min, (75.97±11.28)/min ; t=-3.19, P=0.002) and diastolic blood pressure than the SL group ((75.93±9.19)mmHg, ( 68.59±11.42) mmHg ; t=-3.20, P=0.002). The oxygen saturation level was significantly lower than that of SL group ((90.77±2.25)%, (98.31±1.56) %) ; t=14.00, P<0.001). In the face pairwise comparison paradigm test, the main effect of P1 latency of ERP was significant.Compared with SL group, HA group showed significantly shortened latency of P1 ((105.10±15.59) ms, (128.35±14.40)ms, P<0.001). The main effect of group of N170 amplitude was significant, HA group was larger than that of SL group((-7.57±3.83) μV, ( -5.11±3.26) μV, P=0.005). The interaction effect between group and hemisphere of NCL amplitude was significant ( F(1, 57)=9.72, P=0.003). Simple effect test showed that the amplitude on the right hemisphere of SL group was significantly larger than that on the left hemisphere((0.46±1.31) μV), (1.16±1.33) μV ). The latency of P1 was significantly correlated with heart rate( r=-0.46, P<0.01), oxygen saturation level( r=0.64, P<0.01) and diastolic blood pressure( r=-0.26, P=0.049). Conclusion:After long-term exposure to high altitude, the neural response related to early visual perception is faster. More psychological resources are recruited during the face structural encoding stage. Contra-lateral compensating effect appeares in the stage of perceptual closure.
5.Effects of propofol on phosphorylation of AMPA receptor GluR1 subunits in a rat model of visceral pain
Changming YANG ; Ruijuan GUO ; Yun WANG ; Rong SHI ; Anshi WU ; Yun YUE
Chinese Journal of Anesthesiology 2012;32(5):563-565
Objective To investigate the effects of propofol on the phosphoryhtion of α-amino-3-hydroxy5-methyl-4-isoxazolepropionate(AMPA)receptor GluR1 subunits at Serine-831 and Serine-845 sites in the spinal cord dorsal horn in a rat model of visceral pain.Methods Thirty male SD rars,aged 6-8 weeks,weighing 200300 g,in which intrathecal catheters were successfully placed without complications,were randomly divided into 3 groups(n-=10 each):sham operation group(group Ⅰ),visceral pain group(group Ⅱ)and propofol group (group Ⅲ).Visceral pain was induced by injection of 10% capsaicin 50μl via the rectum in groups Ⅱ and Ⅲ.While the equal volume of normal saline was given instead in group Ⅰ.Group Ⅲ received intrathecal injection of propofol 20 tg at 10 min before injection of capsaicin.While the equal volume of dimethyl sulfoxide was given instead of propofol in groups Ⅰ and Ⅱ.The cumulative pain score was recorded during 30 min after capsaicin injection.The rats were then sacrificed,and the lumbar segment(L3-6)of the spinal cord was removed for determination of the expression of GluR1 subunits and phosphorylation of GluR1 subunits at Serine-831 and Serine-845 sites in the spinal cord dorsal horn.Results Compared with group Ⅰ,the cumulative pain score and phosphorylation of GluR1 subunits at Serine-831 sites and Serine-845 in the spinal cord dorsal horn were significantly increased(P <0.05 or 0.01),but there were no significant differences in the expression of GluR1 subunits in groups Ⅱ and Ⅲ (P > 0.05).Compared with group Ⅱ,the cumulative pain score and phosphorylation of GluRl subunits at Serine831 and Serine-845 sites in the spinal cord dorsal horn were significantly decreased in group Ⅲ(P < 0.05 or 0.01).Conclusion Propofol can attenuate the visceral pain through the inhibition of the phosphorylation of AMPA receptor GluR1 subunits at Serine-831 and Serine-845 sites in tte rat spinal cord dorsal horn.
6.The study of relevance between platelet activity and HS1 phosphorylation in sepsis
Lei XU ; Dongfeng GUO ; Guorong LIU ; Qin SHI ; Changming ZHAO ; Min HANG
Chinese Journal of Emergency Medicine 2015;24(11):1253-1256
Objective To explore the change of function and expression of hematopoietic lineage cell specific protein-1 (HS1) and phosphorylated HS1 (p-HIS) and factors devoting to HS1 phosphorylation in platelet with sepsis.Methods Plasma with rich platelet was collected from 150 sepsis patients and 50 healthy subjects, and comparison of platelets adhesion and aggregation were detected by micro-pore method and platelet aggregation instrument.Meanwhile the ATP concentrations of washed platelet of two groups were detected by the kit to compare release reaction.And then total HS1 (t-HIS) and p-HS1 of platelet from two groups were compared by using western blot.Afterwards the specific inhibitors of Src and Syk were used to verify the HS1 activation regulated by Src and Syk in LPS-induced cell model.Results The significant differences were present between healthy subjects and sepsis patients in platelet counts, platelet distribution width (PDW) and mean platelet volume (MPV) (P < 0.01).The data showed the sepsis patients had greater ability than healthy subjects in adhesion, aggregation and release reaction.Meanwhile the platelets of sepsis patients had higher concentration of t-HS1 and p-HS1 than healthy subjects, and the specific inhibitors of Src and Syk , PP2 and piceatannol, inhibited the increase in p-HS1 in LPS-induced cell model.Conclusions Function of platelet is closely related to HS1 in sepsis and it will be a target for sepsis therapy.
7.The research of IMRT plan optimization on dose limitation of sub regional auditory organ
Ming GUO ; Shengzi WAMG ; Zhaoxia WU ; Changming MA ; Lei WANG ; Xiaochen NI ; Feng ZHU
Chinese Journal of Radiation Oncology 2015;(4):438-441,442
Objective To investigate the effects of reducing the auditory organ dose by limitation of sub regional auditory organ in IMRT plan. Methods Total 223 cases of nasopharyngeal carcinoma patients were divided into group A and group B. In group A, the IMRT plans of 114 patients were designed by limiting overall auditory organ dose. In group B, the IMRT plans of 109 patients were designed by limiting sub regional auditory organ dose. According to the Clinical prescription, the IMRT plans were designed. Paried t?test was difference between groups. Results By comparing the two groups of auditory organ dose, in all stages, the tympanic cavity Dmean average in group B decreased by T1 vs. 17?? 7%,T2 vs. 22?? 4%,T3 vs. 15?? 7% and T4 vs. 14?? 2% ( P= 0?? 000,0?? 000,0?? 000,0?? 000);cochlea Dmean average decreased by T1 vs. 11?? 0%, T2 vs. 20?? 1%, T3 vs. 10?? 0% and T4 vs. 9?? 0%(P= 0?? 004,0?? 000,0?? 007,0?? 036);vestibule Dmean average decreased by T1 vs. 22?? 6%, T2 vs. 31?? 8%, T3 vs. 20?? 6% and T4 vs. 21?? 4%, significantly less than in group A (P= 0?? 000,0?? 000,0?? 000,0?? 000). The bony portion of eustachian tube Dmean average in group B decreased were not significantly less than in group A (decreased by 3?? 4%,6?? 8%,3?? 6%,0?? 1%;P= 0?? 291, 0?? 006,0?? 155,0?? 963). Conclusions In IMRT plan, optimization on dose limitation of sub regional auditory organs were used to reduce the auditory organ dose and decrease the radiation damage to auditory organ.
8.Generation and characterization of monoclonal antibody against glycoprotein VP1 encoded by caprine enterovirus CEV-JL14
Haibing LU ; Mingyue WANG ; Lisai ZHU ; Yajing LIU ; Changming GUO ; Qun ZHANG ; Yue YUAN ; Changchun TU ; Xinping WANG
Chinese Journal of Veterinary Science 2017;37(8):1468-1472
VP1 gene encoded by the newly identified caprine enterovirus CEV-JL 14 was amplified and cloned to prokaryotic expression vector pGEX-4T-1.Recombinant GST-VP1 fusion protein was expressed,purified,emulsified with Freund's complete adjuvant and used to immunize the BALB/c mice following a standard procedure.The spleen cells from immunized mice were collected and fused with myeloma cells after its antibody titer reached over 104 times detected by indirect ELISA.Hybridoma cell clones secreting monoclonal antibodies against VP1 were screened and their stability and specificity were further determined.The identified hybridoma cells were injected to mice intraperitoneally and ascites were collected at 7 DPI.Isotypes of the monoclonal antibodies against the recombinant VP1 protein were characterized to be either IgG1 or IgG2b,which showed a high specificity for detection of caprine enterovirus antigens by immunoperoxidase monolayer assay,thus laying a solid basis for future study related to viral pathogenesis,detection and diagnostics for caprine enterovirus infection.
9.Meta-analysis of the efficacy of recombinant human growth hormone and human serum albumin in the treatment of liver cirrhosis with hypoproteinemia
Yaochang GUO ; Changming JIANG ; Wenlin SHU
Chinese Journal of Primary Medicine and Pharmacy 2020;27(23):2825-2829
Objective:To evaluate the efficacy, safety and economy of recombinant human growth hormone (rhGH) in the treatment of patients with liver cirrhosis and hypoproteinemia.Methods:Computer search for randomized controlled trials included in Chinese Biomedical Literature Database(CBM), Wanfang Medicine, Weipu Medicine, Embase, PubMed, Cochrane Library and other databases from January 1, 2000 to January 1, 2019, and excluded according to certain criteria.RevMan5.3 software was used for statistical analysis.Results:A total of 13 articles were included in the meta-analysis, a total of 739 patients.The results showed that the short-term efficacy of rhGH in the treatment of cirrhosis with hypoproteinemia had no statistically significant difference compared with human serum albumin (HSA)[WMD=-0.42, 95% CI(-1.77, 0.92), P=0.54], but the mid-and long-term effects of rhGH were better[WMD=3.42, 95% CI(0.13, 6.70), P=0.04; WMD=7.12, 95% CI(4.75, 9.50), P<0.001]. Conclusion:The application of rhGH in the treatment of cirrhosis with hypoproteinemia is equivalent to the infusion of HSA in short-term efficacy, but the duration of treatment is prolonged, has no obvious adverse reactions, and is more economical.
10.Circular RNA HECTD1 participates in oxygen-glucose deprivation-induced neuronal cell damage by regulating miR-98-5p/ephrin A4 expressions
Yifeng LIU ; Changming WEN ; Chuanjie WU ; Jun GAO ; Jun SUN ; Meng SUN ; Shiqian GUO
Chinese Journal of Neuromedicine 2022;21(6):541-552
Objective:To explore whether circular RNA HECTD1 (circ-HECTD1) is involved in oxygen-glucose deprivation (OGD)-induced neuronal cell damage by regulating the expressions of miR-98-5p/ephrin A4 (EPHA4).Methods:Mouse primary cortical neuronal cells were isolated and cultured in vitro. The targeting relations of circ-HECTD1 and miR-98-5p with EPHA4 were detected by dual luciferase reporter assay and RNA binding protein immunoprecipitation assay. These neurons were randomly divided into control group (cultured for 24 h under normal condition) and 6, 12 and 24 h OGD treatment groups (treated with OGD for 6, 12 and 24 h, respectively), OGD+Vector group and OGD+circ-HECTD1 group, OGD+small interfering RNA (siRNA) negative control (si-NC) group and OGD+siRNA circ-HECTD1 (si-circ-HECTD1) group, OGD+micro RNA (miR) negative control (miRNC) group and OGD+miR-98-5p mimic group, OGD+miRNA inhibitor negative control (anti-miRNC) group and OGD+miR-98-5p inhibitor (anti-miR-98-5p) group, OGD+miR-98-5p mimic+pcDNA group and OGD+miR-98-5p mimic+EPHA4 group, OGD+si-circ-HECTD1+anti-miR-NC group and OGD+si-circ-HECTD1+miR-98-5p inhibitor group; pCD5-ciR empty vector, pCD5-ciR-circ-HECTD1, si-NC, si-circ-HECTD1, miR-NC, miR-98-5p mimic, anti-miR-NC or anti-miR-98-5p were transfected into the neurons, and miR-98-5p mimi and pcDNA3.1 empty vector, miR-98-5p mimic and pcDNA3.1-EPHA4 overexpression vector, si-circ-HECTD1 and anti-miR-NC, or si-circ-HECTD1 and anti-miR-98-5p were co-transfected into the neurons. After 24 h of OGD treatment, the circ-HECTD1, miR-98-5p and EPHA4 mRNA expressions were detected by real-time fluorescent quantitative PCR (qRT-PCR), the EPHA4 protein expression was detected by Western blotting, the proliferation activity was detected by MTT assay, the apoptosis rate was detected by flow cytometry, the levels of interleukin (IL)-1β and tumor necrosis factor (TNF)-α in cell culture medium were detected by ELISA, and the activities of superoxide dismutase (SOD) and malondialdehyde (MDA) were detected by kit assay. Results:(1) Targeting relations between circ-HECTD1 and miR-98-5p, and EPHA4 and miR-98-5p were verified. (2) As compared with the control group, the neurons in 6, 12 and 24 h OGD treatment groups had significantly increased circ-HECTD1 and EPHA4 protein expressions and significantly decreased miR-98-5p expression ( P<0.05). (3) As compared with OGD+Vector group, OGD+circ-HECTD1 group had significantly increased circ-HECTD1 expression, and significantly decreased miR-98-5p expression ( P<0.05); as compared with OGD+si-NC group, OGD+si-circ-HECTD1 group had significantly increased miR-98-5p expression, and significantly decreased EPHA4 mRNA and protein expressions ( P<0.05); as compared with OGD+miR-NC group, OGD+miR-98-5p mimic group had significantly increased miR-98-5p expression, and significantly decreased EPHA4 protein expression ( P<0.05); as compared with OGD+anti-miR-NC group, OGD+anti-miR-98-5p group had significantly decreased miR-98-5p expression, and significantly increased EPHA4 protein expression ( P<0.05); as compared with the OGD+si-circ-HECTD1+anti-miR-NC group, OGD+si-circ-HECTD1+anti-miR-98-5p group had significantly increased EPHA4 mRNA and protein expressions ( P<0.05). (4) As compared with the control group, the OGD groups had significantly decreased cell viability and SOD activity, and significantly increased IL-1β and TNF-α levels, apoptosis rate and MDA activity ( P<0.05); as compared with the OGD+si-NC group, the OGD+si-circ-HECTD1 group had significantly decreased cell apoptosis rate, IL-1β and TNF-α levels, and MDA activity, and significantly increased cell viability and SOD activity ( P<0.05); as compared with the OGD+si-circ-HECTD1+anti-miR-NC group, the OGD+si-circ-HECTD1+anti-miR-98-5p group had significantly decreased cell viability and SOD activity, and significantly increased IL-1β and TNF-α levels, apoptosis rate and MDA activity ( P<0.05); as compared with the OGD+miR-NC group, OGD+miR-98-5p mimic group had significantly decreased cell apoptosis rate, IL-1β and TNF-α levels, and MDA activity, and significantly increased cell viability and SOD activity ( P<0.05); as compared with OGD+miR-98-5p mimic+pcDNA group, OGD+miR-98-5p mimic+EPHA4 group has significantly increased cell apoptosis rate, IL-1β and TNF-α levels, and MDA activity, and significantly increased cell viability and SOD activity ( P<0.05). Conclusion:Knockdown of circ-HECTD1 could ameliorate the OGD-induced neuronal cell damage in mice by targeting the expressions of miR-98-5p/EPHA4.