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ObjectiveTo investigate the effect of Tangzhi pills on the improvement of insulin resistance （IR） in the liver with type 2 diabetes （T2DM） by regulating phosphatidylinositol 3-kinase （PI3K）/protein kinase B （Akt） signaling pathway based on differential genes and its possible molecular mechanism. MethodT2DM rat models were prepared by high fat （HFD） diet combined with streptozotocin （STZ） intraperitoneal injection. The experiment was divided into blank group， model group， metformin hydrochloride group （0.18 g·kg-1）， Tangzhi pills high （1.08 g·kg-1）， medium （0.54 g·kg-1） and low （0.27 g·kg-1） dose groups. Rat serum， liver， and pancreatic tissue were collected， and the pathological tissue of the liver and pancreas was observed using hematoxylin-eosin （HE） staining. The fasting blood glucose level （FBG） was detected， and oral glucose tolerance （OGTT） tests were conducted. Enzyme-linked immunosorbent assay （ELISA） was used to detect fasting serum insulin （FINS） and glycated hemoglobin （GHb） levels in rats. IR homeostasis model index （HOMA-IR）， β cellular homeostasis index （HOMA-β）， and insulin sensitivity index （ISI） were calculated. Biochemical methods were used to determine the levels of triglyceride （TG）， total cholesterol （TC）， low-density lipoprotein （LDL-C）， and high-density lipoprotein （HDL-C） in rat serum. Transcriptomics obtained differentially expressed mRNA from liver tissue and enriched differentially expressed pathways. Real-time reverse transcriptase polymerase chain reaction （Real-time PCR） was used to detect the mRNA expression of cyclic adenylate responsive element binding protein 3-like protein 2 antibody （CREB3l2）， B-lymphocyte tumor 2 （Bcl-2）， Toll-like receptor 2 （TLR2）， cyclin-dependent kinase inhibitor 1A （CDNK1A）， and DNA damage induced transcription factor 4-like protein （DDIT4） in liver tissue. Western blot was used to detect the protein expression of phosphorylated phosphatidylinositol 3-kinase （p-PI3K）， phosphorylated protein kinase B （p-Akt）， glucose transporter 4 （GLUT4）， insulin receptor （INSR）， and insulin receptor substrate 2 （IRS2）. ResultThe pharmacodynamic experiment results showed that compared with model group， Tangzhi pills groups repaired liver and pancreatic tissue to varying degrees， reduced blood sugar （P<0.01）， and promoted a decrease in serum FINS， GHb， and HOMA-IR （P<0.05， P<0.01）. In addition， HOMA-β and ISI increased （P<0.05， P<0.01）. The levels of TC， TG， and LDL-C decreased （P<0.05， P<0.01）， while the levels of HDL-C increased （P<0.05， P<0.01）. The transcriptomics experimental results confirmed that the PI3K/Akt signaling pathway was significantly expressed in both the blank group and model group， as well as in the high-dose Tangzhi pills group and model group. CDNK1A， DDIT4， CREB3l2， Bcl-2， and TLR2 were significantly differentially expressed mRNA during TG intervention in T2DM. Compared with the model group， the protein expression of p-PI3K， p-Akt， GLUT4， INSR， and IRS2 increased in all Tangzhi pills groups （P<0.01）. The mRNA expression of CREB3l2， Bcl-2， and TLR2 increased （P<0.01）， while that of CDNK1A and DDIT4 decreased （P<0.01）. ConclusionTangzhi pills may regulate the PI3K/Akt signaling pathway based on the differential mRNA expression of CREB3l2， Bcl-2， TLR2， CDNK1A， and DDIT4， thereby improving IR in the liver with T2DM.
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Objective To explore the effect of single level lumbar disc herniation on the morphology and diameter of sciatic nerve,and to identify the correlation between the diameter change in sciatic nerve and severity of clinical symptoms,as well as the prognosis of surgical treatment in patients with lumbar disc herniation.Methods From January 1,2017 to December 31,2017,Seventy lumbar disc herniation (LDH) patients who underwent single-level posterior lumbar spine surgery were recruited in this retrospective analysis study.Specific data including age,gender,the level of a disc herniation,the type of disc herniation,symptomatic side,surgical procedure were recorded respectively.In addition the morphological changes and the diameter of bilateral sciatic nerve were recorded preoperatively and postoperatively by B-mode ultrasound.Furthermore,the pain in patients and neurological function were evaluated by visual analogue scale (VAS,back pain and leg pain),Japanese orthopaedic association scores-lumbar (JOA) and Oswestry disability index (ODI).The correlation between the diameter of sciatic nerve and clinical features,as well as clinical prognosis of patients were identified in the present study.Results For patients with acute lumbar disc herniation,the diameter of sciatic nerve in affected side was 5.19±1.03 ram,which is significantly higher than that in the unaffected side (4.57±0.64 mm,t=6.735,P=0.000).In addition,preoperative ratio of the affected side to the healthy side of the sciatic nerve showed strong correlation with the VAS of leg pain (r=0.838,P=0.001),JOA (r=-0.857,P=0.001),and ODI score(r=0.881,P=0.000),but not with the VAS of back pain (r=-0.061,P=0.614).Three months after surgery,the diameter of sciatic nerve in the affected side decreased to 4.58±0.63 mm (t=6.865,P=0.000),while the unaffected side showed no significant change(t=0.300,P=0.765).Clinical improvement was observed in all the patients postoperatively.The changes in the diameter of sciatic nerve postoperatively in affected side showed strong correlation to the rate of improvement in VAS of leg (r=0.624,P=0.003),JOA(r=0.615,P=0.003) and ODI scores (r=0.722,P=0.002),but not to the rate of improvement in VAS of back (r=-0.025,P=0.836).Conclusion Single root compression in patients with single level disc herniation might cause morphological changes such as thickening and edema in sciatic nerve,which were closely related to the severity of clinical symptoms and the prognosis of surgical treatment.
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Objective To study the clinical effect of sevoflurane combined with remifentanil in laparoscopic cholecystectomy (LC).Methods50 cases of laparoscopic cholecystectomy in Tianjin Haihe Hospital from February 2015 to September 2016 were selected as the research object, and randomly divided into the control group and the experimental group, each group had a total of 25 cases.The patients in control group were treated with continuous intravenous infusion of propofol.The experimental group received sevoflurane.Patients in the experimental group and the control group were given remifentanil after surgery.Compared with the experimental group and control group of patients with the degree of recovery and related Linchuan indicators.ResultsAfter the corresponding anesthesia, the control group of patients with recovery time was (13.4±3.1) minutes, the score was (4.1±1.2) points, extubation time was (14.6±3.7) minutes.However, the extubation time and recovery time of the control group were significantly longer than those of the experimental group, and the recovery score was lower than that of the experimental group, with statistical difference (P<0.05).There was no obvious adverse reaction in the experimental group and the control group, and there was no significant difference in the changes of cardiovascular factors.ConclusionSevoflurane combined with remifentanil for laparoscopic cholecystectomy can improve the anesthetic effect to a great extent, stable operation and postoperative patients with cardiovascular function, recovery time is short, the clinical significance of only one step promotion and application.
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Objective We used the DNA methylation microarrays to investigate the differential methylation genes and loci sites in Kashin-Beck disease (KBD),to study the relationship between DNA methylation and KBD pathogenesis.Methods Totally 12 KBD adults and 12 healthy adults were selected and peripheral blood samples were collected and DNA was extracted.Illumina 450K bead-chip was applied to detect methylation status in KBD and healthy controls.Aberrant hyper-methylated sites were filtrated according to the P value after correction and methylation differences,together with GenomeStudio soft.Screened genes were validated using bisulfite sequencing polymerase chain reaction (BSP) technology.Results A total of 484 948 loci sites were analyzed and compared,93 differential methylated loci were found by comparing KBD and normal people,including 34 hypermethylated sites and 59 hypomethylated sites.There were 50 genes corresponding to the loci,43 genes not reported in literature.According to gene ontology analysis,the genes were involved in the immune response,antigen processing,phosphate and phosphoric acid metabolism and phosphorylation and the process of metal ions in combination.However,in the verification test using BSP method,there was no significant difference in methylation rate in human leukocyte antigen (HLA)-DRB1 between the case and the control group (48％ vs 70％,x2 =3.688,P ＞ 0.05).Conclusions The high and low differentially methylated sites in peripheral blood DNA of KBD patients are significantly different from those of the health control.HLA-DRB1 locus is not significantly different between the BSP verification test and methylation chip.
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Objective To investigate the clinical efficacy of the self-retaining laryngeal microsurgical operation in the treatment of Reinke ’ s edema of vocal cords . Methods There were 24 cases of Reinke ’ s edema who were treated with self-retaining laryngoscope mucosal stripping surgery of vocal cords from January 2004 to December 2009 ( traditional group ) , while another group of 32 cases of Reinke ’ s edema were treated with self-retaining laryngoscopic microsurgery lateral submucosal incision micro-flap operation of vocal cords from January 2010 to December 2015 ( laryngeal microsurgery group ) .The two groups were executed with electronic laryngoscopy and subjective voice evaluation GRBAS ( The Speech and Language Institute of Japanese in 1979, G: Grade, R:Roughness, B:Breathness, A:Asthenia, S:Strain) in pre-operation and post-operation (1 week, 3 weeks, 8 weeks).The wound healing time of vocal cords , hoarseness improved time and voice improvement were retrospectively compared between the two groups . Results The patients in laryngeal microsurgery group had earlier voice hoarse improvement and more rapid mucosal epithelial of vocal cords recovery time in post-operation as compared with the patients in the traditional group [voice improving time, (7.3 ±1.9) d vs. (11.3 ±2.7) d, t=-6.481, P=0.000;mucosal epithelial of vocal cords recovery time , (12.2 ±3.1) d vs.(20.1 ±3.4) d, t=-9.062, P=0.000].The results of pre-operative voice evaluation with GRBAS showed no significant differences between the two groups, but the results of post-operative evaluation of each session (1 week, 3 weeks, 8 weeks) showed that the main data were statistically different.The GRBAS score of laryngeal microsurgery group was lower than that of traditional group in post -operation, especially after 8 weeks (8 weeks after post-operation, G:1.0 ±0.8 vs.1.6 ±0.2, t=-3.584, P=0.000; R:1.0 ±0.9 vs. 1.5 ±0.4, t=-2.536, P=0.014;B:1.0 ±0.6 vs.1.4 ±0.5, t=-2.647, P=0.011).In laryngeal microsurgery group, 24 cases were cured , 5 cases were effective and 3 cases were invalid , contrasting in the traditional group with 11 cases of cured , 7 cases of effective and 6 cases of invalid respectively, with a significant difference (Z=-2.239, P=0.025).No significant difference in effective rate between the two groups [90.6%(29/32) vs.75.0%(18/24),χ2 =1.459, P=0.227]. Conclusion The clinical efficacy of self-retaining laryngoscope microsurgery in the treatment of Reinke ’ s edema of vocal cords is faster and better comparing traditional operation , with a more significant pronunciation quality improvement .
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AIM:To determine the biological feature of circulating endothelial cells (CECs) in acute promye-locytic leukemia ( APL) patients before and after treatment , and to analyze the relationship between CECs and the clinical characteristics .METHODS: The CECs were sorted from peripheral blood by magnetic-activated cell sorting and then counted by 3-color flow cytometry.The cells were identified by immunofluorescence staining for the expression of CD 146, CD31, CD144, VEGFR-2, CD45 and CD133.The CECs were cultured in vitro, and the tube formation and colony-forming rate were determined .RESULTS:Increased quantity of CECs was observed in CD 34 positive group and group with WBC >10 ×109/L (P<0.05).The quantity of CECs had a significant difference among low risk , medium risk and high risk groups (P<0.05).The positive rate of CD133 and quantity of CECs significantly reduced in 32 APL patients when they gain complete remission after treatment (P<0.05).The amount of tube formation and colony-forming rate were significant-ly reduced after treatment (P<0.05).The ratio of CECs quantity from APL patients after treatment to that before treatment had a negative correlation with arsenic concentration in urine on day 7 during As2O3 treatment (P<0.05).CONCLU-SION:Accurately counting CECs may be helpful for evaluating prognosis and designing treatment strategy .
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Objective To compare cardiovascular responses of intubations by Discopo and Macintosh laryngoscope in elderly patients with hypertension. Methods Sixty elderly hypertensive patients with ASAⅡ~Ⅲundergoing elective surgery were equally randomized into two groups. After induction of general anesthesia , orotracheal intubation was performed with Discopo or Macintosh. The Mean arterial pressure (MBP) and heart rate (HR), as while as Electrocardiogram (ECG) and Blood oxygen saturation (SpO2), were recorded before (baseline values) and at 1,3,5 minutes after intubation(post-induction values). The rate of one-time successive intubation,intubation time and complication were also recorded. Results Compared with the macintosh group, the rate of one-time successive intubation was higher and the intubation time was shorter in the Discopo group (P< 0.05, respectively),with no complications. Compared with T0,MBP and HR at T2 in the two groups and T3 in the Macintosh group increased significantly (P < 0.05). Compared with the Macintosh group,MBP and HR at T2,T3 of the Discopo group decreased significantly (P < 0.05). Conclusion Little effect of tracheal intubation with Discopo on cardiovascular response was observed in elderly hypertensive patients.
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Objective To investigate the changes in the expression of matrix metalloproteinase-1 (MMP-1),matrix metalloproteinase-3 (MMP-3) and tissue inhibitor of metalloproteinase-3 (TIMP-3) in the articular cartilage of patients with Kashin-Beck disease (KBD) and the role of these proteins in pathogenesis of KBD.Methods The postoperative cartilage samples were collected from patients with KBD,osteoarthritis and patients with non-bone disease (control).The expression of MMP-1,MMP-3 and TIMP-3 in the cartilage was detected by immuohistochemistry,and the positive chondrocytes were counted in different layers of the articular cartilage under microscope.Results The positive rates of MMP-1 in the upper [(67.00 ± 1.69)％] and deeper [(22.07 ± 29.66)％] layers of articular cartilage from patients with KBD,and in the deeper layer of articular cartilage from patients with osteoarthritis [(70.52 ± 37.84)％] were significantly higher than those in the control group [(51.73 ± 36.74)％,(3.75 ± 6.85)％,all P ＜ 0.05].The positive rates of MMP-3 in the deeper layer of articular cartilage from patients with KBD [(28.84 ± 31.13)％] and in the middle and deeper layers of articular cartilage from patients with osteoarthritis [(55.69 ± 35.00)％,(45.96 ± 35.38％)] were significantly higher than those in normal cartilage [(34.09 ± 28.54)％,(14.46 ± 18.32)％,all P ＜ 0.05].The positive rates of TIMP-3 in the middle layer of articular cartilage from patients with KBD [(21.25 ± 25.23)％] and in the middle and deeper layers of articular cartilage from patients with osteoarthritis [(20.40 ± 22.19)％,(18.10 ± 22.58)％] were significantly lower than those in normal cartilage [(36.74 ± 26.61)％,(7.81 ± 20.58)％,all P ＜ 0.05].Conclusion MMP-1,MMP-3 and TIMP-3 play important roles in the articular cartilage damage of KBD.
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This study was aimed to simultaneously determinate the content of naringin, hesperidin and atractylodin in Y un-Pi (YP) Powder extract by HPLC. A Thermo ODS-2 column (250 mm í 4.6 mm, 5 μm) was used for the gra-dient elution with a methanol-water of 0~5 min (30%~40% A), 5~15 min (40%~50% A), 15~20 min (50%~79%A), 20~30 min (79%~100% A), 30~45 min (100% A), 45~55 min (100%~30% A), 55~60 min (30% A). The col-umn temperature was at 30℃. The detection wavelength was at 283 nm. The flow rate was 1 mL·min-1. The linear range was determined to be 0.009 16~0.137 00 μg (r = 0.999 9), 0.006 22~0.091 30 μg (r = 0.999 8), 0.003 58~0.053 70 μg (r = 0.999 9), respectively for naringin, hesperidin and atractylodin. The average recovery rate of pterodontic acid was 103.51%, 96.82% and 97.65%. And RSD was 1.37%, 1.41%, 1.49% (n = 6). It was concluded that the method had good specificity and repeatability. It was considered as a good tool for the content determination of naringin, hesperidin and atractylodin in YP Powder extract.
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Objective To investigate the influences of polyethylene glycol on the solubility and in vitro dissolution of m-nisoldipine,which could provide guidance for chosing formulations of m-nisoldipine. Methods Solid dispersions of m-nisoldipine were prepared by solvent-melting method with polyethylene glycol6000 matrix. DSC and XRD spectroscopy were applied to identify the solid dispersions. The solubility and in vitro dissolution were detected by UV spectroscopy. Results The DSC and XRD map were different from the crude drug and their physical mixtures. The dissolution rates(13,15,17) were faster(35. 31%,38. 71%,41. 48%) than that of the crude drug(26. 80%),and the dissolution rates of the solid dispersions in the same ratio were higher than the physical mixtures. Conclusion DSC analysis indicated that eutectic compounds were produced by the m-nisoldipine and polyethylene glycol,in which polyethylene glycol6000 acts as a carrier. The solubility and in vitro dissolution of m-nisoldipine can be increased.
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This study was aimed to screen out an optimal pharmaceutical formulation of Wuji Gastric Floating Sus-tained-release Tablets. With multicomponents of release for index, the extra-floating ability of the tablets was stud-ied with the orthogonal test of four excipients, which were HPMC (K100M), PEG 6000, sodium bicarbonate and se-tanol. The results showed that the combined optimal pharmaceutical formulation was 35% HPMC, 5% PEG 6000, 10% sodium bicarbonate, 15% setanol, 10% MCC and 25% main drug. It was concluded that Wuji Gastric Floating Sustained-release Tablets had the characteristics of slow-release and long floating duration which comply with re-quirements of formulation design. This preparation technique is with good maneuverability.
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This article was aimed to study the multicomponent in vivo releasing rules of Zuojin Gastric Floating Sus-tained-release Tablet. Stomach contents of rabbits were collected at different times and then prepared into solutions for the study. The HPLC was used to establish fingerprints of each time and fingerprints of completely dissolved samples. The in vivo releasing rates were calculated. The evaluation was made on releasing behaviors among index components as well as between index components and preparations. The results showed that within 10 h, the f2 of berberine and limonin, rutaecarpine, preparation were 52, 61.5, 66.4, respectively. The f2 of limonin and rutae-carpine, preparation were 70, 46.5, 53.7, respectively. It was concluded that within 10 h, the releasing behavior of berberine and limonin, rutaecarpine, preparation was similar. The releasing behavior of rutaecarpine and limonin, preparation was similar. However, the releasing behavior of limonin and preparation was different.
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Objective To characterize the function of human anti-BP180 single-chain Fv antibody (scFv) in vitro. Methods The IgG autoantibodies against BP180 were purified by affinity chromatography from the sera of patients with BP. The inhibitive effect of previously constructed anti-BP180 scFv on the binding of anti-BP180 IgG autoantibodies to the recombinant NC16A domain of human BP180 antigen was observed by competitive ELISA, competitive immunofluorescence assay and competitive inhibition test for complement activation. Results As ELISA revealed, the scFv significantly inhibited the binding of anti-BP180 IgG autoantibodies to the corresponding antigen (P ＜ 0.01 ), and the inhibitive effect was dose-dependent within the concentration range from 0 to 60 μg/ml. The inhibitive rate peaked at 69.50%. The deposition of anti-BP180-IgG autoantibodies in basement membrane zone and the IgG autoantibody-mediated complement C3 activation were both suppressed by the scFv of 40 μg/ml. Conclusion The genetically engineered anti-BP180 scFv has a certain inhibitive effect on the binding of BP-IgG autoantibodies to BP180 antigen and on the subsequent complement activation in vitro.
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Objective To detect the proliferation of and production of interferon-γ by drug-specific peripheral T cells from patients with severe drug eruption.Methods Peripheral blood mononuclear cells (PBMCs) were isolated from 10 patients with severe drug eruption,10 patients with mild or moderate drug eruption and 10 normal human controls,stimulated with causative drugs to obtain drug-specific T cells.Then,both PBMCs and drug-specific T cells were stimulated with causative drugs or unrelated drugs followed by the detection of secretion levels of IFN-γ with ex vivo enzyme-linked immunodotting (ELISpot) assay and cultured ELlSpot assav respectively.Results After stimulation with causative drugs,a higher level of IFN-γ was secreted by PBMCs and drug-specific T cells from patients with severe drug eruption compared with those from normal human controls (both P<0.01).and by drug-specific T cells than by PBMCs (P<0.01).The culture with unrelated drugs could neither induce the generation of drug-specific T cells nor promote the secretion of IFN-γ by PBMCs from the patients.Drug-specific T cells still existed in the peripheral blood of 3 patients within 1 to 3 years after recovery of drug eruption.Conclusions There are drug-specific T cells in peripheral blood of patients with severe drug eruption,and they may persist for a certain period of time after recovery of drug eruption.Ex vivo ELISpot combined with cultured ELISpot may be applied to the identification of causative drugs in vivo.
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Objective QR-DVT 9000 NEWTOM CT, an oral maxillofacial dedicated CT, was applied in the diagnosis of maxillary sinus related diseases. Its value and technique are summarized. Method Patients with maxillary sinus related diseases were scanned and images were reconstructed with QR-DVT 9000 CT. The axial view of reconstructed image was analyzed in sagittal plane, coronal plane and 3-D. Results QR-DVT 9000 NEWTOM CT could display the position, shape and anatomic relationship of maxillary sinus with adjacent structures in sagittal image, coronal image and 3-D image. Conclusion QR-DVT 9000 NEWTOM CT, an oral maxillofacial dedicated CT, is considerably valuable in the diagnosis of maxillary sinus related diseases, and it can guide clinical surgery more effectively than routine X ray examination.
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AIM: To establish the method of determining astragaloside Ⅳ and salidroside in Zhenqi Fuzheng Capsule(Fructus Ligustri Lucidi,Radix Astragali,etc.). METHODS: Astragaloside Ⅳ was determined by HPLC-ELSD on Inertsil ODS-3 C_(18) column(250 mm?4.6 mm,5 ?m) using a mobile phase of acetonitrile-water(37∶63).The flow rate was 1.0 mL/min and detector parameters were set as follows: drift tube temprature was at (106 ?C);carrier gas(air) flow rate was 2.7 L/min.Salidroside was determined by RP-HPLC on the same column using a mobile phase composed of acetonitrile-water with gradient elution.The detection wavelength was at 278 nm and the flow rate was 1.0 mL/min. RESULTS: Calibration curve of astragaloside Ⅳ was linear between 0.76 ?g and 3.8 ?g under the chromatographic condition,R=0.999 5.The average recovery of astragaloside Ⅳ was(98.4%).RSD was 0.9%(n=6).And the calibration curve of Salidroside was linear between 0.4 ?g and 4 ?g,R=0.999 9.The average recovery of salidroside was 100.7%.RSD was 2.1%(n=5). CONCLUSION: These methods were simple,accurate and sensitive,so it can be used for the quality control of Zhenqi Fuzheng Capsule.