Identification of components in vitro and plasma of Jinhua Qinggan granules and preliminary pharmacokinetics analysis / 药学学报

As one of the "Three Drugs Three Prescriptions" anti-COVID-19 traditional Chinese medicine, Jinhua Qinggan granules (JHQG) has been proved to have clear clinical effects. With complex medicinal flavors and ingredients, there is no systematic research report on chemical composition in vivo or in vitro. An ultrahigh pressure liquid chromatography-quadrupole-time of flight mass spectrometry (UPLC-QTOF/MS) method was developed in this study to identify the components of the anti-COVID-19 traditional Chinese medicine JHQG granules. Analyze the collected rat plasma samples after administration and explore the exposed components in rats within 8 hours after intragastric administration. Preliminary pharmacokinetic analysis was then performed on this basis. Through UPLC-QTOF/MS analysis and verification by standard products, a total of 77 chemical components in JHQG formula have been identified, among which 22 compounds were highly exposed in vivo, mainly derived from three medicinal materials of honeysuckle, scutellaria and forsythia. Through the assessment of the blood drug concentration by the compartment model, 6 PK parameters of 4 high-exposure chemical components have been obtained, clarifying the metabolic characteristics of the main exposed components in JHQG briefly. The method is simple, efficient, sensitive and accurate and provides research basis to the clarification of the pharmacodynamics material basis and mechanism of JHQG, which has certain reference significance for the basics and applications research of the traditional Chinese medicine prescriptions in fighting the SARS-CoV-2.

Study of Adverse Events of Endotracheal Intubation / 中国医疗器械杂志

OBJECTIVE@#In order to further decrease and reduce the serious adverse events of silicone rubber endotracheal intubation in clinical use, especially in anesthesia and intensive care.@*METHODS@#Through the first stage analysis on the registration and certification of endotracheal intubation products in China, adverse events of products in recent five years in Zhejiang province, domestic and foreign literature of adverse events of products, retrieval of product citation standards, content integrity of product instructions, and expert seminar on serious adverse events, combined with the air leakage of endotracheal intubation products in recent two years, product material and clinical application with normative aspects.@*RESULTS@#Silicone rubber endotracheal intubation products in clinical intensive care have certain clinical safety risks, especially for long-term use of critically ill patients.@*CONCLUSIONS@#According to the four cases of serious adverse events of silicone rubber endotracheal intubation in the clinical intensive care unit, we put forward some suggestions for the manufacturers, clinical users and regulatory agencies to further decrease and reduce the serious adverse events of silicone rubber endotracheal intubation.

Curcumin Reverses 5-Fluorouracil Resistance by Promoting Human Colon Cancer HCT-8/5-FU Cell Apoptosis and Down-regulating Heat Shock Protein 27 and P-Glycoprotein / 中国结合医学杂志

OBJECTIVE@#To investigate the potential mechanisms that curcumin reverses 5-fluorouracil (5-FU) multidrug resistance (MDR).@*METHODS@#Cell growth and the inhibitory rate of curcumin (2-25 μg/mL) and/or 5-FU (0.05-1000 μg/mL) on human colon cancer HCT-8 and HCT-8/5-FU (5-FU-resistant cell line) were determined using cell counting kit-8 (CCK-8) assay. Apoptosis and cell cycle after 5-FU and/or curcumin treatment were detected by flow cytometry (FCM) and transmission electron microscopy (TEM). The expression of the multidrug resistance related factors p-glycoprotein (P-gp) and heat shock protein 27 (HSP-27) genes and proteins were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and Western blotting (WB), respectively.@*RESULTS@#The inhibitory rate of curcumin or 5-FU on HCT-8 and HCT-8/5-FU cells proliferation at exponential phase were in a dosedependent manner, HCT-8 cell line was more sensitive to curcumin or 5-FU when compared the inhibitory rate of HCT-8/5-FU. The 50% inhibitory concentration (IC) of combination 5-FU and curcumin (4.0 μg/mL) in HCT-8/5-FU was calculated as 179.26 μg/mL, with reversal fold of 1.85. Another IC of combination 5-FU and curcumin (5.5 μg/mL) in HCT-8/5-FU was calculated as 89.25 μg/mL, with reversal fold of 3.71. Synergistic effect of 5-FU and curcumin on HCT-8 and HCT-8/5-FU cells were found. The cell cycle analysis performed by FCM showed that HCT-8 and HCT-8/5-FU cells mostly accumulated at G/G phase, which suggested a synergistic effect of curcumin and 5-FU to induce apoptosis. FCM analysis found that the percentage of apoptosis of cells treated with curcumin, 5-FU and their combination were significantly increased compared to the control group (P<0.05), and the percentage of apoptosis of the combination groups were slightly higher than other groups (P<0.05). The mRNA levels of P-gp (0.28±0.02) and HSP-27 (0.28±0.09) in HCT-8/5-FU cells treated with combination drugs were lower than cells treated with 5-FU alone (P-gp, 0.48±0.07, P=0.009; HSP-27, 0.57±0.10, P=0.007). The protein levels of P-gp (0.25±0.06) and HSP-27 (0.09±0.02) in HCT-8/5-FU cells treated with combination drugs were decreased when compared to 5-FU alone (P-gp, 0.46±0.02, P=0.005; HSP-27, 0.43±0.01, P=0.000).@*CONCLUSIONS@#Curcumin can inhibit the proliferation of human colon cancer cells. Curcumin has the ability of reversal effects on the multidrug resistance of human colon cancer cells lines HCT-8/5-FU. Down-regulation of P-gp and HSP-27 may be the mechanism of curcumin reversing the drug resistance of HCT-8/5-FU to 5-FU.

Role of cytoskeleton in autophagy / 生物医学工程学杂志

Cell autophagy plays a key role in maintaining intracellular nutritional homeostasis during starvation through elimination of aberrant or obsolete cellular structures. The cellular cytoskeleton has a crucial role in multiple processes involving membrane rearrangements and vesicle-mediated events. Autophagy is mediated by both microtubules and actin networks: microtubules promote the synthesis of autophagosome and are related to the movement of autophagosome; actin networks have been implicated in structurally supporting the expanding of phagophore, moving autophagosomes and enabling their efficient fusion with the lysosome; non-muscle myosinⅡoperates in the early stages of autophagy during the initiation and expansion of the phagophore, whereas myosinⅥ and myosin 1C are involved in the late stages of autophagosome maturation and fusion with the lysosome, respectively. This review summarizes the multiple regulation of cytoskeleton on autophagy and focuses on the regulation of autophagy by actin and myosin, providing a new approach for the study of pathogenesis and innovative therapies of autophagy related diseases.

Clinical effect of self-made Sanjin-Weiwei decoction for patinets with upper urinary tract calculi after extracorporeal shock wave lithotripsy / 国际中医中药杂志

Objective To study the clinical effect of Self-made Sanjin-Weiwei decoction for patinets with upper urinary tract calculi after extracorporeal shock wave lithotripsy (ESWL). Methods A total of 200 patients with upper urinary tract calculi after ESWL in our hospital from January 2015 to Octomber 2016 were enrolled in this study. The subjects were divided into the control group (n=100) and the treatment group (n=100) randomly. The control group were treated with conventional treatment after ESWL, and the treatment group were treated with conventional treatment plus the Self-made Sanjin-Weiwei decoction after ESWL,and the two groups were treated for 45 days. The clinical effects of both groups were compared. The first time average row stone time and renal colic average frequency of both groups after treatment was compared. The stone platoon net rate of both groups after 1 month, 2 months, 3 months were compared. The incidence of adverse reactions, like dry mouth, nausea, painful urination, blood in urine, of both groups during the treatment were compared. Results The total effect rate of the treatment group was 86.0%, significantly higher than 62.0% of the control group(χ 2=14.969,P<0.001).The first time average row stone time(4.18 ± 1.30 d vs.7.52 ± 2.08 d,t=13.617)and renal colic average frequency(0.67 ±0.21 vs.1.55 ±0.87,t=9.833)of the treatment group were significantly lower than the control group (P<0.05). The stone platoon net rate of the treatment group after 1 month (93.0% vs.74.0%,χ 2=13.101),2 months(98.0% vs.82.0%,χ 2=14.222),3 months(100.0% vs.84.0%,χ 2=17.391)were significantly higher than the control group (P<0.05). The incidence of adverse reactions (7.0% vs. 15.0%, χ2=4.735)of the treatment group were significantly lower than the control group(P<0.05).Conclusions The Self-made Sanjin-Weiwei decoction for patinets with upper urinary tract calculi after ESWL has a good effect and low incidence of adverse reactions, can reduce the renal colic and improve the stone platoon net rate.

Preparation of sheddable PEG modified miRNA-complexing nanoparticles and its preliminary in vitro and in vivo study / 中国药科大学学报

@#This study was aimed to prepare sheddable PEG modified miRNA-complexing nanoparticles and investigate in vitro cellular uptake effect and in vivo distribution profile. The sheddable PEG material was synthesized through condensation. The sheddable PEG modified miRNA-complexing nanoparticles were successfully prepared by electrostatic interaction between gene vector and miRNA, and then ibuprofen was added to deshield PEG layer. The in vitro cellular uptake effect and in vivo distribution profile of nanoparticles were investigated on 4T1 model cells. As a result, the particle size of nanoparticles was 107. 7 nm and Zeta potential was 15. 8 mV. Compared to unsheddable PEG group, the cellular uptake effect by 4T1 tumor cells as well as the concentration on tumor regions was significantly improved in the sheddable PEG group. Results showed that this systen has a great potential application in the field of tumor treatment.

Role of vascular endothelial glycocalyx in inflammation / 国际生物医学工程杂志

Luminal surface of vascular endothelium is decorated with a variety of polysaccharide-protein complexes,which constitute the glycocalyx.It has been demonstrated that vascular endothelial glycocalyx plays an important role in modulation of selective permeability of vessels,mediation of the blood cell-endothelial cell interactions and the release of nitric oxide induced by fluid shear stress under physiological condition.In inflammation condition,sheding of glycocalyx due to inflammation mediator leads to its functional weakening in vessel protection.At the same time,heparan sulfate as a major constituent of vascular endothelial glycocalyx could be involved in regulating the evolution of inflammation.Heparan sulfate interacts with L-selectin to mediating leukocyte rolling,presents chemokines on luminal surfaces of endothelial cells to mediate leukocyte crawling and firm adhesion,participates in transcytosis of chemokines from tissue to luminal side of endothelial cells during inflammation.Various risk factors of atherosclerosis,as an inflammatory disease,are closely associated with vascular endothelial glycocalyx.This paper is aimed to review the role of vascular endothelial glycocalyx in inflammation and atherosclerosis.

Advance in preparation of magnetic nanowires and its application in biosensors / 国际生物医学工程杂志

Magnetic nanowires not only have nanometer properties, but also have magnetic property of giant magnetoresistance. Recently, nanowires were applied widely in high density hard disk slider, super magnetic storage element, micro-sensors, micro-engine, biomedical engineering, etc. This artical reviews the current preparation method of nanowires, the template synthesis, including several general templates such as track-etched porous polycarbonate membrane, porous anodic aluminum oxide membrane and carbon nanotubes, as well asgrowth patterns of nanowires in the templates. The applications of nanowires in several biosensors are introduced.The applications will greatly improve current ways of sensor detection, which will open up new areas of the field of biosensor study.

Numerical simulation of the distribution of shear stress on the bottom of parallel plate flow chamber under different inlet velocity conditions / 生物医学工程学杂志

The distribution of shear stress on the bottom of the parallel plate flow chamber under different inlet velocities was analyzed by numerical simulation. In the present experimental study, the projection planes of the relative errors at 0.7% level were obtained, and then the efficient region and the actual entrance length were further corrected by introducing the concept of relative error. The results showed that the efficient region of the chamber increased with the direction of length while the inlet velocity was increased, and the actual entrance length was much greater than that of the theoretical entrance length. Therefore, in accordance to the needed range of shear stress in experiment and to the needed efficient region area, the optimum design of the flow chamber is necessary.

Mechanical-chemical interaction and its mechanobiological mechanism on the migration of endothelial cells / 医用生物力学

Objective To elucidate the mechanical chemical interaction and its mechanobiological mechanism on the migration of endothelial cells. Method RT PCR, Western blot and immunofluorescence were applied to detect the expression of CXCR1 and CXCR2 and their distributions under three levels of shear stress; anti-IL8RA and anti-IL8RB were used to inhibit CXCR1 and CXCR2 to evaluate endothelial cell migration under shear stress; ECs were transfected to obtain the wild type Rac1(Rac1WT) or RhoA (RhoAWT), the constitutively active forms of Rac1(Rac1Q61L) or RhoA (RhoA63L), and the dominant negative forms of Rac1(Rac1T17N) or RhoA (RhoA188A) respectively, with lipofectamine 2000 reagent. ECs transfected with three plasmids of Rac1 were exposed to three levels of shear stress and IL-8, respectively; ECs transfected with three plasmids of RhoA were stimulated by IL-8. Results CXCR1 and CXCR2 are novel mechano sensors mediating laminar shear stress induced endothelial cell migration. High expression of Rac1 and RhoA can promote EC migration, while their low expression inhibits EC migration. Conclusions CXCR1, CXCR2, Rac1 and RhoA are critical signaling molecules in mechanical chemical interaction of EC migration.

The role of CXCR1/2 in shear stress-induced endothelial cell migration / 生物医学工程学杂志

CXCR1 and CXCR2 are important receptors in regulating vascular endothelial cell activities. In order to elucidate the role of CXCR1/2 in shear stress-induced endothelial cell migration, we have investigated the expression levels of CXCR1 and CXCR2 in the endothelial cells exposed to shear stress. In the experiment, anti-IL8RA and anti-IL8RB were used to antagonize CXCR1 and CXCR2. Different shear stresses were generated in a flow chamber; scratch test was carried out to compare endothelial cell migration in the control group and the receptor-antagonized groups. The results indicated that the migration of endothelial cells was restrained effectively after CXCR1 and CXCR2 were antagonized by anti-IL8RA and anti-IL8RB. And anti-IL8RA showed a stronger inhibitive effect than did anti-IL8RB (P<0.05). In the group with both receptor antagonisms, the migration was further inhibited. These results suggest that both CXCR1 and CXCR2 are important factors in mediating the migration of endothelial cells induced by shear stress, and CXCR1 fulfills a more important role.

Rac1 mediates the migration of endothelial cells induced by IL-8 / 生物医学工程学杂志

This study sought to elucidate whether Rac1 mediates the migration of endothelial cell induced by IL-8. The Transwell chamber motility assay was conducted to disclose the effect of different matrigel dilution and different time of IL-8 treatment on the migration of endothelial cells. The mRNA of Rac1 was detected by RT-PCR. The results demonstrated that when the concentration of Matrigel was 1:2, there is significant difference on the amounts of migration cells than that of the concentration of 1:3 or 1:8; When the dilution of Matrigel was 1:4, 1:5 or 1:6, there is no significant difference on the amounts of migration cells than that of other dilution groups. So we choose the Matrigel concentration as 1:4. With the increase of IL-8 stimulation time, the cells which migrated from upper reservoirs to lower reservoirs progressively increased. After six hours stimulation by IL-8, the expression of Rac1 mRNA in migrated cells was increased, compared with that of other groups. The results suggest that Rac1 may mediate the migration of endothelial cells induced by IL-8. It can also be the foundation for further investigation on the role of Rac1 in the migration of endothelial cells induced by IL-8.

Immune regulatory effect of natural killer T cells in the early stage of murine liver injury induced by type 5 adenovirus / 中华微生物学和免疫学杂志

Objective To study the immune regulatory effect of natural killer cell(NKT) in the early stage of murine liver injury induced by type 5 adenovirus(Ad5). Methods Animal models were con-structed by injected C57BL/6 mice with 1.5×109-3×109 PFU Ad5 into the tail vein. Liver injury of mouse at day 0, 1, 2, 3, 4, 5 after infection was determined by HE staining and serum ALT/AST(alanine amin-otransferase/aspartate aminotransferase) level. Flow cytometry analysis was used to measure the proportion of lymphocytes, expression of Fas/FasL on the surface of NKT cells and level of IL-4, IFN-γ/in NKT cell plasma in the infected mouse liver. RT-PCR was applied to semi-quantify the chemokines and their receptors mRNA in infected mouse liver. Results NKT cells of mouse increased significantly at day 1 after infected with high titer Ads(3×109 PFU), expression of FasL on NKT cell and plasma IL-4, IFN-γ/level in NKT cells were also up-regulated, hence the obviously infiltration of lymphocytes in routine liver. Comparing with high titer Ads infection, low titer Ads infection (1.5×109 PFU) lead to little change of NKT cell proper-tion, and fewer infiltration of lymphocytes in murine liver. Hepatic chemokine RANTES, 1P-10, and MIP-1β mRNA expression in C57BL/6 was up-regulated 2 d after intravenous administration of 3×109 PFU Ad5. Corresponding chemokine receptor CCR5, CCR1, CXCR3 mRNA expression was up-regulated 3 d after in-fection. Conclusion NKT cells play an important role in lymphocytes recruitment into the liver of mouse in-fected with AdS, which may relate to up-regulatio of the plasma IL-4, IFN-γ level and expression of FasL of NKT cells, therefore facilitating the production of chemokines, e.g. IP-10 and Mig.

Effects of laminar shear stress on the expression of IL-8 receptor CXCR1 in endothelial cells / 生物医学工程学杂志

This study was conducted to elucidate the effects of different fluid shear stress on the expression of IL-8 receptors CXCR1 in endothelial cells. The HUVEC cell lines, EA. Hy926 cells, were cultured in vitro, exposed to 5.56, 10.02, and 15.27 dyn/cm2 laminar shear stress, respectively, and then the shear stress grous were each investigated at the time-points of 0h, 1h, 2h, 4h and 8h. Western blotting was used for detecting the expression of IL-8 receptor CXCR1 at the time-points. The results showed that, under 5.56 dyn/cm2 shear stress, the expression of CXCR1 increased with time significantly (P < 0.01). The maximum expression of CXCR1 appeared at 4 h and was 2.2 times that of the control. When exposed to 10.02 dyn/cm2, the expression of CXCR1 increased gradually with time and finally remained at a constant higher level. When exposed to 15.27 dyn/cm2, CXCR1 expression decreased significantly with time (P < 0.01). The minimum CXCR1 expression appeared at 8 h and was 62. 59 percent of that of the control. These results indicate that the expression of CXCR1 in endothelial cell is regulated by laminar shear stress.

Holmium Laser Lithotripsy via Ureteroscope for Upper Urinary Tract Radiotransparent Calculi Complicated with Polyps / 中国微创外科杂志

Objective To investigate the effect,safety,and skills of holmium laser lithotripsy by ureteroscopy for upper urinary tract radiotransparent calculi complicated with polyps. Methods From January to December 2008,60 patients with upper urinary tract radiotransparent calculi complicated with polyps were treated by holmium laser lithotripsy(Coherent PowerSuite 20 W,500 ?m) via a ureteroscope.The laser lithotrite was introduced to the stones,and the intraluminal stone was fragmented and melted by holmium laser simultaneously.Results The mean operation time of this series was 21 min(15-70 min).The polyps was removed completely in one session in all of the cases except in 4 patients,whose upper calculi were flushed into the renal pelvis.No ureteral mucosa avulsion,perforation,false passage and other severe complications occurred.Some of the patients had mild haematuria after the operation without fever or renal pain.The patients were followed up for 3 to 12 months(mean 5 months);none of them had residual stones or ureteral stricture during the period. Conclusions Holmium laser lithotripsy via ureteroscope is a safe and effective procedure for the treatment of upper urinary tract radiotransparent complicated with polyps.

Recombinant adenovirus-mediated survivin gene transfection of dendritic cells induces immune responses against renal cell carcinoma / 中国肿瘤生物治疗杂志

Objective:To investigate the anti-tumor immune responses against renal cell carcinoma induced by dendritic cells(DCs)transfected with survivin gene mediated by recombinant adenovirus.Methods:The DCs derived from human peripheral blood were infected with recombined adenovirus vector carrying the survivin gene.The expression of survivin protein in the infected DCs was examined by Western blotting assay;the surface expression of CD83,MHCⅡ,CD80 and CD86 by flow cytometry(FCM),Interleukin-12(IL-12)in the supernatants of DCs and IFN-? released by the cytotoxic T lymphocytes(CTLs)by ELISA,the ability of DCs in proliferating allo-lymphocytes by mixed lymphocyte reaction(MLR),and specific killing activity of CTLs by MTT assay.Results:The DCs presented mature DCs phenotype after transfection with Ad-svv.The expression of survivin protein in transfected DCs was confirmed by Western blotting analysis.The IL-12 level in the supernatant of DCs transfected with Ad-svv was significantly higher than that transfected with empty vector(Ad-CMV,P

Distribution and Anti-microbial Resistance of Pathogenic Bacteria Isolated from Hospitalized Neonates in Wuhan Area / 中华医院感染学杂志

OBJECTIVE To analyze the distribution and anti-microbial resistance profile of pathogenic bacteria isolated from hospitalized neonates in Wuhan area.METHODS The strains of bacteria isolated from neonates in hospitalization of Wuhan area from 2006 to 2007 were identified by VITEK 32 automatic bacteria system and antimicrobial susceptibility test was performed by Kirby-Bauer method.The standards of NCCLS issued in 2006 were used to assess the results of antimicrobial susceptibility.RESULTS A total of 3892 strains of bacteria were isolated from 10053 specimens,the positive rate was 38.7%.The Gram-positive cocci and Gram-negative bacilli accounted for 41.0% and 55.2%,respectively.The leading 8 species on the list of isolated strains in the two years accounted for 90.8% of all the isolated strains.MRCNS accounted for 68.6% of coagulase negative staphylococcus,and the MRSA kept a low rate(2.6%) in Staphylococcus aureus.The infection rate of newborn by Enterococcus in blood was remarkably higher than that of the past report in this area.The ?-lactamases-producing Escherichia coli and Klebsiella pneumoniae were accounted for 46.9% and 34.7%,respectively.CONCLUSIONS The distribution and anti-microbial resistance of hospitalized neonates in Wuhan area have regional and group characteristics;it′s necessary to strengthen monitoring the regional epidemic characteristic status and the drug-resistant status of the pathogenic bacteria among neonates.