ABSTRACT
Biapenem is a carbapenem antibiotic， and can be used for the treatment of sepsis， pneumonia， lung abscess， chronic respiratory lesions secondary infection， complex urinary tract infection and pyelonephritis， etc. This article reviewed the studies on the pharmacokinetics， pharmacodynamics and therapeutic drug monitoring （TDM） of biapenem. The pharmacokinetic parameters of biapenem are not significantly different in healthy subjects， and there is no accumulation after multiple doses of biapenem. However， there are large differences in pharmacokinetic parameters in patients with severe disease and patients with abnormal renal function compared with healthy subjects， which leads to conventional treatment regimens not achieving the desired outcome. In terms of pharmacodynamics， biapenem can improve the rate of reaching the target value by increasing the frequency of administration and prolonging the infusion time. For patients with anuria in end-stage renal disease， dosing intervals can be extended to avoid drug accumulation. However, for patients with severe infection, a daily dose of 1.2 g still can not control infections caused by Acinetobacter baumannii or Pseudomonas aeruginosa, which limits its use in patients with severe disease. It is recommended to implement TDM in severe patients and patients with abnormal renal function， and explore the best dosing regimen for biapenem in combination with pharmacokinetic models to ensure that the time that the free blood concentration of biapenem remains above minimum inhibitory concentration as a percentage of the time between doses （%fT＞MIC） is within the effective range，so that biapenem can exert a greater efficacy in severe patients and patients with abnormal renal function. For medical institutions that cannot carry out TDM， the efficacy of biapenem can be maximized by increasing the frequency of administration and prolonging the infusion time. For infections caused by P. aeruginosa， A. baumannii and Serratia marcescens with high drug resistance rates， it is recommended to combine or replace other antibiotics.
ABSTRACT
In recent years， with the increasing understanding of the genetic mechanisms of hypertrophic cardiomyopathy， novel molecular-targeted drugs Mavacamten and Aficamten are two cardiac myosin inhibitors currently approved by the FDA for the treatment of hypertrophic obstructive cardiomyopathy. Both of them have a similar mechanism of action and can selectively bind to different variable sites of cardiac myosin to inhibit cardiac myosin， thus reducing myocardial hypercontractility. Relevant clinical studies have also shown that both drugs can reduce patients’ left ventricular outflow tract pressure gradient， the levels of N-terminal pro-B-type natriuretic peptide and cardiac troponin I as cardiac markers， and improve New York Heart Association （NYHA） cardiac function class. They are safe， have mild adverse reactions， and can be tolerated by patients. Compared to Mavacamten， Aficamten， as a structurally optimized product， has a shorter half-life and fewer drug-drug interactions， which is more conducive to drug- targeted dose titration.
ABSTRACT
Extracorporeal membrane oxygenation (ECMO), a kind of life support technology that can replace lung and heart function, is widely used in critical respiratory and circulatory exhaustion. Because of the serious diseases and the use of interventional catheters, patients receiving ECMO life support are often administrated with broad-spectrum antimicrobial agents, which increase the risk of fungal infection. Fungal infection during ECMO can increase mortality. How to effectively control fungal infection is a thorny problem faced by clinicians. During the treatment of ECMO, the patient's physiological status, ECMO oxygenation membrane, circulation pipeline and other factors may change the pharmacokinetic profiles of antifungal drugs, thereby affect the clinical efficacy of drugs. This artical reviews the pharmacokinetic characteristics of antifungal drugs during ECMO support, in order to provide references for clinical antifungal treatment.
ABSTRACT
Currently, HIV-1 vaccine development has still been a hot pot in the AIDS research. HIV-1 glycoprotein Env is the sole target in the virion surface that mediates the membrane fusion between the virion and cell in the HIV-1 infection process. Env protein is the significant immunogen for HIV-1 vaccine development. In recent years, there have been breakthroughs in the Env trimer research. For example, the strategies including SOSIP, NFL2P, and UFO had been applied to design and generate HIV-1 Env trimer. The improvement of quantity and stability is beneficial to achieve the HIV-1 native-like Env trimer for elicitation of strong neutralizing antibody responsing in animal immunization. This review focuses on the different strategies for Env trimer design and compares their advantages and disadvantages, combining with our work to give some advice, which might provide relevant information for the future HIV-1 immunogen design.
ABSTRACT
Objective To compare the clinical effects between laparoscopic radical resection combined with radiofrequency ablation (RFA) and open radical resection for colorectal liver metastases.Methods The retrospective cohort study was conducted.The clinicopathological data of 120 colorectal liver metastases patients who were admitted to the Mengchao Hepatobiliary Hospital of Fujian Medical University (80 patients) and the Third Mfiliated Hospital of Chongqing Medical University (40 patients) between September 2012 and April 2017 were collected.Sixty patients undergoing laparoscopic radical resection of colorectal cancer combined with RFA of liver metastases were allocated into the laparoscopy with RFA group,and 60 undergoing open radical resection of colorectal liver metastases were allocated into the open group.Observation indicators:(1) surgical and postoperative situations;(2) follow-up and survival situations.Follow-up using outpatient examination and telephone interview was performed to detect postoperative survival up to May 2017.Measurement data with normal distribution were represented as x±s,and comparisons between groups were analyzed using independent-sample t test.Measurement data with skewed distribution were described as M (range).Comparisons of count data were analyzed using chi-square test or Fisher exact probability.The repeated measures data were analyzed using the repeated measures ANOVA.The survival curve and rate were respectively drawn and calculated by the Kaplan-Meier method,and the Log-rank test was used for survival analysis.Results (1) Surgical and postoperative situations:① All the patients underwent successful surgery,without conversion to open surgery in the laparoscopy with RFA group.Operation time,volume of intraoperative blood loss,cases with overall complications,death,abdominal pain,nausea and vomiting,liver dysfunction,pleural effusion,non-calculus cholecystitis and peptic ulcer and duration of postoperative hospital stay were respectively (135±34)minutes,(451±197)mL,31,0,18,6,6,4,3,2,(13±4)days in the laparoscopy with RFA group and (165±49)minutes,(794±204)mL,42,1,15,9,10,11,5,5,(19±4)days in the open group,with statistically significant differences in operation time,volume of intraoperative blood loss,cases with overall complications and duration of postoperative hospital stay between groups (t =3.983,9.394,x2 =4.232,t =9.148,P＜0.05),and no statistically significant differences in cases with death,abdominal pain,nausea and vomiting,liver dysfunction,pleural effusion,non-calculus cholecystitis and peptic ulcer between groups x2 =0.376,0.686,1.154,3.733,0.134,0.607,P＞0.05).() Levels of alanine aminotransferase (ALT),total bilirubin (TBil) and prothronbin time (PT) before operation,at day 1 and 3 postoperatively were respectively (70±9)U/L,(399±36)U/L,(231±19) U/L,(21±3) μmol/L,(26±3) μmol/L,(23±5) μmol/L,(17.3±2.4) seconds,(20.2-±4.4) seconds,(18.9±2.8) seconds in the laparoscopy with RFA group and (68± 8) U/L,(412±39)U/L,(253±22)U/L,(21±4)μmol/L,(28±4)μmol/L,(27±8)μmol/L,(16.6±3.0)seconds,(22.1±5.2) seconds,(20.1± 4.4)seconds in the open group,with statistically significant differences in the levels of ALT,TBil and PT before operation,at day 1 and 3 postoperatively between groups (F=16.727,13.115,4.194,P＜0.05).(2) Follow-up and survival situations:120 patients were followed up for 7-24 months,with a median time of 20 months.The postoperative 1-and 2-year tumor-free survival rates,1-and 2-year overall survival rates were respectively 23.3％,11.9％,85.0％,40.0％ in the laparoscopy with RFA group and 20.0％,12.8％,83.3％,38.3％ in the open group,with no statistically significant difference in above indicators between groups (x2 =0.145,0.069,0.012,0.196,P＞0.05).Further analysis showed that postoperative 2-year overall survival rate of patients with 1,2,3 and ＞3 liver metastasis lesions were respectively 53.3％,38.2％,40.0％,16.7％ in the laparoscopy with RFA group and 50.0％,35.7％,40.0％,15.4％ in the open group,with a statistically significant difference in 2-year survival rate of patients with different liver metastasis lesions in the laparoscopy with RFA group (x2 =20.949,P＜0.05) and in the open group (x2 ＝ 21.349,P＜0.05).Conclusion There are some advantages of fewer traumas,less complications,faster postoperative recovery and minimally invasive in laparoscopic radical resection combined with RFA for colorectal liver metastases,meanwhile,less liver metastasis lesions and better prognosis are found.
ABSTRACT
Mycophenolate mofetil (MMF) is a cornerstone immunosuppressant in clinical therapy, such as organ transplantation. Because of the huge interindividual difference in genetic polymorphisms and some other, fixed-dose MMF-treated strategy can neither attain the targeted effective and safe MPA blood concentration nor satisfy the needs of individualized therapy,especially in children.The article reviews the value of genetic polymorphism of UGT,IMPDH,ABCC2,SLCO in MMF therapy in children to highlight its role in the precision treatment.
ABSTRACT
We constructed the CAP2NC prokaryotic expression vector of HIV-1 NL4-3 strain and obtained relatively pure CAP2NC protein by optimizing its purification conditions to explore its in vitro self-assembly conditions. Primers were designed according to the CAP2NC DNA sequence of HIV-1 NL4-3 strain. The target gene was amplified by PCR and cloned into prokaryotic expression vector pTO-T7. Then the recombinant strain was transformed into Escherichia coli BL21 (DE3). IPTG induced protein expression, then the protein was purified by hydrophobic chromatography. SDS-PAGE and Western blotting were performed to analyze the target protein, and the biological activity of the antigen was identified through ELISA. The self-assembly of CAP2NC protein was analyzed by transmission electron microscopy and gel filtration chromatography. The protein had good reaction with the specific antibodies of p24 and formed different structures in various conditions. When 10% yeast RNA was added to the protein complex, the recombinant protein only formed into a tubular structure, which was similar to the self-assembled structure of the HIV-1 virus capsid. The results showed that the HIV-1 CAP2NC protein had in vitro self-assembly activity, and the RNA affected the structure of CAP2NC protein assembly. The protein can be used as a simple and effective molecular model to study its structure, and then it can provide a reference for the study of HIV immature virus particles.
ABSTRACT
Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.
ABSTRACT
Objective To explore the effect of heme oxygenase-1 (HO-1) on expressions of hypoxia inducing factor 1 alpha (HIF-1α) and vascular endothelial growth factor (VEGF)and regeneration of hepatic vascular plexus after orthotopic liver transplantation ischemia-reperfusion injury in rats.Methods Theexperimental study was conducted.According to the random number table,240 SD rats were divided into the 3 groups,80 rats in each group.Empty virus group:rats were transfected with the empty virus.Induced group:rats were transfected with HO-1 overexpression adenovirus.Inhibited group:rats were transfected with HO-1 RNAi adenovirus.Rats were made pairs (1 ∶ 1) and established rat liver transplantation model according to two cuffs method.Rats with less weight and with heavier weight were respectively chosen as donor rats and recipient rats,and then recieved tail intravenous injection of adenovirus at 24 hours before operation.(1) Detection of transfection efficiency of adenovirus before operation:HO-1 expression of liver tissue of rats in each group was detected by Western blot at 12 and 24 hours after injection of adenovirus.(2) Liver function test of recipient rats after liver transplantation:liver functions of recipient rats [alanine transaminase (ALT),aspartate transaminase (AST),alkaline phosphatase (ALP),gamma-glutamyl transferase (GGT)] were detected at l,3,7 and 14 days postoperatively.(3) Pathological histology of liver tissue and injury scores of recipient rats in the 3 groups after liver transplantation:paraffin sections of recipient rats in the 3 groups at postoperative 1 and 14 days were stained by HE staining and observed by light microscope,and were evaluated by Suzuki damage score standard.(4) Relative expressions of HIF-1α,VEGF and HO-1 in liver tissue of recipient rats were detected by Western blot.(5) Von Willebrand factor (vWF) in liver tissue of recipient rats at 14 days postoperatively was detected by immunofluorescence staining and small vessels were counted.Measurement data with normal distribution were represented as x ±s.Comparison between groups was analyzed by the independent-sample t test,comparison among groups was done using one-way ANOVA,and pairwise comparison was analyzed by the LSD test.Results (1) Detection of transfection efficiency of adenovirus before operation:the relative expression of HO-1 of liver tissue of rats at 12 and 24 hours preoperatively after injection of adenovirus was 1.08±0.16 and 1.08±0.26 in the empty virus group,1.18±0.21 and 1.39±0.19 in the induced group,0.87±0.26 and 0.57±0.12 in the inhibited group,respectively,with statistically significant differences in different time points (F =4.232,36.513,P＜ 0.05).(2) Liver function test of recipient rats after liver transplantation:level of ALT at 3 days postoperatively in the empty virus group,induced group and inhibited group was (504±67)U/L,(438±47)U/L and (490±39)U/L,with a statistically significant difference (F=3.517,P＜0.05).Levels of ALT,AST and ALP at 7 days posto-peratively were (443±49) U/L,(430± 34) U/L,(455± 38) U/L in the empty virus group and (382± 49) U/L,(372±50) U/L,(394±25) U/L in the induced group and (493±44) U/L,(455±62) U/L,(470±72) U/L in the inhibited group,respectively,with statistically significant differences (F =10.950,5.667,5.398,P＜0.05).Levels of ALT,AST,ALP and GGT at 14 days postoperatively were (394±46)U/L,(361 ±68)U/L,(417 ±17)U/L,(4.5±1.1)U/L in the empty virus group and (283±47) U/L,(288±60) U/L,(332±46) U/L,(2.5±0.5) U/L in the induced group and (446± 43) U/L,(422± 51) U/L,(423± 63) U/L,(4.3 ± 1.3) U/L in the inhibited group,respectively,with statistically significant differences (F=26.906,9.924,8.013,9.279,P＜ 0.05).(3) Pathological histology of liver tissue and injury scores of recipient rats in the 3 groups after liver transplantation:liver cell swelling,loose cytoplasm and a varying quantity of inflammatory cell infiltration in the portal regions in the liver tissue of 3 groups were observed at 1 day postoperatively.A few inflammatory cell infiltrations in the portal regions,basically normal liver cell arrangement and a slightly swelling of liver cell were found in the empty virus group at 14 days postoperatively.Reduced liver cell swelling and basically normal structure of liver lobule were observed in the induced group.There were small patchy or focal necrosis of liver cell,masses of inflammatory cell infiltration in the portal regions and damage of bile duct in the inhibited group.Suzuki score at 1 day postoperatively in the empty virus group,induced group and inhibited group were respectively 6.7± 1.7,6.1 ± 1.2 and 7.6± 1.3,with no statistically significant difference (F=2.257,P＞0.05).Suzuki score at 14day postoperatively in the empty virus group,induced group and inhibited group were respectively 4.0±0.8,2.9± 0.8 and 5.1± 1.4,with a statistically significant difference (F=9.776,P＜0.05).(4) Western blot results:the relative expressions of HIF-1α and VEGF (43 KD) in liver tissue of recipient rats at 1 day postoperatively were 0.21±0.10,0.30±0.12 in the empty virus group and 0.23±0.09,0.34±0.14 in the induced group and 0.17± 0.06,0.29±0.11 in the inhibited group,respectively,with no statistically significant difference (F =0.902,0.410,P＞0.05).The relative expressions of VEGF (24 KD) and HO-1 in liver tissue of recipient rats at 1 day postoperatively were 1.21 ±0.25,0.55±0.12 in the empty virus group and 2.13±0.40,0.72±0.12 in the induced group and 0.91±0.22,0.26±0.07 in the inhibited group,respectively,with statistically significant differences (F=35.158,39.082,P ＜ 0.05).The relative expressions of HIF-1α,VEGF (43 KD),VEGF (24 KD) and HO-1 in liver tissue of recipient rats at 7 days postoperatively were 0.49±0.22,0.46±0.13,0.98± 0.37,0.98±0.37 in the empty virus group and 0.83±0.26,0.63±0.19,1.60±0.33,1.49±0.46 in the induced group and 0.24±0.09,0.30±0.12,0.64±0.18,0.75±0.26 in the inhibited group,respectively,with statistically significant differences (F=16.853,10.021,20.756,8.156,P＜0.05).(5) Immunofluorescence staining results:number of small vessels at 14 days postoperatively in the empty virus group,induced group and inhibited group was respectively 7.9±2.0,10.6± 1.9 and 7.6 ± 1.9,with a statistically significant difference (F=5.921,P＜0.05).Conclusion HO-1 could promote expressions of HIF-1α and VEGF in liver tissue after liver transplantation ischemia-reperfusion injury and regeneration of intrahepatic vascular plexus,and it also alleviate bile duct ischemia-reperfusion injury after liver transplantation.
ABSTRACT
Objective@#To study the clinical effect of high pressure oxygen and Butylphthalide in the recovery of cerebral metabolism after carbon monoxide poisoning.@*Methods@#84 patients treated from May 2014 to May 2016 in our hospital were selected. The subjects were randomly and equally divided into two groups. The control group adopted the conventional therapy and high pressure oxygen; on the basis, the observation group also took Butylphthalide. The clinical effect, duration of coma, recovery of consciousness, incidence rate of delayed encephalopathy was observed. After 1m of treatment, the HDS point was evaluated.@*Results@#The total effective rate of control group (76.19%, 32/42) was lower than that of observation group (95.24%, 40/42) (P<0.05) . The duration of coma for observation group was shorter than that of control group. The percentage for patients with recovery of consciousness and incidence rate of delayed encephalopathy for observation group was better than that of control group (P<0.05) . The HDS point for observation group was even higher than that of control group (P<0.05) .@*Conclusion@#The high pressure oxygen and butylphthalide can improve the clinical effective rate, shorten the duration of coma and promote the patient’s recovery of consciousness. It is worthy of clinical promotion.
ABSTRACT
Objective To establish an efficient baculovirus-insect cell expression system for the production of human immunodeficiency virus-1 ( HIV-1 ) envelope glycoprotein gp120 and to evaluate the physiochemical properties, antigenicity and immunogenicity of the recombinant protein. Methods The gene encoding HIV-1 NL4-3 gp120 was cloned into the downstream of pH promoter of the baculovirus transfer vec-tor pAcgp67B to construct the recombinant transfer vector pAc-gp120. Expression of the protein of interest was induced in baculovirus-infected High FiveTM insect cells. ELISA, analytical ultracentrifugation and size-exclusion chromatography were carried out to characterize physicochemical properties of the expressed gp120 protein. Immunogenicity of the recombinant gp120 protein was analyzed by HIV neutralization assay after im-munizing BALB/c mice with it. Results The recombinant HIV-1 gp120 protein was successfully obtained from the established insect cell expression system with a purity of more than 90％ and a mean yield of 13 mg/L in four batches. That recombinant HIV-1 gp120 protein was characterized by homogeneity in solution and possessed a good immunoreactivity to neutralizing antibodies and antisera against HIV. Immunogenicity analysis in BALB/c mice demonstrated that the recombinant gp120 protein could induce effective immune re-sponses against HIV-1 NL4-3. Conclusion A simple and scalable approach to obtain homogeneous and im-munogenic HIV-1 gp120 antigen is successfully established, which will promote further investigation of HIV vaccine candidates.
ABSTRACT
Objective To investigate the use of salvianolate injection for inpatients in our hospital; To primarily evaluate its rationality and security in clinic.Methods Medical records using the salvianolate injection from December 2013 to March 2014 were surveyed statistically and analyzed in respect of patient gender and age, department, disease diagnosis, usage and dosage, solvent types, treatment course, medicine combination, occurence of ADR, etc.Results Totally 600 cases of inpatients receiving salvianolate injection were collected, about 69.33% consistent with the indication, 67.00% with a correct single dosage, and 52.50% with a correct treatment course. In the clinical application of salvianolate injection, a high rate of unreasonable usage was found in indication, single dosage, treatment course and medicine combination.ConclusionBecause the problems still existed in the use of salvianolate injection, it should be used strictly according to the medicine instructions and General Principles for Clinical Application of Traditional Chinese Medicine Injection to ensure clinical medication safety.
ABSTRACT
@#A long chain structure of DNA(polyaptamer)composed of multiple aptamer units was synthesized by rolling circle amplification and used for the construction of polyaptamer-doxorubicin system in the treatment of leukemia cells. It was found that the system was significantly more effective than monoaptamer in targeting and killing leukemia cells as it provided 35-fold enhanced binding affinity and 10-fold greater drug loading via multivalent effects. Drug release and cell viability also proved that the conjugates could gain entrance into the cells and rapidly release doxorubicin under the action of lysosome, leading to the tumoricidal effect.
ABSTRACT
Objective To investigate and analyze the military operation ability and influence factors of soldiers alongthe border in an extremely cold environment and to provide a scientific basis for effectively improving the combat effective -ness of the army stationed the cold regions.Methods According to relevant standards, recruits(the length of service 1 year) were randomly selected to investigate their military operationfactors, physical condition, sleep condition, mental health and cognitive ability.Results Cold was the main factor whichaffected military operation capability in cold regions.According to physical standards,border troops were in poor physicalcondition.Results of SCL-90 showed that the total score, the number of positive items , the score of each factor of the 130recruits were significantly lower than those of the Chinese army model and 46 veterans (P <0.05).There was nosignificant difference in these aspects between the 46 veterans and Chinese army model.Recruits were in better sleepcondition and had better instantaneous memory(P <0.05).Birthplace had no effect on these factors .Recruits from Centraland South China were in poorer sleep condition than those from cold regions (P <0.05).Results of SCL-90 showed that thetotal score,the factors of somatization, coercion, depression and anxiety of communication veterans were significantly higherthan those of the Chinese army model (P <0.05).The sleep condition of communication veterans was poorer than that ofmotorized infantry and patrol veterans (P <0.05).Conclusion Cold is the main factorthat impacts the ability of militaryoperations in cold regions.The physical work capacity of border guards in cold regions was significantly below the militarystandard, so the level of military training should be strengthened .Research on cold-protection equipment for special tasks should be strengthened.
ABSTRACT
Objective To investigate the effect of plateau training on military operation ability ,evaluate the real situa-tion of combat forces on the plateau , and to provide data for effectively improving the combat effectiveness of the army stationed on the plateau .Methods According to the standards , the levels of high altitude acclimatization , physical work capacity (VO2max, PWC170) and mental work capacity (digit span, digit symbol, pursuit aiming and visual reaction time) were measured to assess the military operation ability of plateau training troops , including troops that entered the plateau from the plain(plateau-entering troops), troops stationed on the plateau (plateau troops) and troops stationed in the plain ( plain troops ) .Results The troops on the plateau all reached basic acclimatization .The aerobic capacities of these three types of troops were of the standard medium level , and the maximum oxygen uptake ( VO2max ) of the plain troops was greater than that of that of plateau-entering troops(P<0.01), and the VO2max of the plateau troops was greater than that of plateau-entering troops(P<0.01).Compared with the plain troops, the VO2 max and PWC170 of the plateau-entering troops decreased by 25.7%and 27.7%respectively.There was no significant difference in the digit span , digit symbol, pursuit aiming between these 3 kinds of troops , but the visual response of the plateau-entering troops was prolonged ( P<0.01), while the pursuit aiming of the plateau troops was lower than that of the plateau-entering troops(P <0.05). Conclusion Acclimatization-promoting measures and plateau training can contribute to high altitude acclimatization formation, but cannot overcome the objective adverse effects of the plateau on physical fitness .Two months of plateau training fails to improve physical fitness.Therefore, special attention should be paid to the research on scientific and effective measures to improve physical fitness on the plateau .
ABSTRACT
Induced pluripotent stem cells (iPSCs) are derived from differentiated somatic cells through reprogramming in vitro.Similarly to embryonic stem cells (ESCs),iPSCs have the ability of unlimited selfrenewal and multiple differentiation potential.But they are without the constraints of immunology and ethics and acquire conveniently.Patient-specific iPSCs provide an invaluable resource for disease modeling,drug discovery,regenerative therapy,immune therapy and cellular delivery vehicle in cancer therapy.Nowadays,the application of iPSCs technology has made some progress in the treatment of tumors.
ABSTRACT
Matrix protein 2(M2) is an integral tetrameric membrane protein of influenza A virus, which functions as ion channel. M2 sequence has shown remarkable conservation, so there has been growing interest in it as "universal" vaccine. In order to establish a stable 293 cell line that express M2 protein under the control of the tetracycline operator, M2 gene was obtained by PCR amplification from the plasmid containing the segment 7 of influenza A virus strain A/PR/8/34 firstly. The PCR product was cloned into BamH I/Not I restriction site of pcDNA5/FRT/TO vector, and cotransfected with pOG44 which express Flp recombinase into Flp-In T-REx-293 cell. Integration of pcDNA5/FRT/TO-M2 into the cell genome at the Flp Recombination Target (FRT) site brought the SV40 promoter and the initiation codon in frame with the hygromycin resistance gene. Thus, stable cell lines were selected for hygromycin resistance. The expression of M2 protein from hygromycin-resistant cell was induced by addition of tetracycline into the cell culture media, and then tested by indirect immunofluorescence assay (IFA). 16 strains with high expression of M2 were selected. After subculturing for more than ten passages, the cell lines still stably expressed M2 protein. No M2 protein could be detected without tetracycline induction, suggesting that the expression was strictly controlled by tetracycline operator. The cell lines expressing M2 will be useful for further functional studies of M2 protein, detection of immune response against natural structure M2 protein and development of live attenuated influenza virus vaccine with reverse genetics technique.
Subject(s)
Animals , Humans , Cloning, Molecular , Gene Expression , Genetic Vectors , Genetics , HEK293 Cells , Influenza A virus , Genetics , Metabolism , Influenza Vaccines , Genetics , Operator Regions, Genetic , Recombinant Proteins , Genetics , Tetracycline , Pharmacology , Transfection , Viral Matrix Proteins , GeneticsABSTRACT
In order to evaluate the response to vector-expressed M1 and HA genes of influenza virus in mice, we prepared recombinant plasmid pStar-M1/HA and recombinant adenovirus Ad-M1/HA containing both the full-length matrix protein 1(M1) and hemagglutinin (HA) genes of human H5N1 influenza virus strain A/Anhui/1/2005. We then combined the DNA vaccine and adenoviral vaccine in immunization of BALB/c mice with a prime-boost regime. We immunized the mice with DNA vaccine at day 0 and 28 and with recombinant adenoviral vaccines at day 14 and 42. We took blood samples before each injection and 14 days after the final injection for detection of humoral immune responses. At day 56, we sacrificed the mice and collected splenocytes for detection of cellular immune responses. ELISA and hemagglutination inhibition (HI) assay showed that specific IgG Abs against H5N1 influenza virus was induced in serum of the immunized mice. ELISPOT results confirmed that the specific cellular immune responses were successfully induced against the M1 and HA proteins of H5N1 influenza virus. This study provides new strategy for development of novel influenza vaccines.
Subject(s)
Animals , Mice , Adenoviridae , Genetics , Metabolism , Antibodies, Viral , Blood , Hemagglutinin Glycoproteins, Influenza Virus , Genetics , Allergy and Immunology , Immunization , Influenza A Virus, H5N1 Subtype , Allergy and Immunology , Influenza Vaccines , Allergy and Immunology , Mice, Inbred BALB C , Recombinant Fusion Proteins , Genetics , Allergy and Immunology , Vaccines, DNA , Allergy and Immunology , Viral Matrix Proteins , Genetics , Allergy and ImmunologyABSTRACT
We developed vectors expressing two antigen of H5N1 influenza virus. Based on the human H5N1 avian influenza virus strain A/Anhui/1/2005 isolated in China, we amplified the matrix protein 2 (M2) and Hemagglutinin (HA) genes by PCR and subcloned them into pStar vector to construct two genes co-expressing recombinant DNA vaccine pStar-M2/HA. After transfection of 293 cells with the plasmid, we confirmed with indirect immunofluorescence assay (IFA) that M2 and HA genes cloned on plasmid pStar co-expressed successfully. Using Ad-Easy adenovirus vector system, by homologous recombination in bacteria and packaging in 293 cells, we constructed two recombinant adenoviruses, namely Ad-M2 and Ad-HA. After infection of 293 cells with the recombinant adenoviruses, we confirmed with IFA that M2 and HA genes cloned into adenoviruses expressed successfully. We then combined the recombinant DNA vaccine and adenoviral vector vaccines in immunization of BALB/c mice with a prime-boost regime. On day 0 and day 28, we immunized the mice with DNA vaccine and on day 14 and day 42, with recombinant adenovirus vaccines. We took blood samples before each injection and 14 days after the final injection. On day 56, we collected splenocytes from the mice. ELISA and hemagglutination inhibition (HI) assay showed that the vaccines successfully induced specific IgG antibodies against HA protein in serum of the immunized mice. ELISPOT confirmed that the vaccines successfully induced the special cellular immune response to M2 and HA protein of H5N1 influenza virus. The study on combined immunization with M2 and HA genes provided basis for development of novel influenza vaccine.
Subject(s)
Animals , Female , Mice , Adenoviridae , Genetics , Metabolism , Genetic Vectors , Genetics , Hemagglutinin Glycoproteins, Influenza Virus , Genetics , Influenza A Virus, H5N1 Subtype , Genetics , Allergy and Immunology , Influenza Vaccines , Allergy and Immunology , Mice, Inbred BALB C , Recombinant Proteins , Genetics , Allergy and Immunology , Vaccination , Vaccines, DNA , Allergy and Immunology , Viral Matrix Proteins , GeneticsABSTRACT
Changes of functional connectivity network of human V5 in different brain activity was investigated by combining spatial independent component analysis with temporal correlation. First, V5 was localized by performing spatial independent component analysis on the data from block design visual motion runs; then low frequency correlations between V5 and other regions were computed in two steady states (resting state and the state with continuous visual motion stimulus) to detect the functional connectivity networks. The results of experiment indicated: The functional connectivity network of V5 was more extensive and was consistent with the known anatomical connectivity during rest; when subjects were viewing motion, the network was limited in the visual cortex, suggesting that V5 was acting in concert with a network specific to the visual motion processing task.