Evolution and development of potent monobactam sulfonate candidate IMBZ18g as a dual inhibitor against MDR Gram-negative bacteria producing ESBLs

A series of new monobactam sulfonates is continuously synthesized and evaluated for their antimicrobial efficacies against Gram-negative bacteria. Compound 33a (IMBZ18G) is highly effective in vitro and in vivo against clinically intractable multi-drug-resistant (MDR) Gram-negative strains, with a highly druglike nature. The checkerboard assay reveals its significant synergistic effect with β-lactamase inhibitor avibactam, and the MIC values against MDR enterobacteria were reduced up to 4-512 folds. X-ray co-crystal and chemoproteomic assays indicate that the anti-MDR bacteria effect of 33a results from the dual inhibition of the common PBP3 and some class A and C β-lactamases. Accordingly, preclinical studies of 33a alone and 33a‒avibactam combination as potential innovative candidates are actively going on, in the treatment of β-lactamase-producing MDR Gram-negative bacterial infections.

Clinical study of acupuncture combined with medication for the elderly with Alzheimer disease / 针灸推拿医学（英文版）

Objective: To observe the impact of mind-regulating acupuncture plus donepezil on the cognitive ability, mean cerebral blood flow velocity, event-related potential P300, and activities of daily living (ADL) in the aged patients with Alzheimer disease (AD).Methods: Sixty senile AD patients were divided into a treatment group and a control group following the envelope method for random allocation, with 30 cases in each group. Based on the conventional treatment of the internal medicine, the control group received oral donepezil, and the treatment group received oral donepezil plus mind-regulating acupuncture. After 4-week treatment, the two groups were evaluated by the mini-mental state examination (MMSE), Alzheimer disease assessment scale-cognitive part (ADAS-Cog), and ADL; changes in P300 and the mean cerebral blood flow velocity were also observed.Results: Before treatment, there were no significant differences in the scores of MMSE, ADAS-Cog, or ADL between the two groups (P>0.05). The MMSE score increased after treatment in both groups and was notably higher in the treatment group than in the control group, showing intra-group and inter-group statistical significance (P<0.05). After treatment, the ADAS-Cog and ADL scores dropped in both groups and were markedly lower in the treatment group than in the control group, also showing intra-group and inter-group statistical significance (P<0.05). Compared with the same group before treatment, the latency of P300 was shortened and the amplitude was extended in both groups, all with statistical significance (P<0.05); the latency was shorter and the amplitude was larger in the treatment group than in the control group after treatment, presenting significant between-group differences (P<0.05). The mean blood flow velocity accelerated after the intervention in both groups, and the differences were statistically significant (P<0.05); the improvement in the treatment group was more notable than that in the control group (P<0.05).Conclusion: Mind-regulating acupuncture plus donepezil can regulate the latency and amplitude of P300, increase cerebral blood flow, and improve the learning and memory abilities of AD patients.

The exploration of the significance of proactive healthy living index for the elderly based on the functional capacity of Chinese older people / 中华老年医学杂志

Objective:To explore the significance of proactive healthy living index(PHLI)to provide a reference for the elderly to maintain function through healthy daily life.Methods:The functional status of the elderly was defined based on gait speed and cognitive function.Logistic regression model was used to select the daily life indicators which could be the risk factors for the functional status, and a nomogram was graphed for its visualization.PHLI was formed according to the result of the nomogram.Receiver operating characteristic curve(ROC)was plotted according to the functional status of the elderly, and the cut-off value of PHLI was determined based on the principle of the maximum sum of sensitivity and specificity.Results:A total of 4 971 participants with an average age of (70.9±7.5)years were included in the cross-sectional study, of whom 2 649 cases(53.3%)maintained normal function.The Logistic regression analysis showed that the housework had the most significant effect on the functional status of the elderly, followed by positive psychology( OR=2.04, 1.92; both P<0.001), increasing the likelihood of maintaining normal function by 104% and 92%, respectively.Regular life patterns, adequate protein intake, sleep quality and so on also influenced functional status of the elderly(all P<0.05). The PHLI median score of the elderly was 7.3(5.9, 8.3)points.The area under the curve(AUC)was 0.70(95% CI: 0.68-0.71), and the cut off value of PHLI was 6.8 points. Conclusions:The PHLI is associated with the maintenance of functional capacity in the elderly, which can be applied as a simple tool to provide reference for old people to keep functional health through living a proactive healthy daily life.

Feasibility, efficacy and safety of transbrachial access for interventional therapy on paravalvular leak post surgical valve replacement / 中华心血管病杂志

Objective: To investigate the feasibility, efficacy and safety of transbrachial access for interventional therapy on prosthetic paravalvular leak (PVL) post surgical valve replacement. Methods: This is a retrospective study. Patients with PVL after surgical valve replacement who underwent interventional therapy via the brachial artery approach in Structural heart disease center of Fuwai hospital between August 2017 and October 2019, were included. All patients underwent puncture of the brachial artery under local anesthesia, angiography and transcatheter closure procedure were performed. The procedure was performed under transthoracic echocardiography (TTE) guidance. Baseline data, operation data and pre-and post-operative TTE examination results were collected and analyzed. Postoperative complications were recorded and operational adverse events were obtained during follow up in the outpatient department after discharge. The operation success rate was calculated, which was defined as the degree of perivalvular regurgitation decrease by 1 grade and above according to TTE without interfering the valve movement and coronary artery blood flow within 30 days after occluder placement. Results: A total of 10 patients were enrolled in this study, the mean age was (57.5±14.6) years, and 6 patients were males. There were 7 cases with aortic PVL, and 3 cases with mitral PVL. Except for one patient who was converted to the femoral vein-transseptal approach, the other 9 patients were successfully implanted with the devices via the brachial artery approach. The operation time was (103.3±34.0) minutes, and there was no need for rigorous bed rest after the operation. The median hospital stay was 7.5 (3.0, 9.8) days. The operation success rate was 9/10 via the brachial artery approach. The differences in the degree of perivalvular regurgitation, New York Heart Association (NYHA) classification, left ventricular end diastolic diameter and left atrial diameter before and after operation were statistically significant (all P<0.05). One case developed new hemolysis with renal insufficiency on the second day after procedure and discharged after successful dialysis. Another case experienced complication of brachial artery pseudoaneurysm after procedure and discharged after successful treatment with thrombin injection. The mean follow-up time was (14.3±7.9) months. During the follow-up, NYHA classification remained as Ⅰ/Ⅱ in 9 patients, no operational adverse events were observed. Conclusions: Transbrachial access for interventional therapy on PVL post surgical valve replacement is a feasible, effective, and safe procedure. It has the advantages of simplifying the operation process and reducing postoperative bed rest time.

Effect of electroacupuncture at governor vessel on learning-memory ability and serum level of APP, Aβ in patients with Alzheimer's disease / 中国针灸

OBJECTIVE@#To compare the therapeutic effect of electroacupuncture (EA) combined with donepezil hydrochloride and donepezil hydrochloride alone on improving learning-memory ability in patients with Alzheimer's disease (AD), and to explore its action mechanism.@*METHODS@#Sixty patients of AD were randomly divided into an observation group and a control group, 30 cases in each group. The patients in the observation group were treated with EA at governor vessel (GV) combined with donepezil hydrochloride. EA was applied at Baihui (GV 20) and Fengfu (GV 16) with dilatational wave (10 Hz/50 Hz of frequency, 0.5 to 5.0 mA of intensity), and the needles were kept for 40 min, EA was given once a day; the donepezil hydrochloride tablet was taken orally, 5 mg, once a day, and after 4 weeks the dosage might be increased to 10 mg per day according to the specific situation. All the treatment was given for 8 weeks. The patients in the control group were only treated with donepezil hydrochloride with the identical procedure as the observation group. The Montreal cognitive assessment (MoCA) and Alzheimer's disease assessment scale cognitive part (ADAS-Cog) were evaluated before and after treatment; P300 (latency and amplitude of N2 and P3) was detected by EEG/ERP system brain event related potential instrument, and amyloid precursor protein (APP) and β-amyloid protein 1-42 (Aβ) were detected by ELISA.@*RESULTS@#Compared before treatment, the MoCA scores were increased after treatment in the two groups (<0.05), and the MoCA score in the observation group was higher than that in the control group (<0.05). Compared before treatment, the ADAS-Cog scores were decreased after treatment in the two groups (<0.05), and the ADAS-Cog score in the observation group was lower than that in the control group (<0.05). Compared before treatment, the latency of N2 and P3 was shortened and the amplitude was increased after treatment in the two groups (<0.05); after treatment, the latency of N2 and P3 in the observation group was shorter than that in the control group and the amplitude was higher than that in the control group (<0.05). Compared before treatment, the serum levels of APP and Aβ were lower after treatment in the two groups (<0.05), and the serum levels of APP and Aβ in the observation group were lower than those in the control group (<0.05).@*CONCLUSION@#EA at Baihui (GV 20) and Fengfu (GV 6) combined with donepezil hydrochloride can effectively reduce the serum levels of APP and Aβ and improve the scores of MoCA and ADAS-Cog and the levels of N2 and P3 of P300 in AD patients, which has superior effect to donepezil hydrochloride alone in improving the learning-memory of AD patients.

Goal management plays an important role in improving the scientific and technological strength of research-oriented hospitals / 中华医学科研管理杂志

Objective:To explore how to enhance the strength of science and technology of research-oriented hospital using goal management, thereby effectively promoting the construction and development of research hospitals.Methods:This paper studies the goal management measures and their effectiveness through data analysis, analyzes the problems of scientific research, goal management measures and changes in scientific research before and after the implementation measures over the years.Results:The goal management significantly enhances the scientific and technological strength. Total amount of research funding kept increasing year by year. Goal accountability management effectively improve the participation in individual scientific research activities. The implementation of high-quality papers goal management has a significant role in promoting the researchers to publish their achievements in domestic journals with international influence.Conclusions:Goal management plays an important role in the improvement of the scientific and technological strength of the hospital, which can effectively promote the construction and development of the research-oriented hospitals connotation. Goal management can serve as a powerful grasper for the research-oriented hospital to improve the scientific research capacity.

The relationship between doing housework and gait speed among the elderly / 中华老年医学杂志

Objective:To explore the relationship between doing housework and gait speed(GS)in order to identify ways to maintain physical capability in elderly people.Methods:A nationwide cross-sectional survey was conducted from Jan.2011 to Dec.2011.Subjects in this survey, with a sample of 2 519 males and 2 541 females aged ≥60 years, came from six regions across the country and satisfactorily completed a GS test.A GS in the lower 20% of the reference population(<0.7 m/s)was used as the cut-off point to define slow GS(SGS). Housework was defined as activities such as house cleaning, shopping, cooking and taking care of grandchildren and a response of"often"(≥5 d/week)was considered as doing housework.Propensity Score Matching(PSM)was performed to select comparable sub-groups of the study population to evaluate the association between housework and GS.Results:There were 3 290 subjects(1 645 pairs)selected by PSM.Logistic regression analysis showed that doing housework was inversely associated with SGS, and housework activities reduced the risk of SGS by 60%( OR=0.40, 95% CI: 0.35-0.46, P<0.01). Conclusions:Elderly people who conduct housework often are more likely to maintain GS and therefore should be encouraged to engage in daily activities to help maintain adequate physical performance.

Evaluation of GeneXpert MTB/RIF and BACTEC-MGIT 960 for the detection of tuberculosis among pneumoconiosis-associated tuberculosis patients / 中华劳动卫生职业病杂志

Objective@#To evaluate the performence of GeneXpert MTB/RIF and BACTEC-MGIT 960 on detecting Mycobacterium tuberculosis and rifampicin resistance for pneumoconiosis-associated tuberculosis patients.@*Methods@#The recruited 133 suspected active pneumoconiosis-associated tuberculosis hospitalized cases, morning sputum samples were collected to do modified L-J culture, conventional proportion method drug susceptibility test, GeneXpert MTB/RIF and BACTEC-MGIT 960. Analyze the sensitivity and specificity of the 133 sputum from patients, the positive rates of patients with tuberculosis in GeneXpert MTB/RIF test, BACTEC-MGIT 960 and modified L-J culture were 37.59%, 34.59% and 30.08% respectively. There was no significant difference among the three tests respectively (P>0.05) . According to the modified L-J culture, the sensitivity of GeneXpert MTB/RIF and BACTEC-MGIT 960 in detecting tuberculosis were 92.5% and 95.0% respectively, and specificity in rifampicin resistance were 86.0% and 91.4% respectively. There was no significant difference between GeneXpert MTB/RIF and BACTEC-MGIT 960 (P>0.05) . According to conventional proportion method drug susceptibility test, the sensitivity of GeneXpert MTB/RIF and BACTEC-MGIT 960 in detecting rifampicin resistance were 90.0% and 100%, and specificity were 92.6% and 96.4%. There was no significant difference between GeneXpert MTB/RIF and BACTEC-MGIT 960 (P>0.05) .@*Conclusion@#The GeneXpert MTB/RIF has good performence of detecting tuberculosis and rifampicin resistance. It has good application value among pneumoconiosis-associated tuberculosis patients.

and activity of d-serine in combination with -lactam antibiotics against methicillin-resistant

As d-amino acids play important roles in the physiological metabolism of bacteria, combination of d-amino acids with antibiotics may provide synergistic antibacterial activity. The aim of the study was to evaluate and activity of d-serine alone and in combination with -lactams against methicillin-resistant (MRSA) strains, and to explore the possible sensitization mechanisms. The activity of d-serine, -lactams alone and in combinations was evaluated both by standard MICs, time-kill curves and checkerboard assays, and by murine systemic infection model as well as neutropenic thigh infection model. An synergistic effect was demonstrated with the combination of d-serine and -lactams against MRSA standard and clinical strains. Importantly, the combinations enhanced the therapeutic efficacy in the animal models as compared to -lactam alone groups. Initial mechanism study suggested possible revision of d-alanine-d-alanine residue to d-alanine-d-serine in peptidoglycan by adding of d-alanine in the medium, which may cause decreased affinity to PBPs during transpeptidation. In conclusion, d-serine had synergistic activity in combination with -lactams against MRSA strains both and . Considering the relatively good safety of d-serine alone or in combination with -lactams, d-serine is worth following up as new anti-MRSA infection strategies.

Association between frailty and serum biomarkers in older adults / 中华老年医学杂志

Objective To investigate the relationship between frailty and serum biomarkers in the elderly. Methods A total of 371 elderly individuals aged 60 years and above with complete medical data were recruited during health examinations. Frailty phenotype assessment and comprehensive geriatric assessment were conducted.Serum levels of interleukin-6 (IL-6 ) ,high sensitivity C-reactive protein(hs-CRP) ,tumor necrosis factor-α(TNF-α) ,homocysteine(Hcy) ,insulin-like growth factor-1(IGF-1) ,25-hydroxyvitamin D[25(OH)D] ,folic acid and vitamin B12(VitB12) were detected by enzyme-linked immunosorbent assays ( ELISA ) and chemiluminescence immunoassays. Associations between frailty and the above factors were analyzed. Results Serum levels of IL-6 ,TNF-α ,Hcy and IGF-1 were significantly elevated along with progressive increase in frailty severity(all P＜0.05).There were a downward trend in serum 25(OH)D levels and an upward trend in serum hs-CRP ,folic acid and VitB12 levels as frailty severity increased ,but the changes did not amount to any statistical significance(all P＞0.05).Logistic regression analysis showed that ,after adjusting for age ,gender ,body mass index (BMI)and some clinical aspects (hearing loss ,urinary incontinence ,pain ,malnutrition ,cognitive dysfunction ,decreased activities of daily living ,depression , insomnia ,and anemia) ,serum levels of IL-6(OR=1.012 ,95% CI=1.005-2.041 ,P=0.033) ,IGF-1 (OR= 1.017 ,95% CI = 1.011-1.118 ,P= 0.021)and Hcy (OR= 1.007 ,95% CI :1.002-1.073 ,P=0.047)were significantly associated with frailty status. Conclusions Serum levels of IL-6 ,Hcy and IGF-1 are related to frailty status and may be used as potential biomarkers for the assessment of frailty in older adults.

Non-targeted metabolomics in septic mice infected with Klebsiella pneumoniae / 药学学报

UHPLC-QTOF-MS was applied to non-targeted metabolomics study of mice infected with K. pneumoniae ATCC® BAA 2146 to discover potential biomarkers and metabolic pathways that are associated with sepsis. Fifty-eight metabolites were identified by principal components analysis (PCA) and partial least-squares discriminant analysis (OPLS-DA), which was combined with variable projection importance (VIP) and nonparametric test. Eighteen of the 58 metabolites were further found to be involved in 8 metabolic pathways, including nicotinate and nicotinamide metabolism, pyrimidine metabolism, vitamin B6 metabolism, taurine and hypotaurine metabolism, arginine and proline metabolism, alanine, aspartate and glutamate metabolism, D-glutamine and D-glutamate metabolism and glycerophospholipid metabolism.

Risk of Active Pulmonary Tuberculosis among Patients with Coal Workers'Pneumoconiosis: A Case-control Study in China / 生物医学与环境科学（英文）

The aim of this study was to estimate the association between occupational, environmental, behavioral risk factors, and active pulmonary tuberculosis (PTB) among coal workers' pneumoconiosis (CWP) patients. A matched case-control study was conducted in 86 CWP patients with active PTB and 86 CWP controls without TB. A standardized questionnaire was used for risk factors assessment. Conditioned logistic regression analysis was used to identify associations between the risk factors and active PTB among CWP patients. The results showed that the stage of CWP, poor workplace ventilation, family history of TB, and exposure to TB were independent risk factors for active PTB in patients with CWP with which recommendations for improving work environments, and for case finding activities in patients with CWP could be made.

The application value of MDCE-MRI in the early chemoradiation efficacy in cervical cancer / 实用放射学杂志

Objective To explore the application value of multiphase dynamic contrast-enhanced MRI (MDCE-MRI) in the early chemoradiation efficacy in cervical cancer.Methods Twenty-one patients with cervical cancer received chemoradiation.Routine MR scan and MDCE-MRI scan were examined at different stages, including pre-treatment, 15 days,1 month and 2 months after treatment.According to volume change of the tumor after treatment, the cases were mainly divided into complete remission(CR), partial remission (PR).The correlation between volume change of cancer after treatment and the time-signal intensity curve (TIC),mean time to enhancement(MTE),time to peak(TTP),maximum slope of increase(MSI),maximum slope of decrease(MSD), negative enhancement integral (NEI) were analysed.Results After system chemoradiation for 2 months,4 of enrolled patients got complete remission and 17 partial remission.The reduction rate of the tumor diameter was negatively correlated to MSI values(r=-0.877,P<0.05) and positively related with NEI (r=0.819,P<0.05) before the treatment.15 days after the treatment, the diminished rate of the tumor diameter had active relation with change of TTP(r=0.765,P<0.05) and NEI(r=0.775,P<0.05).It indicated that MSI and NEI values before cervical cancer chemoradiation and the variation of TTP and NEI after treatment for 15 days could help predict the variation tendency of cancer diameter.Chemoradiation and therapy 1 month later,for all the parameters'' AUC values,the change rate of NEI were the largest one.When the critical value was not less than 96.46%, the sensitivity was 82.4%, specificity was 75%.TIC curve of type Ⅰ increased, type Ⅲ decreased to disappear gradually.Conclusion MDCE-MRI can be used to predict the efficacy of chemoradiation in cervical cancer patients, of which the MSI, TTP and NEI are more sensitive.

Analysis of the Processing Situation of Marketed Scorpions and Optimization of Salt-water Processing / 中国药师

Objective: To analyze the processing situation of 124 batches of scorpions and optimize the salt-water processing.Methods: The character, extract, moisture, total ash, acid insoluble ash, amino acids content and heavy metals in the marketed scorpions were determined.Using extract and amino acids contents as the indices, the salt-water processing was optimized.Results: The results of the above determinations showed that the quality difference was very obvious for the marketed scorpion, and the extract and amino acids contents were the highest in scorpion boiled in 2% sodium chloride aqueous solution for 5 min.Conclusion: The item of extract test should be revised in the current standard, and the items such as moisture, total ash, acid insoluble ash, amino acids content and heavy metals should be taken into consideration in order to control the quality of scorpion more comprehensively.The best salt-water processing of scorpion is boiled in 2% sodium chloride aqueous solution for 5 min.

In vitro metabolism and drug-drug interaction potential of IG-105, a novel antimicrotubule agent / 药学学报

IG-105, N-(2,6-dimethoxypyridine-3-yl)-9-methylcarbazole-3-sulfonamide, a novel antimicrotubule agent, showed potent anticancer activity in a variety of human tumor cells in vitro and in vivo. In order to characterize the metabolism and the possible drug-drug interaction of IG-105, we carried out a series of experiments. Drug metabolizing enzymes involved in IG-105 metabolism were investigated by using pooled human liver microsomes (HLMs) and recombinant cytochrome P450 isoforms (rP450s) respectively. The possible metabolites were analyzed by liquid chromatography-orbitrap-mass spectrometry (LC-Orbitrap-MS). The inhibitory effect of IG-105 on main P450 enzymes was also evaluated. The results showed that IG-105 can be metabolized by a series of rP450s, including CYP1A2, CYP2B6, CYP2C9, CYP2C19, CYP2D6, CYP3A4 and CYP3A5, with the major contribution enzymes being CYP1A2, CYP2B6, CYP2C19, and CYP3A. Three metabolites (M1-M3) were identified and demethylation was the major phase I metabolic reaction for IG-105. IG-105 moderately inhibited CYP1A2, CYP2C9, CYP2C19, CYP2D6, and CYP3A enzyme activities with IC50 values of 6.42, 23.64, 0.39, 1.4, and 3.14 μmol·L-1, respectively. Since the biotransformation of IG-105 involves multiple enzymatic pathways, the compound is less likely to be a victim of a concomitantly used medicine which inhibits activity of one of the CYPs. However, as IG-105 showed medium to strong inhibition on CYP1A2, CYP2D6, CYP3A, and CYP2C19, caution is particularly needed when IG-105 is co-administrated with other anticancer drugs which are mainly metabolized by the above enzymes.

Effects of Alprostadil and Yishen Huashi on Blood Glucose, Blood Lipid,Renal Function and Urinary Podocyte Proteins of Patients with Diabetic Nephropathy / 现代生物医学进展

Objective:To analysis the effects of alprostadil and yishen huashi particles on blood glucose,blood lipid,renal function and urinary podocyte proteins of patients with diabetic nephropathy.Methods:98 patients with diabetic nephropathy were selected and randomly divided into the control group and the experimental group with 49 cases in each group.The patients in the control group were treated with alprostadil,while the patients in the experimental group were treated with yishenhuashui particles on the basis of the control group.Then the curative effect,the levels of glycosylated hemoglobin (HbAlc),blood glucose (FPG),2 h postprandial blood glucose (2h PG),triglycerides (TG),total cholesterol (TC),high-density lipoprotein (HDL-C),low density lipoprotein (LDL-C),blood urea nitrogen (BUN),creatinine (Cr),β2 microglobulin (β2-MG),bladder inhibition (Cys-C) and urinary podocyte proteins (PCX) in the two groups were observed and compared between the two groups before and after the treatment.Results:The total effective rate of experimental group was higher than control group (P＜0.05).After treatment,there was no statistically significant difference about the HbA1c,FPG and 2 HPG between the two groups (P＞0.05).After the treatment,the levels ofTG,TC,LDL-C,BUN,Cr,β2 MG,Cys C,PCX and urinary nephrin/urine Cr of the experimental group were lower than those of the control group (P＜0.05).The HDL-C of experimental group was higher than that of the control group (P＜0.05).Conclusion:The curative is effect of alprostadil and yishen huashi particles in treatment diabetic nephropathy patients,can conducive to the improvement of blood glucose,blood lipid,renal function,reduce the concentration of urinary podocyte related proteins.

The progress in the treatment of fibrodysplasia ossificans progressiva / 中华内分泌代谢杂志

Fibrodysplasia ossificans progressiva(FOP) is a rare debilitating genetic disorder characterized by malformation of the great toes during embryonic skeletal development as well as progressive heterotopic ossification postnatally. There is no known definite treatment. Here presented a review of the recent advances in the potential FOP therapies.

Effect of IL-2 on primary hepatic carcinoma and its potential clinical value / 国际肿瘤学杂志

As a T cell subsets growth factor,interleukin-2 (IL-2) can inhibit cancer cell proliferation,reduce tumorigenicity and transitivity,improve local tumor microenvironment,enhance the immunogenicity of hepatocellular carcinoma cells,reduce the load of hepatitis virus and greatly enhance the ability of anti-tumor.On the other hand,it can enhance tumor hypercoagulable state.The effect of IL-2 on primary hepatic carcinoma and its potential clinical value have a bright future.

Validated LC--MS/MS method for determination of YH-8, a novel PKnB inhibitor, in rat plasma and its application to pharmacokinetic study

(E)-Methyl-4-aryl-4-oxabut-2-enoate (YH-8) is a novel PKnB protein kinase inhibitor with good anti-tuberculosis activity. To evaluate its pharmacokinetics in rats, a sensitive and selective high performance liquid chromatography-tandem mass spectrometric (LC--MS/MS) method has been developed and validated for the quantification of YH-8 in rat plasma for the first time. Samples were pre-treated using a liquid--liquid extraction with ethyl acetate and the chromatographic separation was performed on a C18 column by gradient elution with methanol--water as the mobile phase. YH-8 was detected using a tandem mass spectrometer in positive selected reaction monitoring (SRM) mode. Method validation revealed good linearity over the range of 1-500 ng/mL for YH-8 with a lower limit of quantification (LLOQ) of 1 ng/mL. Intra- and inter-day precision of YH-8 assay in rat plasma samples were 2.0%-6.8%, with accuracy of the method being 100.69%-106.18%. Stability test showed that when spiked into rat plasma, YH-8 was stable for 12 h at room temperature, for up to 15 days at -70 °C, and after three freeze-thaw cycles. Extracted samples were found to be stable over 12 h in an auto-sampler. The method was successfully applied to the pharmacokinetic study of YH-8 in rats after oral administration at 100 mg/kg and 200 mg/kg.

Mechanism of T helper cell for demyelinated optic neuritis / 中华实验眼科杂志

Background Optic neuritis is one of the common clinical neuro-ophthalmic diseases.Recurrent optic neuritis can cause irreversible axonal damage and visual impairment,and no prevention method is effective up to now.Its pathogenesis is considered to be associated with the body's immune imbalance.Objective This proposal tried to establish experimental autoimmune encephalomyelitis (EAE) and explore the immunomodulatory effects of T helper (Treg),Th17 cell in the model of optic neuritis.Make the immune mechanism of demyelinating optic neuritis clear.Methods EAE model was established to observe the clinical symptoms and visual electrophysiological changes of optical neuritis in mice;hematoxylin-eosin staining to observe the histological changes in the mouse models of EAE; immunohistochemical staining to detect optic nerve axon damage markers β-amyloid peptide precurso (β-APP) protein expression quantity changes;reverse transcription PCR (RT-PCR) to detect the dynamic expression of forkhead/winged helix transcription factor p3 (Foxp3),transforming growth factor-β (TGF-β),interleukin-1β (IL-1β),IL-17,IL-10 in optic nerve tissue.Results After immunization 11 days,neurological symptoms of EAE mice started to appear,hematoxylin-eosin staining showed optic nerve tissue infiltration of inflammatory cytokines; immunohistochemical staining showed enhancement of axonal injury markers β-APP positive staining; flash visual evoked potential(F-VEP) test showed 7,11,14,19,23,28 days,compared with the normal mice,P1 prepatents of model group were prolonged after modeling,the differences were statistically significant (t =4.487,15.203,16.364,11.540,11.959,16.163,all at P＜0.05).When 7 days after modeling,the difference of N1-P1 amplitude between normal group and model group was no statistical significance (t =-0.992,P =0.378); while 11,14,19,23,28 days after modeling,compared with normal mice,N1-P1 amplitudes of model mice were significantly lower,the differences were statistically significant(all at P＜ 0.05).The expressions of TGF-β,IL-1β,IL-17,Foxp3,IL-10 mRNA among different time points had stastistie significant differences (F =12.721,15.015,14.343,69.374,68.290,all at P =0.000),compared with the normal group,when 11 days,14 days after modeling,TGF-β mRNA levels were significantly increased ; 19 days,23 days after modeling,IL-1β mRNA levels were significantly increased;7 days,11 days after modeling,IL-17 mRNA levels were significantly increased ;7,11,14,19,23,28 days after modeling,Foxp3 mRNA levels were significantly lower; 19,23,28 days after modeling,IL-10 mRNA levels were significantly lower,the difference were statistically significant (all at P＜ 0.05).Conclusions Th17 cells launch demyelinating optic neuritis immune injury,and Th1 subgroup performs the function of maintaining inflammatory lesions,Treg cell subgroup is abnormal much earlier than that of the Th2 subgroup.The imbalance of Th17/Treg cell may be involved in the pathogenesis of demyelinating optic neuritis.