Streptococcus pneumoniae(S.pn) is an opportunity pathogenic bacteria,environmental factors play a key role in the pathogenicity of S.pn. It is important to study virulent gene in vivo. The S.pn suicide plasmid containing gfp reporter was constructed by fusing the genes pneumolysin and gfp,in which gfp is an excellent molecule probe in vivo. The plasmid was integrated to No.22 S.pn by homologous recombination. The recombinant S.pn was gained and evaluated in aspects of fluorescence excitation, biological character and physio-activity. The results showed it is efficient and available to report the expression of virulent genes in vivo and in vitro, which will provide a new easy method for evaluating and screening the virulent genes of S.pn in vivo.

Objective To observe the effect of power-assisted functional electrical stimulation (PAFES)therapy on ankle joint function recovery in stroke patients.Methods Ninety hemiplegic stroke patients were randomly and evenly divided into a control group,a PAFES group,and a neuromuscular electrical stimulation (NMES)group.All groups received conventional rehabilitation training.PAFES group adopted PAFES treatment on affected lower extremities and NMES group was given the NMES therapy on the tibialis anterior of the affected lower limbs,in addition to conventional rehabilitation training.The active range of motion (AROM) of ankle dorsiflexion,FuglMeyer motor assessment (FMA),Barthel index (BI) and Ankle flexion and extension movement (AFEM) in 10 seconds were evaluated before the trial and after 4 weeks treatment.Results After treatment,there were significant differences in the AROM of ankle dorsiflexion,FMA,BI and AFEM (P ＜ 0.05) compared with before treatment within each group.The improvement of AROM of ankle dorsiflexion in PAFES group (8.19 ± 3.39) ° and the values in NMES group (8.96 ± 3.68) ° were to a significantly greater extend than control group (3.88 ± 4.10) ° (P ＜0.05) ; the improvement of FMA and BI in PAFES group was also superior to those in NMES group and control group (P ＜ 0.05).Conclusion The intelligent PAFES therapy could help improve AROM of ankle dorsiflexion,the motor function of the affected lower extremity and the ability of the activities of daily living in stroke patients.

Reactive oxygen species(ROS) in the cardiac muscle are increased in cardiovascular diseases such as hypertension,coronary heart disease and heart failure.Oxidative stress mediates myocyte hypertrophy,myocyte apoptosis and interstitial fibrosis.And these changes are particularly pronounced in elderly patients or senescent animal disease models.The major sources of ROS in myocardium are mitochondria and the reduced nicotinamide adenine dinucleotide phosphate(NADPH) oxidase.Specific ROS such as hydrogen peroxide play an important role in myocardial remodeling in heart failure.Oxidative modifications of calcium handling proteins result in reduction of their activities,leading to decreased myocardial contractility.Reduction of oxidant levels and inhibition of oxidative modifications of specific proteins through inhibiting ROS sources may provide new strategies for the treatment of myocardial remodeling.

Objective To investigate the levels of serum vascular endothelial growth factor(VEGF) and basic fibroblast growth factor(bFGF) in patients with non-small cell lung cancer(NSCLC) and relationships with c1inicopatho1ogica1 characteristics and their clinical significance. Methods The concentrations of serum VEGF and bFGF were detected by enzyme-linked immunosorbent assay(ELISA) in 40 patients with NSCLC before and after chemotherapy. Results The level of serum VEGF in patients with Ⅳ stage NSCLC was significantly higher than that of Ⅲ stage(P

The coronavirus disease 2019 (COVID-19) is an acute infectious disease caused by the new severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, which has led to serious worldwide economic burden. Due to the continuous emergence of variants, vaccines and monoclonal antibodies are only partial effective against infections caused by distinct strains of SARS-CoV-2. Therefore, it is still of great importance to call for the development of broad-spectrum and effective small molecule drugs to combat both current and future outbreaks triggered by SARS-CoV-2. Cathepsin L (CatL) cleaves the spike glycoprotein (S) of SARS-CoV-2, playing an indispensable role in enhancing virus entry into host cells. Therefore CatL is one of the ideal targets for the development of pan-coronavirus inhibitor-based drugs. In this study, a CatL enzyme inhibitor screening model was established based on fluorescein labeled substrate. Two CatL inhibitors IMB 6290 and IMB 8014 with low cytotoxicity were obtained through high-throughput screening, the half inhibition concentrations (IC₅₀) of which were 11.53 ± 0.68 and 1.56 ± 1.10 μmol·L^-1, respectively. SDS-PAGE and cell-cell fusion experiments confirmed that the compounds inhibited the hydrolysis of S protein by CatL in a concentration-dependent manner. Surface plasmon resonance (SPR) detection showed that both compounds exhibited moderate binding affinity with CatL. Molecular docking revealed the binding mode between the compound and the CatL active pocket. The pseudovirus experiment further confirmed the inhibitory effects of IMB 8014 on the S protein mediated entry process. In vitro pharmacokinetic evaluation indicated that the compounds had relatively good drug-likeness properties. Our research suggested that these two compounds have the potential to be further developed as antiviral drugs for COVID-19 treatment.

Objective To evaluate the effects of OPRM1All8G and CYP3A4*18B genetic polymorphism and the interaction on postoperative analgesia with fentanyl in the patients undergoing radical resection of lung cancer.Methods One hundred and thirty-nine patients (native of Henan province),aged 40-64 yr,weighing 40-70 kg,of American Society of Anesthesiologists physical status Ⅰ or Ⅱ,scheduled for elective radical resection of lung cancer under general anesthesia,were enrolled in this study.The polymorphic sites of the OPRM1All8G and CYP3A4*18B allele were analyzed by using polymerase chain reaction technique and ABI 3130 Genetic Analyzer.The patients were divided into wild homozygote group (group AA,group *1/*1),heterozygote group (group AG,group * 1/*18B) and mutation homozygote group (group GG,group *18B/*1SB) according to their genotypes.The patients were divided into 7 groups according to the interaction between the two genes:AA plus *1/*1 group (group Ⅰ),AA plus *1/*18B group (group Ⅱ),AG plus *1/*1 group (group Ⅲ),AG plus *1/*18B group (group Ⅳ),GG plus * 1/*1 group (group Ⅴ),GG plus *1/*18B group (group Ⅵ) and *18B/*18B group (group Ⅶ).Patientcontrolled intravenous analgesia with fentanyl was started at the end of surgery to maintain the visual analogue scale ≤ 3 points.The amount of fentanyl used within 24 and 48 h after surgery was recorded,and the occurrence of adverse reactions within 48 h after surgery was observed.Results The amount of fentanyl used within 24 and 48 h after surgery was significantly higher in group GG than in group AA (P＜0.05).The amount of fentanyl used within 48 h after surgery was significantly lower in group *18B/*18B than in group *1/*1 (P＜0.05).The amount of fentanyl used within 48 h after surgery was significantly higher in Ⅱ and Ⅳ groups than in group Ⅰ,in group Ⅲ than in group Ⅱ,in group Ⅴ than in Ⅰ-Ⅳ groups,and in group Ⅵ than in Ⅱ and Ⅳ groups,and was significantly lower in group Ⅶ than in Ⅰ-Ⅵ groups (P＜ 0.05).There was no significant difference in the incidence of adverse reactions within 48 h after surgery between groups (P＞0.05).Conclusion OPRM1A1l8G and CYP3A4*18B genetic polymorphism and the interaction are the genetic factors contributing to individual variation in fentanyl pharmacodynamics in the patients undergoing radical resection of lung cancer.

Objective:To explore the mechanism,clinical features,and prognosis of trigemino-cardiac reflex (TCR) during skull base operations.Methods:A retrospective analysis was performed on 291 patients with skull base disease from Jan.2009 to Oct.2015 in Peking University First Hospital.By reviewing the patients' operative data and anaesthetic electrical record,and we picked out all the patients who suffered from TCR during the operation and analyzed their surgical procedures,clinical features,influence factors,and prognosis.TCR was defined as a drop in mean arterial blood pressure (MABP) and the heart rate (HR) of more than 20％ to the baseline values before the stimulus and coinciding with the manipulation of the trigeminal nerve.Results:In all the 291 patients receiving skull base surgery,9 patients suffering TCR for 19 times during the operation were found.These 9 cases included three acoustice schwannomas,one trigeminal schwannoma,one petroclival meningioma,one epidermoid cyst in cerebellopontine angle,one cavernous sinus cavernous hemangioma,one pituitary adenoma,and one trigeminal neuralgia.The trigger of TCR was related to manipulation,retraction,and stimulation of the trunk or branches of trigeminal nerve.The baseline heart rate was 62-119/min [mean (79.4 ± 14.6) /min] and dropped about 29.0％-66.4％ (mean 44.3％) to 22-60/min [mean (44.2 ±9.6) /min] after TCR.The baseline mean arterial blood pressure was 75-103 mmHg [mean (87.5 ±7.8) mmHg] and dropped about 23.4％-47.2％ (mean 37.3 ％) to 45-67 mmHg [mean (54.9 ± 6.3) mmHg] after TCR.During the 19 times of TCR,heart rate and blood pressure could return to baseline in a short time while stopping manipulation (8 times),using atropine (8 times,dose 0.5-1.0 mg,mean 0.69 mg),using ephedrine hydrochloride (one time,15 mg),using epinephrine (one time,1 mg),and using dopamine (one time,2 mg).TCR also could be triggered again by a second stimulation.There was no relative cardiologic complication or neurological deficit in the postoperative 24 hours.Conclusion:TCR is a short neural reflex with a drop in blood pressure and heart rate coinciding with the manipulation of the trigeminal nerve in skull base surgery.Correct recognition,intensive observation,and essential management of TCR will lead to a good prognosis.

Eleven C21 steroids were isolated from chloroform extract of roots of Cynanchum otophyllumby silica gel, MCI, ODS columns, and semi-preparative HPLC. Their structures were determined by spectroscopic data analysis as otophylloside B(1), caudatin-3-O-beta-D-cymaropyranosyl-(1-->4)-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside (2), caudatin-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside (3), caudatin-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-digitoxopyranosyl-(1-->4)-beta-D-cymaropyranoside (4), otophylloside O (5), gagamine-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-cymaropyranoside (6), sinomarinoside B (7), mucronatosides C (8), wallicoside J (9), stephanoside H (10), and qinyangshengenin-3-O-beta-D-oleandropyranosyl-(1-->4)-beta-D-cymaropyranosyl-(1-->4)-beta-D-digitoxopyranoside (11). Among them, compounds 2-3, and 6-11 were separated from the roots of this plant for the first time.
Cynanchum ; chemistry ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Plant Roots ; chemistry ; Steroids ; chemistry ; isolation & purification

OBJECTIVETo analyze the relation of plasma D-dimer levels and incidence of deep venous thrombosis after spinal surgery.

METHODSThe clinical data of 63 patients underwent spinal surgery from October 2009 to October 2010 were retrospective analyzed. There were 40 males and 23 females with an average age of 48 years old(21 to 76) in operation. Operation levels of 15 cases were in cervical vertebrae, 4 cases were in thoracic vertebrae,and 44 cases were in lumbar vertebrae. Thirty patients with spinal fracture were caused by trauma and 33 patients without trauma, 11 patients combined with nerve injury. The patients were divided into two groups according to plasma D-dimer levels, more than or equal to 500 microg/L was D-dimer positive group and less than 500 microg/L was D-dimer negative group. Venous blood of all patients early morning with empty stomach were testd on admission, and at 2 h, 1 d, 2 d, 3 d, 4 d, 6 d, 8 d, 10 d, 15 d after operation,respectively.

RESULTSThere was no statistically significant differences in sex, operative segments, implants, operative posture, age, bleed volume, body weight, peroperative D-dimer levels between two groups. After operation, plasma D-dimer of 19 patients were more than or equal to 500 microg/L, with persistent or progressive increasing. Two cases occurred deep venous thrombosis in D-dimer positive group, they respectively were found at 3 days and 8 days after operation. Both of them underwent posterior decompression and internal fixation. However,no deep venous thrombosis was found in D-dimer negative group.

CONCLUSIONPostoperative D-dimer assay can effective predict deep venous thrombosis occurrence. D-dimer level more than or equal to 500 microg/L will be considered as a risk factor for deep venous thrombosis after spinal surgery.

Adult ; Aged ; Female ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Spine ; surgery ; Ultrasonography ; Venous Thrombosis ; blood ; diagnostic imaging ; surgery ; Young Adult