OBJECTIVE: To establish the method to analyze γ-hydroxybutyric acid (GHB) and its precursors 1,4-butanediol (1,4-BD) and gamma-butyrolactone (GBL) in urine through LC-MS/MS and provide evidence for related cases. METHODS: GHB-d6 and MOR-d3 were used as the internal standard. The urine sample was separated by LC after protein precipitation with methanol. The electrospray ion source was for ionization. Each compound was detected through multiple-reaction monitoring (MRM) mode. RESULTS: The limits of detection of GHB and its precursors 1,4-BD and GBL were 0.1, 0.1 and 2 μg/mL. The accuracy was 87.6%-98.1%. The intra-day and inter-day precisions were less than 15% and matrix effects were higher than 80%. CONCLUSION The method is high sensitive, simple, rapid, specific and with high reliability. This study has provided technical support and basic data for forensic cases involving GHB.
4-Butyrolactone/urine* ; Butylene Glycols/urine* ; Chromatography, Liquid ; Forensic Sciences ; Humans ; Hydroxybutyrates/urine* ; Mass Spectrometry ; Reproducibility of Results ; Tandem Mass Spectrometry

Aim To ascertain whether defibrase has the protective effect against reperfusion injury after cerebral ischemia.Methods 70 renovascular hypertensive rats(RHR) were randomly divided into defibrase group, control group and sham-operated group.Reversible middle cerebral artery occlusion(MCAO) models were produced by the modified. Longa's method,and reperfusion was begun 2 hours after occlusion.Rats in the defibrase group were given defibrase 10 U?kg-1 body weight via femonal intraveneous injection, and in the control group with the same amount of saline. The brain pieces were processed by TTC and HE staining and the infarct size,brain microvessels damage and secondary bleeding were compared between the two groups. Results The volume of infarction in the defibrase group was obviously smaller than in the control group, the damage of brain microvessels was less severe, and the bleeding lesions under optical microscope were less than in the control group. Conclusion Defibrase has protective effect against reperfusion injury post cerebral ischemia.

OBJECTIVETo observe the clinical efficacy of treating chronic persistent bronchial asthma (CPBA) children with abnormal myocardial enzyme spectrum (AMES) by Yupingfeng Powder (YP) combined routine therapy.

METHODSFrom January 2010 to December 2012, 156 CPBA children patients with AMES were randomly assigned to the treatment group (80 cases) and the control group (76 cases). All patients received routine treatment (inhaled corticosteroids and/or leukotriene regulator). Besides, those in the treatment group took YP. The treatment duration was 3 months. The scores of children asthma control test (C-ACT), pulmonary function (FEV,% and PEF%), myocardial enzyme spectrum were observed before and after treatment, and 3 months before and after treatment. The myocardial enzyme spectrum of 40 healthy children at the baby clinics during the same period were recruited as the control.

RESULTSCompared with the control group, creatine kinase isoenzyme (CK-MB), creatine kinase(CK), and lactate dehydrogenase (LDH) increased in the two treatment groups (P <0.01), but there was no statistical difference in AST (P >0.05). Compared with before treatment in the same group, CK-MB, CK, LDH, and AST decreased in the treatment group after treatment and 3 months after treatment (P <0.01). CK-MB, CK, LDH, and AST decreased in the control group 3 months after treatment (P <0.01, P <0.05).Compared with after treatment, CK decreased in the control group 3 months after treatment (P <0.01). C-ACT score, FEV(1),%, and PEF% all increased in the two groups after treatment and 3 months after treatment (P <0.01, P <0.05). Compared with after treatment in the same group, CK decreased in the control group 3 months after treatment (P <0. 01). Compared with the control group in the same period, post-treatment CK-MB and CK decreased (P <0. 01, P <0. 05), while post-treatment C-ACT score, FEV, %, and PEF% increased (P <0.05) in the treatment group (P <0.05).

CONCLUSIONYP could strengthen specific and non-specific immunity of the organism, and improve clinical symptoms and the level of myocardial enzyme spectrum.

Asthma ; therapy ; Child ; Chronic Disease ; therapy ; Creatine Kinase, MB Form ; metabolism ; Drugs, Chinese Herbal ; therapeutic use ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Myocardium ; enzymology

The genes of maltooligosyl trehalose synthase (MTSase) and maltooligosyl trehalose tetrahydrolase(MTHase) from Sulfolobus solfataricus ATCC 35092 were amplified using PCR. The expression plasmids, pTrc99a-MTSase and pTrc99a-MTHase, were constructed by inserting these two DNA fragments into E. coli expression vector pTrc99a. The specific activity of MTSase and MTHase in E. coli BL21(DE3) at optimal fermentation conditions reached 31.3U/g (wet cell) and 403U/g (wet cell), respectively. The biotransformation of partially hydrolyzed starch to trehalose catalyzed by MTSase and MTHase was carried out at 75℃ and pH 5.0. The highest yield of trehalose (ca. 53.6%) was gained when the original starch concentration was 15%(w/v) and the DE value was 10.

Objective:To investigate the influencing factors of the therapeutic effect of epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) in patients with epidermal growth factor receptor (EGFR) mutant advanced non-small cell lung cancer (NSCLC).Methods:The clinical data of 104 EGFR mutant advanced NSCLC patients who received EGFR-TKI treatment in Wuxi No.2 Hospital Affiliated to Nanjing Medical University from January 2015 to October 2019 were collected. The correlation of different types of EGFR mutation with the clinicopathological characteristics, the hematological examination results and the treatment mode of patients was analyzed. Cox proportional hazard model was used to analyze the association of the progression-free survival (PFS) time of patients receiving EGFR-TKI treatment with the different types of EGFR mutation, the clinicopathological characteristics, hematological related indexes and treatment mode. Kaplan-Meier method was used to analyze the independent influencing factors for the PFS of the stratified patients.Results:The overall disease control rate (DCR) of patients receiving EGFR-TKI treatment was 92.3% (96/104). Cox univariate analysis showed that the levels of carcinoembryonic antigen (CEA), carbohydrate antigen 125 (CA125), D-dimer, and previous surgical treatment history of patients receiving EGFR-TKI treatment were associated with PFS of patients (all P < 0.05). Cox multi-factor analysis showed that EGFR mutation type ( HR =2.371, 95% CI 1.298-4.332, P = 0.005), combination therapy ( HR = 0.489, 95% CI 0.245-0.978, P = 0.043) and choice of therapeutic drugs ( HR = 0.261, 95% CI 0.113-0.606, P = 0.002) were independent influencing factors for the PFS of patients receiving EGFR-TKI treatment. The PFS of EGFR 19 exon-mutant patients with advanced NSCLC was longer than that of those with EGFR 21 exon-mutant (median PFS time: 14.0 months vs.9.5 months, P<0.05); the PFS of combination of radiotherapy or chemotherapy was longer than that of EGFR-TKI single therapy (median PFS time: 15.0 months vs. 9.0 months, P<0.05), the PFS of patients receiving erlotinib was better than that of those receiving gefitinib ( P<0.05). According to EGFR mutation types, it was found that EGFR 19 exon-mutant patients receiving EGFR-TKI in first-line treatment could obtain better PFS than those who receiving EGFR-TKI in second-line and above treatment (median PFS time: 14.0 months vs. 9.5 months, P<0.05). When receiving EGFR-TKI, EGFR 19 exon-mutant patients with CA125 <85 U/ml could obtain longer PFS time than those with CA125 ≥85 U/ml (median PFS time: 14.0 months vs. 6.5 months, P<0.05). Conclusions:The therapeutic effect of EGFR-TKI in EGFR-mutant patients with advanced NSCLC is positive. EGFR 19 exon-mutant NSCLC patients with low-level CA125 receiving EGFR-TKI in first-line treatment can obtain better PFS.

AIM: To test the ability of multifocal visual evoked potential (mfVEP) in the detecting of glaucoma by comparing the mfVEP recorded from normal subjects and glaucoma patients.METHODS: The mfVEP of 32 normal eyes (n =21) and of 58 eyes (n =37) with primary glaucoma were recorded with the Vision Monitor electrophysical apparatus by the second kernel analysis and to determine the correlation of the topographic location between them.RESULTS: There were significant variability (the coefficient of variation was 43.05%) in mfVEP RMS amplitude in the normal subjects; The RMS amplitude of eyes with glaucoma were smaller than that of the normal eyes and significantly statistical difference were found in the relatively center (namely the 0° -10° ring zone) and in superior nasal quadrant (P＜0.05) while there were no significantly statistical differences of the latency time between them.CONCLUSION: The normal subjects have large individual variability of mfVEP responses. The RMS amplitude of the mfVEP of glaucomatous eyes descends, especially in center zone and superior nasal quadrant.

NMDA-induced excitotoxicity cause severe neuronal damage including apoptosis and necrosis. The present study was aimed to evaluate the proportion of NMDA-induced apoptosis of rat cortical neurons and discover signal transduction mechanism. Caspase inhibitor and lactate dehydrogenase (LDH) assay were used to study the NMDA-induced apoptosis. To explore the involved signal pathways, the primary culture of rat cortical neurons were pretreated by the inhibitors of three MAPK pathways, extracellular signal-regulated kinase (ERK), c-Jun N-terminal kinase (JNK), and p38 MAPK. With 2 h of NMDA treatment, cellular apoptosis was measured by caspase-3 activity, terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick-end labeling (TUNEL) and Annexin V staining. The results showed that: (1) Caspase-dependent apoptosis accounted for 22.49% in NMDA-induced neuronal death; (2) Pretreatment with p38 MAPK inhibitor SB203580 (10 μmol/L) significantly decreased NMDA-mediated caspase-3 activity by 30.43% (P < 0.05). However, ERK inhibitor PD98059 (20 μmol/L) or JNK inhibitor SP600125 (20 μmol/L) did not influence caspase-3 activity; (3) Pretreatment with SB203580 significantly reduced the number of NMDA-induced TUNEL-positive cells by 33.10% (P < 0.05). PD98059 (20 μmol/L) or SP600125 (20 μmol/L) did not show obvious effect; (4) Pretreatment with SB203580 (10 μmol/L) significantly reduced the number of NMDA-induced early apoptotic neurons by 55.56% (P < 0.05). Also, SP600125 (20 μmol/L) significantly decreased the amount of late apoptotic/dead cells by 67.59% (P < 0.05). There was no effect of PD98059 (20 μmol/L). These results indicate that: (1) NMDA induces neuronal apoptosis besides necrosis; (2) p38 MAPK, but not JNK and ERK, is involved in NMDA-induced neuronal apoptosis, and inhibition of the apoptotic signaling pathway contributes to neuroprotection; (3) JNK activation might contribute to NMDA-induced neuronal necrosis rather than apoptosis.
Animals ; Anthracenes ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Cells, Cultured ; Extracellular Signal-Regulated MAP Kinases ; antagonists & inhibitors ; Imidazoles ; pharmacology ; JNK Mitogen-Activated Protein Kinases ; antagonists & inhibitors ; MAP Kinase Signaling System ; N-Methylaspartate ; pharmacology ; Neurons ; cytology ; Primary Cell Culture ; Pyridines ; pharmacology ; Rats ; p38 Mitogen-Activated Protein Kinases ; antagonists & inhibitors

Adult ; China ; epidemiology ; Female ; Herpesviridae Infections ; epidemiology ; Herpesvirus 8, Human ; Humans ; Male ; Middle Aged ; Seroepidemiologic Studies ; Substance-Related Disorders ; epidemiology ; virology ; Young Adult

Background Corneal epithelial abrasion results in corneal ulcer and stroma cloudy evenb irreversible visual impairment.Previous drugs for corneal epithelial injury can only alleviate the inflammatory irritation.So it is very important to seek a drug which regulate the growth of corneal epithelium.Objective This study was to investigate the effects of recombinant human BIGH3 protein eye drops on corneal epithelial abrasion.Methods Fifty right eyes of 50 clean adult New Zealand white rabbits were collected.Two rabbits were sacrificed right away following establishment of corneal epithelial abrasion models (0 hour group).The other 48 rabbits were randomly divided into recombinant human epidermal growth factor (EGF) derivative group (positive control group),normal saline solution group (negative control group),0.25％ or 0.5％ recombinant human BIGH3 protein eye drops group.Corneal abrasion models were created with alcohol corrosion method with a defect area of 7 mm2.The corresponding eye drops were used separately in 4 groups for four times per day after operation.Experimental eyes were examined by the slit lamp microscope,and fluorescein vital staining were performed 12,24,36,48,72 hours after operation.Planimetry was performed and the corneal photographs were analyzed with computer software.The rabbits were sacrificed 12,24,36,48 and 72 hours after operation,respectively,and the histopathological examination of corneal tissue was carried out.Results No obvious irritation response was seen after administered of eye drops in the recombinant human EGF derivative group,normal saline solution group,0.25％ and 0.5％ recombinant human BIGH3 protein eye drops groups.Histopathological examination revealed a full-thickness defect of corneal epithelium after modeling.The defect area was gradually smaller with time lapse,and corneal epithelium migrated from periphery toward the center zone.Corneal epithelial cells increased with time lapse.Compared with normal saline solution group,the defect area of corneal epithelium lessened 12,24,36,48 hours after operation in the 0.25％,0.5％ recombinant human BIGH3 protein eye drops groups and recombinant human EGF derivative group (all at P =0.000),but at 12and 24,36 hours after operation,no significant differences were found between the recombinant human EGF derivative group and normal saline solution group (P =0.321,0.057,0.126).The defect area was smaller in the 0.5％recombinant human BIGH3 protein eye drops group than that of the recombinant human EGF derivative group at various time points (P=0.042,0.039,0.025,0.008).However,significant smaller defect area was exhibited only at 12 hours and 24 hours after operation in the 0.25％ recombinant human BIGH3 protein eye drops group (P=0.047,0.042).No significant differences were seen in corneal defect area at various time points between 0.25％ and 0.5％recombinant human BIGH3 protein eye drops groups (P =0.358,0.259,0.108,0.062).In addition,the corneal defect area was (0.51 ±0.42)mm2 72 hours after operation in the normal saline group;while that in the recombinant human EGF derivative group and recombinant human BIGH3 protein eye drops groups was disappeared.The repairing curves in the recombinant human BIGH3 protein eye drops groups were superior to those of the recombinant human EGF derivative group and normal saline solution group.Conclusions 0.25％ and 0.5％ recombinant human BIGH3 protein eye drops have facilitation effect on the growth of corneal epithelial cells and the healing of corneal injury.

OBJECTIVETo research the co-infections of HIV and human herpesvirus 8 (HHV8) in Uygur high-risk groups of HIV infection in a city Xinjiang.

METHODSAll 468 Uygurs at high HIV risk registered in the sentinel monitoring system in 2006 were enrolled in this study. The antibodies to HHV8 latency-associated nuclear antigens 1 (LANA1), lytic antigens open reading frame 65 (ORF65) and K8.1 were measured by enzyme linked immunosorbent assay (ELISA). Chi-square test and non-condition Logistic regression model were used for data analysis.

RESULTSOf 468 sera samples, 67 (14.3%) were HIV and HHV8 co-infection positive.Male's HIV and HHV8 co-infection rate (22.6%, 54/239) was higher than the female's (5.7%, 13/229) (chi(2) = 27.285, P < 0.001). For those above 24 year old, HIV and HHV8 co-infection rate (15.8%, 65/412) was higher than the < 24 year old group's (3.6%, 2/56) (chi(2) = 5.987, P = 0.014). The group of Elementary school and illiterate people's HIV and HHV8 co-infection rate (20.7%, 40/193) was higher than the junior middle school and the above culture (9.8%, 27/275) (chi(2) = 10.999, P = 0.001). For the unmarried people, the co-infecting rate of HIV and HHV8 for the married, the cohabitants, the divorced or the widowers were 16.9% (14/83), 12.2% (42/345), 27.5% (11/40) respectively. There was significantly statistical difference among three marital status (chi(2) = 7.399, P = 0.025). Injecting drug users' HIV and HHV8 co-infection rate (26.5%, 50/189) was higher than non-injecting drug users' (6.1%, 17/279) (chi(2) = 38.083, P < 0.001), and stratified by gender, OR(M-H) was 4.207 (95%CI: 1.529 - 11.578). Via non-condition logistic stepwise regression analysis, only injecting drug use entered model, compared with non-injecting drug users, injecting drug users were more dangerous for HIV and HHV8 co-infecting (OR = 5.544; 95%CI: 3.081 - 9.975).

CONCLUSIONThe HIV and HHV8 co-infection rate was higher in the Uygurs at high HIV risk in Xinjiang. Injecting drug use is a risk factor of the HIV and HHV8 co-infection, which might be one of routes of HIV and HHV8 co-infection among this group.

Adolescent ; Adult ; Aged ; Child ; China ; epidemiology ; Female ; HIV Infections ; epidemiology ; ethnology ; virology ; Herpesviridae Infections ; epidemiology ; Herpesvirus 8, Human ; isolation & purification ; Humans ; Male ; Middle Aged ; Population Surveillance ; Young Adult