OBJECTIVETo establish a method for determination of 2,5-hexanedione in urine by headspace solid-phase microextraction-gas chromatography.

METHODSAfter extraction by solid-phase microextraction head, 2,5-hexanedione in urine was determined by gas chromatography and was quantified by external standard method.

RESULTSThe concentration of 2,5-hexanedione in urine showed a linear relationship within the range of 0.1-20.0 µg/ml. The regression equation was y=261.36x-1.903 3, r=0.999 2. The minimum detectable concentration was 0.01 µg/ml. The recovery rate was 92.6%-97.1%, with a relative standard deviation (RSD) of 3.3%-5.8%. The intra-day and inter-day RSDs were 3.8%-6.2% and 4.7%-6.3% respectively.

CONCLUSIONThis determination method has no requirement for organic solvents, features simple and rapid operation, possesses higher detection sensitivity, and applies well to the determination of 2,5-hexanedione in urine.

Objective: To establish a method for the determination of diethyl phthalate by gas chromatography in the air of workplaces. Methods: Diethyl phthalate in the air of workplace was collected throμgh glass fiber filter, eluted with methylbenzene, and detected by gas chromatography coupled with FID detectors. Results: The linear range of diethyl phthalate determined by this method was 14.0~1 400 μg/ml, y=2.09801x-3.66229, and the coefficient correlation was 0.999 99. The detection limit was 1.10 μg/ml, and the minimum detection concentration was 0.18mg/m³ (collected sample volume was 30 L) . The within-run precisions were 1.04%~2.75%, and the between-run precisions were 0.34%~1.30%. The recovery rates were 98.72%~103.21%, and sampling efficiency was 97.2%~100.0%. The elution efficiencies were 97.25%~98.68%. The samples could be stored at room temperature for 15 days. Conclusion The indicators established in this study were conformed with the requirements of GBZ/T210.4-2008, "The Guidelines for the Development of Occupational Hygiene Standards Methods Part 4: Determination of Chemical Substances in the Air of Workplaces" . Diethyl phthalate in the workplace air could be rapidly collected, accurate separated and determinated. This method is applicable to the determination of diethyl phthalate in the workplace air.

Objective: To establish a practical method forsampling and detectingtributyl phosphate intheworkplace. Methods: The samples were extracted by glass fiber membrane, eluted with ether, separated by gas chromatography, and detected by flame photometric detector. Results: There were good linear relationship in the minimum detection concentration was 7.2-720.0 μg/ml, and the correlation coefficient was 0.999 92. The detection limit was 0.86 μg/ml, and the minimum detection concentration was 0.14 mg/m³ (sample volume was 30 L) . Recovery rates were 99.8%-100.2%. The with-in relative standard deviations were 4.0%-5.4% and the between relative standard deviations were 2.0%-5.5%, and average samplingefficiency was about 99.1%-100.0%. Conclusion This method conforms with the requirements of "Standardization of Methods for Determination of Toxic Substance in Workplace" . Tributyl phosphate in air could be determined accurately using this method.

Objective To establish a method for determination of 2,5-hexanedione in urine by headspace solid-phase microextraction-gas chromatography.Methods After extraction by solid-phase microextraction head,2,5-hexanedione in urine was determined by gas chromatography and was quantified by external standard method.Results The concentration of 2,5-hexanedione in urine showed a linear relationship within the range of 0.1-20.0 μg/ml.The regression equation was y=261.36x-1.903 3,r=0.999 2.The minimum detectable concentration was 0.01 μg/ml.The recovery rate was 92.6％～97.1％,with a relative standard deviation (RSD) of 3.3％～5.8％.The intra-day and inter-day RSDs were 3.8％～6.2％ and 4.7％～6.3％ respectively.Conclusion This determination method has no requirement for organic solvents,features simple and rapid operation,possesses higher detection sensitivity,and applies well to the determination of 2,5-hexanedione in urine.

Objective To establish a method for determination of 2,5-hexanedione in urine by headspace solid-phase microextraction-gas chromatography.Methods After extraction by solid-phase microextraction head,2,5-hexanedione in urine was determined by gas chromatography and was quantified by external standard method.Results The concentration of 2,5-hexanedione in urine showed a linear relationship within the range of 0.1-20.0 μg/ml.The regression equation was y=261.36x-1.903 3,r=0.999 2.The minimum detectable concentration was 0.01 μg/ml.The recovery rate was 92.6％～97.1％,with a relative standard deviation (RSD) of 3.3％～5.8％.The intra-day and inter-day RSDs were 3.8％～6.2％ and 4.7％～6.3％ respectively.Conclusion This determination method has no requirement for organic solvents,features simple and rapid operation,possesses higher detection sensitivity,and applies well to the determination of 2,5-hexanedione in urine.

Objective:To analyze the effect of nucleos(t)ide analog (NAs) antiviral treatment on the clinical features and prognosis of patients with hepatitis B virus-related hepatocellular carcinoma (HBV-HCC).Methods:A retrospective analysis was performed on the data of 450 HBV-HCC patients first diagnosed and treated in the Third Hospital of Hebei Medical University from January 2015 to January 2021, including 193 patients in the continuous NAs treatment group and 257 patients in the NAs treatment after hepatocellular carcinoma (HCC) group. The baseline data of the two groups were balanced by propensity score matching. The relapse-free survival rate of HCC was estimated by Kaplan-Meier method, and the risk factors for HCC recurrence were analyzed by Cox proportional risk models. Spearman correlation analysis was used to explore the association between clinical features of HCC and hepatitis B virus (HBV) DNA load in patients receiving continuous NAs treatment.Results:Before matching, the proportions of liver cirrhosis, body mass index≥25.0 kg/m 2, single tumor, maximum tumor diameter ≤5 cm, Child-Pugh grade A, China liver cancer staging Ⅰ in the continuous NAs treatment group were 93.8%(181/193), 45.1%(87/193), 70.5%(136/193), 82.4%(159/193), 74.6%(144/193) and 74.6%(144/193), respectively. All of them were higher than those in the NAs treatment after HCC group (87.5%(225/257), 44.0%(113/257), 61.1%(157/257), 55.3%(142/257), 63.8%(164/257) and 56.0%(144/257), respectively). The proportions of drinking history and portal vein tumor thrombi in the continuous NAs treatment group were 12.4%(24/193) and 3.1%(6/193), respectively, which were lower than 33.9%(87/257) and 10.5%(27/257) in the NAs treatment after HCC group.The differences were all statistically significant ( χ2=4.86, 7.58, 4.27, 36.63, 8.15, 21.05, 27.21 and 8.88, respectively, all P<0.05). After matching, the median relapse-free survival time of the patients in the continuous NAs treatment group and the NAs treatment after HCC group were 388 days and 277 days, respectively. The five-year cumulative relapse-free survival rates were 50.0% and 37.5%, respectively, with statistically significant difference ( χ2=5.30, P=0.021). Multivariate analysis showed that no antiviral therapy before diagnosis of HCC, multiple tumors, maximum tumor diameter ≥5 cm and palliative treatment were independent risk factors for HBV-HCC recurrence (hazard ratio ( HR)=1.509, 1.491, 0.446 and 1.472, respectively, all P<0.05). After matching, the maximum tumor diameter ( r=0.175, P=0.042), incidence of portal vein tumor thrombi ( r=0.210, P=0.014) and recurrence of HBV-HCC ( r=0.178, P=0.038) in the continuous NAs treatment group were positively correlated with HBV DNA load. Conclusions:Early initiation of NAs antiviral treatment can improve the tumor characteristics when the disease progresses to HBV-HCC, and improve the relapse-free survival rate of HBV-HCC patients. No antiviral therapy before diagnosis of HCC, multiple tumors, maximum tumor diameter ≥5 cm and palliative treatment are independent risk factors for HBV-HCC recurrence.

Objective To investigate the role of cluster of differentiation (CD) 36 in the formation of non-alcoholic fatty liver disease (NAFLD) in mice. Methods Twenty speciifc pathogen free healthy male C57BL/6J mice [8 weeks old, average weight:(18.8±2.3)g] were randomly divided into NAFLD group and control group according to the random number table method with 10 mice in each group. Mice in NAFLD group was fed with high-fat diet for 10 weeks, while mice in control group was fed with normal diet for 10 weeks. The mice were sacriifced after the processing, and samples of cardiac blood and liver tissue were collected. The levels of serum alanine transaminase (ALT), total cholesterol (TC) and triglyceride (TG), levels of TC and TG in liver tissue, pathological changes of liver tissue, expression of CD36 protein and content of its messenger ribonucleic acid (mRNA) in liver tissue of 2 groups were observed. Experimental data of 2 groups were compared using t test. Results The level of serum ALT was (49±6)U/L in NAFLD group and (45±7)U/L in control group, where no signiifcant difference was obsersed (t=1.70, P>0.05). The levels of serum TC and TG were (4.42±0.09), (0.45±0.04) mmol/L in NAFLD group and (2.42±0.05), (0.32±0.03)mmol/L in control group respectively. The levels of serum TC and TG in NAFLD group were signiifcantly higher than those in control group (t=21.90, 8.22;P<0.05). The levels of TC and TG in liver tissue were (1.18±0.09), (1.75±0.08)mmol/L in NAFLD group and (0.55±0.06), (1.28±0.06)mmol/L in control group respectively. The levels of TC and TG in liver tissue in NAFLD group were signiifcantly higher than those in control group (t=18.42, 14.86; P<0.05). Obviously fatty degeneration and ballooning degeneration were observed in hepatocytes of NAFLD group, while hepatocytes were normal in morphology and size in control group. The expression of CD36 protein in liver tissue in NAFLD group was signiifcantly stronger than that in control group. The content of CD36 mRNA in liver tissue was 2.75±0.26 in NAFLD group and 1.00±0.08 in control group. The content of CD36 mRNA in liver tissue in NAFLD group was significantly higher than that in control group (t=21.16, P<0.05). Conclusions CD36 may play a role in the formation of NAFLD in mice. To decrease its expression can be a new target for the prophylaxis and treatment of NAFLD.

OBJECTIVE: To establish a method for detecting triphenyl phosphate( TPP) in the workplace air by gas chromatography. METHODS: TPP in the air of workplace was collected with glass fiber filter paper,desorbed with ether,separated by HP-5 gas chromatographic column,and detected by flame photometric detector. RESULTS: This method has good linear range of 12. 50-800. 00 mg/L,with the correlation coefficient of 0. 999 9. The detection limit was 0. 78 mg/L,and the minimum detectable concentration was 0. 09 mg/m3(sample volume was 45 L). Desorption efficiency was 97. 2%-99. 4%; standard recovery rate was 99. 5%-100. 3%. The within-run relative standard deviation( RSD) was 2. 7%-3. 4%and the between-run RSD was 1. 4%-3. 2%. The sampling efficiency was 99. 6%-100. 0%. The samples could be stored at room temperature for at least 14 days. CONCLUSION: The method is simple,accurate and highly sensitive for detecting TPP in workplace air.

Objective:To explore the feasibility of predicting TP53 mutation risk by immunohistochemical staining (IHC) pattern of P53 in Chinese diffuse large B-cell lymphoma (DLBCL) and its correlation with a prognostic difference.Methods:Between January 2021 and December 2021, 51 DLBCL cases at Beijing Boren Hospital were gathered. These cases had both IHC and next-generation sequencing (NGS) results. IHC classified the P53 protein expression pattern into a loss (<1% ) , diffuse (>80% ) , and heterogeneous (1% -80% ) . The sensitivity and specificity of the predicting TP53 mutation by IHC were assessed by comparing the results of the NGS, and the TP53 high mutation risk group included both loss and diffuse expression of P53. From June 2016 to September 2019, Peking University Cancer Hospital collected 131 DLBCL cases with thorough clinicopathological and follow-up data. From their tumor blocks, tissue microarray blocks were made for IHC evaluation of P53 expression pattern, and prognosis effect of P53 studies.Results:Among 51 cases with both IHC and NGS results, 23 cases were classified as TP53 high mutation risk (7 cases loss and 16 cases diffuse) , 22/23 cases were proved with mutated TP53 by NGS. Only 1 of the 28 cases classified as TP53 low mutation risk was proved with mutated TP53 by NGS. IHC had a sensitivity and specificity of 95.7% and 96.4% for predicting TP53 mutation. NGS identified a total of 26 TP53 mutations with a mutation frequency of 61.57% (13.41% -86.25% ) . In the diffuse group, 16 missense mutations and 2 splice mutations were detected; 6 truncating mutations and 1 splice mutation were detected in the loss group; 1 truncating mutation was detected in the heterogeneous group. Multivariate analysis demonstrated that TP53 cases with high mutation risk have impartial adverse significance for the 131 patients included in survival analysis ( HR=2.612, 95% CI 1.145-5.956, P=0.022) . Conclusion:IHC of P53 exhibiting loss (<1% ) or diffuse (>80% ) pattern indicated TP53 high mutation risk, IHC can predict TP53 mutation with high specificity and sensitivity. TP53 high mutation risk is an independent predictor for adverse survival.

Objective:To evaluate the efficacy and safety of oral anisodine hydrobromide tablets in the treatment of nonarteritic anterior ischemic optic neuropathy (NAION).Methods:A multicenter nonrandomized controlled trial was conducted.A total of 282 acute NAION patients (282 eyes) were recruited from 16 hospitals in China from July 2020 to May 2021.Patients were divided into two groups according to treatment methods, which were control group (124 cases, 124 eyes) receiving regular treatment including citicoline sodium plus Ginkgo biloba leaf liquid extract or Ginkgo biloba leaf extract tablets plus mecobalamin, and experimental group (158 cases, 158 eyes) receiving treatment in control group plus oral anisodine hydrobromide tablets 1 mg, twice daily for 2 to 3 months.Best corrected visual acuity (BCVA), visual field index (VFI), peripapillary retinal nerve fiber layer (pRNFL) and radial peripapillary capillary vessel density (RPC) were assessed at 1, 2, 3, and 6 months after enrollment using the standard decimal visual acuity chart, 750i Humphery visual field analyzer, Cirrus HD-OCT 4000/Cirrus HD-OCT 5000, RTVue-XR optical coherence tomography respectively.The primary outcomes were BCVA and VFI, and the secondary outcomes were pRNFL, RPC, and the side effects during the follow-up.The study adhered to the Declaration of Helsinki.All patients were fully informed about the treatment and purpose of this study and voluntarily signed the informed consent form.The study protocol was approved by Chinese PLA General Hospital (No.S2020-021-01). Results:In all, 242 patients (242 eyes) completed the follow-up of BCVA, and 98 patients (98 eyes) completed the VFI follow-up.In terms of visual function, BCVA and VFI improved significantly over time in the two groups, and BCVA and VFI were better in experimental group than in control group at various follow-up time points (all at P<0.05). In terms of structure, pRNFL gradually decreased in both groups with the extension of treatment, and pRNFL was significanthy thinner in experimental group than in control group at various follow-up time points (all at P<0.05). There was no significant difference in RPC between the two groups at the last follow-up ( P>0.05). There were two cases with side effects and one case was discontinued due to side effects 25 days after enrollment. Conclusions:Oral anisodine hydrobromide can improve visual acuity and visual field in NAION and accelerate the regression of optic disc edema, with good safety.