AIM To optimize the cellulase-assisted ultrasound extraction process for total flavonoids from Plumbago zeylanica L.,and to evaluate their anti-oxidant activity.METHODS With extraction time,liquid-solid ratio,cellulase addition amount,extraction temperature and ultrasonic power as influencing factors,extraction rate of total flavonoids as an evaluation index,the extraction process was optimized by response surface method on the basis of single factor test.Subsequently,The scavenging rates of extract on DPPH,ABTS and OH free radicals were determined.RESULTS The optimal conditions were determined to be 34∶1 for liquid-solid ratio,3％for cellulase addition amount,51 ℃ for extraction temperature,38 min for extraction time,and 400 W for ultrasonic power,the extraction rate of total flavonoids was(33.411±0.97)％.The IC50 values of three free radicals were 0.13,0.042,3.29 mg/mL,respectively.CONCLUSION This reasonable and reliable method can be used for the cellulase-assisted ultrasound extraction of total flavonoids from P.zeylanica with strong anti-oxidant activity.

AIM To establish an HPLC-MS/MS method for the simultaneous content determination of gallic acid,protocatechuic acid,oxypaeoniflorin,catechin,epicatechin,albiflorin,paeoniflorin,rutin,calycosin-7-glucoside,syringaldehyde,ferulic acid,coumarin,ononin,calycosin,cinnamic alcohol,cinnamic acid,benzoylpaeoniflorin,cinnamaldehyde,astragaloside,astragaloside Ⅲ,6-gingerol in Huangqi Guizhi Wuwu Decoction.METHODS The analysis was performed on a 30 ℃ thermostatic Thermo Scientific Hypersil GOLD column(150 mmx4.6 mm,3 μm),with the mobile phase comprising of 0.015％formic acid-acetonitrile flowing at 0.4 mL/min in a gradient elution manner,and electrospray ionization source was adopted in positive and negative ion modes with multiple reaction monitoring.RESULTS Twenty-one constituents showed good linear relationships within their own ranges(r＞0.990 5),whose average recoveries were 93.99％-108.52％with the RSDs of 1.04％-5.97％.CONCLUSION This simple,feasible,stable and reliable method can be used for the quality control of Huangqi Guizhi Wuwu Decoction.

AIM To establish an HPLC-MS/MS method for the simultaneous content determination of gallic acid,protocatechuic acid,oxypaeoniflorin,catechin,epicatechin,albiflorin,paeoniflorin,rutin,calycosin-7-glucoside,syringaldehyde,ferulic acid,coumarin,ononin,calycosin,cinnamic alcohol,cinnamic acid,benzoylpaeoniflorin,cinnamaldehyde,astragaloside,astragaloside Ⅲ,6-gingerol in Huangqi Guizhi Wuwu Decoction.METHODS The analysis was performed on a 30 ℃ thermostatic Thermo Scientific Hypersil GOLD column(150 mmx4.6 mm,3 μm),with the mobile phase comprising of 0.015％formic acid-acetonitrile flowing at 0.4 mL/min in a gradient elution manner,and electrospray ionization source was adopted in positive and negative ion modes with multiple reaction monitoring.RESULTS Twenty-one constituents showed good linear relationships within their own ranges(r＞0.990 5),whose average recoveries were 93.99％-108.52％with the RSDs of 1.04％-5.97％.CONCLUSION This simple,feasible,stable and reliable method can be used for the quality control of Huangqi Guizhi Wuwu Decoction.

AIM To optimize the cellulase-assisted ultrasound extraction process for total flavonoids from Plumbago zeylanica L.,and to evaluate their anti-oxidant activity.METHODS With extraction time,liquid-solid ratio,cellulase addition amount,extraction temperature and ultrasonic power as influencing factors,extraction rate of total flavonoids as an evaluation index,the extraction process was optimized by response surface method on the basis of single factor test.Subsequently,The scavenging rates of extract on DPPH,ABTS and OH free radicals were determined.RESULTS The optimal conditions were determined to be 34∶1 for liquid-solid ratio,3％for cellulase addition amount,51 ℃ for extraction temperature,38 min for extraction time,and 400 W for ultrasonic power,the extraction rate of total flavonoids was(33.411±0.97)％.The IC50 values of three free radicals were 0.13,0.042,3.29 mg/mL,respectively.CONCLUSION This reasonable and reliable method can be used for the cellulase-assisted ultrasound extraction of total flavonoids from P.zeylanica with strong anti-oxidant activity.

Objective To explore the mechanism of somatostatin in improving acute lung injury associated with acute pancreatitis.Methods Wistar rats were randomly divided into sham operation group(injection of normal saline),model group(puncture of common bile duct and injection of 5％sodium taurocholate with wire ligation),somatostatin group(injection of somatostatin into tail vein of model group),somatostatin+miR-146a-5p inhibitor group(on the basis of somatostatin group,tail vein injection of miR-146a-5p inhibitor and somatostatin+oe-angiogenin-like protein 4(ANGPTL4)group(on the basis of somatostatin group,tail vein injection of oe-ANGPTL4 plasmid).Hematoxylin-eosin(HE)staining was used to observe the pathological changes of pancreatic and lung tissues;pathological score and tissue wet-dry weight ratio were determined,real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)was used to detect miR-146a-5p and ANGPTL4 mRNA expression and Western blot was used to detect the expression of related proteins in lung tissues of rats.Tumor necrosis factor-α(TNF-α)was detected by enzyme-linked immunosorbent assay(ELISA).Results In sham operation group,model group and somatostatin group,the damage degree of pancreas tissue(based on modified computed tomography severity index)were 1.25±0.28,3.20±0.34,2.15±0.31,respectively;the damage degree of lung tissue(based on the Smith lung injury score system)were 1.40±0.13,5.10±0.58,3.10±0.38,respectively.The relative expression levels of ANGPTL4 mRNA in sham operation group,model group,somatostatin group and somatostatin+miR-146a-5p inhibitor group were 1.00±0.17,1.63±0.20,1.21±0.18 and 1.73±0.28.The levels of TNF-α in sham operation group,model group,somatostatin group,somatostatin+miR-146a-5p inhibitor group and somatostatin+oe-ANGPTL4 group were(76.33±7.25),(125.05±13.56),(80.11±10.68),(118.62±14.32)and(105.32±13.52)pg·mL-1,respectively;the relative expression levels of nuclear factor E2-related factor 2(Nrf2)protein were 1.00±0.27,0.51±0.07,0.88±0.14,0.68±0.12,0.51±0.09,respectively;the relative expression levels of heme oxygenase-1(HO-1)protein were 1.00±0.25,0.58±0.11,0.79±0.18,0.48±0.07 and 0.50±0.08,respectively.The above indexes of the model group were compared with those of the sham operation group,and the above indexes of the somatostatin group were compared with those of the model group,somatostatin+miR-146a-5p inhibitor group and somatostatin+oe-ANGPTL4 group,and the differences were statistically significant(all P＜0.05).Conclusion Somatostatin has antioxidant and anti-inflammatory effects and can ameliorate acute lung injury associated with acute pancreatitis.The mechanism may be related to Nrf2/HO-1 pathway mediated by miR-146a-5p/ANGPTL4.

Aim To explore the mechanism of the effect of rosiglitazone(Rsg)on the pancreatic cancer in diabetic mice based on the impact of PPARγ on glu-cose transport and metabolism.Methods A high-fat and high sugar diet combined with STZ was used to construct T2DM model;T2DM mice and normal mice were subcutaneously injected with PANC02 cells to construct a transplanted tumor model.T2DM trans-planted tumor mice and normal transplanted tumor mice were divided into the following groups:Rsg,PPARy inhibitor(PIN-2),rosiglitazone+PPARγ in-hibitor(Rsg+PIN-2),and normal transplanted tumor mice(NDM)and T2DM transplanted tumor mice(DM)were used as control groups,respectively.Tis-sue samples were collected after intervention.Tissue pathological changes were observed by HE staining.The expressions of Ki67 and PCNA proteins were de-tected by immunohistochemistry.Cell apoptosis was detected by TUNEL assay.The expression of PPARγwas detected by immunofluorescence.The expressions of Glucokinase,GLUT2,Nkx6.1,PDX-1RT-PCR were determined by Western blot.Results Rsg could significantly reduce the tumor mass,pathological chan-ges,Ki67 and PCNA expression of transplanted tumors(P＜0.05),increase cell apoptosis and the expression of PPARγ,Glucokinase,GLUT2,Nkx6.1,PDX-1 proteins in NDM and DM mice(P＜0.05).PIN-2 could reverse the indicator changes caused by Rsg in NDM and DM mice.However,compared with NDM mice,the above related indicators of the DM group mice were more sensitive to Rsg and PIN-2.Conclu-sions Compared to non-diabetic pancreatic cancer,rosiglitazone can more sensitively inhibit the prolifera-tion of pancreatic cancer with T2DM,induce apopto-sis,and reprogram the metabolism of pancreatic cancer with T2DM by activating PPA Rγ and altering the ex-pression of glucose and lipid metabolism genes,there-by exerting an anti-cancer effect.

OBJECTIVE: Clinical characteristics and outcome in COVID-19 with brucellosis patients has not been well demonstrated, we tried to analyze clinical outcome in local and literature COVID-19 cases with brucellosis before and after recovery. METHODS: We retrospectively collected hospitalization data of comorbid patients and prospectively followed up after discharge in Heilongjiang Infectious Disease Hospital from January 15, 2020 to April 29, 2022. Demographics, epidemiological, clinical symptoms, radiological and laboratory data, treatment medicines and outcomes, and follow up were analyzed, and findings of a systematic review were demonstrated. RESULTS: A total of four COVID-19 with brucellosis patients were included. One patient had active brucellosis before covid and 3 patients had nonactive brucellosis before brucellosis. The median age was 54.5 years, and all were males (100.0%). Two cases (50.0%) were moderate, and one was mild and asymptomatic, respectively. Three cases (75.0%) had at least one comorbidity (brucellosis excluded). All 4 patients were found in COVID-19 nucleic acid screening. Case C and D had only headache and fever on admission, respectively. Four cases were treated with Traditional Chinese medicine, western medicines for three cases, no adverse reaction occurred during hospitalization. All patients were cured and discharged. Moreover, one case (25.0%) had still active brucellosis without re-positive COVID-19, and other three cases (75.0%) have no symptoms of discomfort except one case fell fatigue and anxious during the follow-up period after recovery. Conducting the literature review, two similar cases have been reported in two case reports, and were both recovered, whereas, no data of follow up after recovery. CONCLUSION These cases indicate that COVID-19 patients with brucellosis had favorable outcome before and after recovery. More clinical studies should be conducted to confirm our findings.
Female ; Humans ; Male ; Middle Aged ; Brucellosis ; COVID-19 ; Retrospective Studies ; SARS-CoV-2 ; Treatment Outcome ; Case Reports as Topic

Objective: To understand the knowledge, attitude, and current status of vaccination of herpes zoster vaccination among urban residents aged 25 years and above in China. Methods: In August to October 2022, a convenience sampling method was used to survey residents aged 25 years and above at 36 community centers in 9 cities across China. Questionnaires were used to collect basic information, knowledge, and attitude toward herpes zoster and its vaccination, as well as vaccination status and reasons for non-vaccination among residents. Results: A total of 2 864 urban residents were included in the study. The total score of residents' cognition of herpes zoster and its vaccine was 3.01±2.08, and the total score of their attitude was 18.25±2.76. Factors such as being male (β=-0.45, P<0.001), older than 40-59 years (β=-0.34, P=0.023) or ≥60 years (β=-0.68, P<0.001), married (β=-0.69, P=0.002) were negatively associated with knowledge score. The educational level of high school or secondary school (β=0.44, P=0.036), college (β=0.65, P=0.006), bachelor's degree and above (β=1.20, P<0.001), annual net household income ≥120 000 Yuan in 2021 (β=0.42, P=0.020), having urban employee medical insurance (β=0.62, P=0.030), having public or commercial medical insurance (β=0.65, P=0.033), and having a history of chickenpox (β=0.29, P=0.025) were positively associated with knowledge scores. Being male (β=-0.38, P=0.008) and not remembering a history of chickenpox (β=-0.49, P=0.012) were negatively associated with attitude scores. Annual net household income in 2021 was between 40 000-80 000 Yuan (β=0.44, P=0.032) or between 80 000-120 000 Yuan (β=0.62, P=0.002) or ≥120 000 Yuan (β=0.93, P<0.001), and a history of herpes zoster (β=0.59, P=0.004) were positively associated with attitude scores. Of the 2 864 residents surveyed, only 29 (1.01%) had received the herpes zoster vaccine, with a vaccination rate of 1.70% for those aged 50 years and above, with the main reason for non-vaccination being lack of knowledge about the herpes zoster vaccine, followed by the high price. 42.67% of the population said they would consider getting the herpes zoster vaccine in the future. Conclusion: Low knowledge of herpes zoster and its vaccine, positive attitudes towards the preventive effects of herpes zoster and its vaccine, and extremely low vaccination rates among the urban population in China call for multiple measures to strengthen health education and vaccination recommendations for residents, especially for the elderly, low-education and low-income populations.
Aged ; Male ; Humans ; Female ; Herpes Zoster Vaccine ; Chickenpox ; Health Knowledge, Attitudes, Practice ; Urban Population ; Herpes Zoster/prevention & control* ; China

Objective: To investigate the influence of HBsAg expression in peritumoral tissue of hepatocellular carcinoma (HCC) patients on their postoperative recurrence. Methods: The HCC patients treated in Shanghai Eastern Hepatobiliary Surgery Hospital from October 2009 to August 2010 were selected. The clinicopathological data and adjacent tissues of 718 patients were collected, and dextran polymer immunohistochemical staining was used to detect the expression of HBsAg in adjacent tissues. According to the expression of HBsAg in adjacent tissues, the tissues were divided into HBsAg positive group and HBsAg negative group. Kaplan-Meier method and Log rank test were used for survival analysis, and Cox regression model was used for multivariate analysis. Results: Among the 718 patients in the whole group, 153 were HBsAg negative and 565 were HBsAg positive. There was a statistically significant difference in serum HBV DNA level between HBsAg-positive and HBsAg-negative patients (P<0.001). The number of patients with serum DNA≥2 000 IU/ml and<2 000 IU/ml in HBsAg negative group were 52 and 93, while the patients in HBsAg positive group were 325 and 205. The cumulative recurrence rates of all patients at 1, 3, and 5 years after surgery were 30.2%, 54.3%, and 62.7%, respectively. The expression of HBsAg was related to the recurrence (P=0.038). Multivariate analysis showed that γ-GT, PT, multiple tumors, tumor length, and portal vein invasion were independent risk factors for recurrence of HCC (P<0.05). In HBeAg-negative patients with low viral load (HBV DNA <2 000 IU/ml) and without cirrhosis, the recurrence rates of HBsAg-positive patients were 14.3% and 31.0% at 3 and 5 years, respectively, compared with HBsAg negative patients (all 0), the difference was statistically significant (P=0.021). Conclusion: The positive expression of HBsAg in peritumoral tissue increases the postoperative recurrence risk of HCC patients.
Carcinoma, Hepatocellular/pathology* ; China ; DNA, Viral/analysis* ; Hepatitis B Surface Antigens ; Hepatitis B virus/metabolism* ; Humans ; Liver Neoplasms/pathology*

Obvious epigenetic differentiation occurred on Lycium barbarum in different cultivation areas in China. To investigate the difference and change rule of DNA methylation level and pattern of L. barbarum from different cultivation areas in China, the present study employed fluorescence-assisted methylation-sensitive amplified polymorphism(MSAP) to analyze the methylation level and polymorphism of 53 genomic DNA samples from Yinchuan Plain in Ningxia, Bayannur city in Inner Mongolia, Jingyuan county and Yumen city in Gansu, Delingha city in Qinghai, and Jinghe county in Xinjiang. The MSAP technical system suitable for the methylation analysis of L. barbarum genomic DNA was established and ten pairs of selective primers were selected. Among amplified 5'-CCGG-3' methylated sites, there were 35.85% full-methylated sites and 39.88% hemi-methylated sites, showing a high degree of epigenetic differentiation. Stoichiometric analysis showed that the ecological environment was the main factor affecting the epigenetic characteristics of L. barbarum, followed by cultivated varieties. Precipitation, air temperature, and soil pH were the main ecological factors affecting DNA methylation in different areas. This study provided a theoretical basis for the analysis of the epigenetic mechanism of L. barbarum to adapt to the diffe-rent ecological environments and research ideas for the introduction, cultivation, and germplasm traceability of L. barbarum.
China ; DNA Methylation ; DNA Primers ; Epigenesis, Genetic ; Lycium/genetics*