Objective:To prepare 7-hydroxyethyl chrysin(7-HEC)loaded poly(lactic-co-glycolic acid)(PLGA)nanoparticles and to detect the in vitro release.Methods:The 7-HEC/PLGA nanoparticles were prepared by emulsification solvent volatilization method.The particle size,polydispersity index(PDI),encapsulation rate,drug loading and zeta potential were measured.The prescription was optimized by single factor investigation combined with Box-Behnken response surface method.Mannitol was used as protectant to prepare lyophilized powder,and the optimal formulation was characterized and studied for the in vitro release.Results:The optimal formulation of 7-HEC/PLGA nanoparticles was as follows:drug loading ratio of 2.12∶20,oil-water volume ratio of 1∶14.7,and 2.72%soybean phospholipid as emulsifier.With the optimal formulation,the average particle size of 7-HEC/PLGA nanoparticles was(240.28±0.96)nm,the PDI was 0.25±0.69,the encapsulation rate was(75.74±0.80)%,the drug loading capacity was(6.98±0.83)%,and the potentiostatic potential was(-18.17±0.17)mV.The cumulative in vitro release reached more than 50%within 48 h.Conclusions:The optimized formulation is stable and easy to operate.The prepared 7-HEC/PLGA nanoparticles have uniform particle size,high encapsulation rate and significantly higher dissolution rate than 7-HEC.

Objective:To explore the mechanism by which Pien Tze Huang improves liver Kupffer cell damage induced by lipopolysaccharide (LPS) by regulating the expression of miR-155.Methods:LPS induced liver Kupffer cells to establish a cell injury model to simulate septic liver injury. RT-qPCR was used to detect the expression of miR-155 in damaged cells, and RT-qPCR, Western Blot, ELISA and flow cytometry were used to evaluate the inflammatory response and apoptosis of damaged cells. Then we treated LPS-induced Kupffer cells with Pien Tze Huang at different concentrations (0 mg/L, 5 mg/L, 10 mg/L and 15 mg/L), and detected the expression of miR-155 in the cells, the inflammatory response of the cells and Apoptosis rate. MiR-155 was silenced in the cell injury model, and RT-qPCR, Western Blot, ELISA and flow cytometry were used to evaluate the effect of miR-155 on inflammatory response and apoptosis of model cells. Overexpression of miR-155 in damaged cells treated with Pien Tze Huang was used to detect changes in cellular inflammatory response and apoptosis. Data are expressed in the form of mean ± standard deviation, and each group of data is analyzed using t test or one-way analysis of variance.Results:In the LPS-induced liver Kupffer cell injury model, the expression of miR-155 was significantly increased ( P<0.05), the expression levels of pro-inflammatory factors IL-6 and TNF-α were significantly increased, and the anti-inflammatory factor IL-10 was significantly increased. was inhibited ( P<0.05), and the cell apoptosis rate was significantly increased ( P<0.05). After Pien Tze Huang treatment, the expression of miR-155 in damaged liver cells was inhibited ( P<0.05), the levels of cellular inflammatory factors IL-6 and TNF-α were inhibited, and the expression of anti-inflammatory factor IL-10 was promoted ( P<0.05). Inhibit cell apoptosis ( P<0.05). Silencing miR-155 reduced the inflammatory response and apoptosis rate of cells ( P<0.05). Overexpression of miR-155 can reverse the effect of Pien Tze Huang on liver cell injury ( P<0.05). Conclusions:In the model of LPS-induced liver Kupffer cell injury, Pien Tze Huang can inhibite the inflammatory response and apoptosis of cells by inhibiting the expression of miR-155.

Objective:To investigate the changes of exam scores of retrained peritoneal dialysis (PD) operators (patients, family members, or nannies) with an internal of one year during COVID-19 epidemic and provide basis for targeted training.Methods:It was a cross-sectional survey study. The maintenance PD patients who participated in two trainings with an interval of one year during COVID-19 epidemic from November 1, 2019 to February 28, 2021 in Department of Nephrology in Peking University People's Hospital were enrolled. During COVID-19 epidemic, retraining was extended from once every six months to once a year. The clinical data were collected, the self-designed training exam score table including theoretical knowledge and operational skills assessment was used to investigate the exam scores of two trainings, and the total exam scores and sub-item scores of PD operators before and after one year were compared. Logistic regression analysis was used to analyze the associated factors of the reduction of exam scores.Results:A total of 59 patients were enrolled, with 35 males (59.32%), age of (58.41±14.52) years, and dialysis duration of 42 (12, 84) months. There were 54 patients (91.53%) operating by themselves, 22 operators (37.29%) having college degree or above, and 35 operators (59.32%) having decreased exam scores. The total exam scores were 83.17±7.90 and 80.61±8.20 before and after one year, respectively ( t=2.732, P=0.008). In the six contents of itemized scoring, compared with one year ago, the exam scores of complication treatment ( t=4.928, P<0.001) and self-monitoring ( t=3.222, P=0.002) were significantly decreased. There was no statistically significant difference in the exam scores of environment and hygiene, dialysate replacement operation, exit nursing and diet before and after one year (all P>0.05). The total exam scores in patients with dialysis duration <12 months and 36-60 months after one year were significantly lower than before one year ( t=2.309, P=0.041; t=3.086, P=0.009). There was no statistically significant difference in the exam scores of PD operators with dialysis duration of 12-<36 months and >60 months before and after one year (both P>0.05). Logistic regression analysis showed that dialysis duration was an independent associated factor of exam scores reduction (dialysis duration 36-60 months/>60 months, OR=6.233, 95% CI 1.035-37.529, P=0.046). Conclusions:During COVID-19 epidemic, the reduced frequency of retraining reduces the training exam scores of PD operators, especially in patients with dialysis duration of 36-60 months. The weak points are focused on complication management and self-monitoring. Training should be strengthened for key patients and key contents if regular retraining is not possible due to special circumstances.

Objective:To analyze the risk factors of carbapenem-resistant Klebsiella pneumoniae（CRKP）infection in tumor patients. Methods:The clinical data of hospitalized tumor patients with Klebsiella pneumoniae（KP）infection admitted in Hangzhou Cancer Hospital during January 2021 to December 2022 were retrospectively analyzed. Patients were divided into CRKP group（ n=37）and carbapenem-sensitive Klebsiella pneumoniae（CSKP）group（ n=142）according to drug sensitivity. The risk factors of CRKP infection were analyzed by univariate and multivariate Logistic regression. Results:Univariate analysis showed that cardiovascular disease（ χ2=4.554， P=0.033），carbapenem use ≥7 days（ χ2=53.483， P<0.001）and hospital stay >7 days（ χ2=3.871， P=0.049）were significantly associated with CRKP infection in tumor patients. Multivariate Logistic analysis showed that carbapenem therapy for ≥7 days（ OR=18.280，95% CI 6.533-51.147， P<0.001）was an independent risk factor for CRKP infection in tumor patients. Conclusion:Tumor patients with long-term use of carbapenems are prone to CRKP infection.

Objective To observe the ultrasonic manifestations and outcomes of fetal congenital kidney anomalies(CKA)during the second trimester of pregnancy.Methods Data of 223 singleton fetuses with CKA detected with ultrasound during the second trimester of pregnancy were retrospectively analyzed.The ultrasonic manifestations of fetal CKA during the second trimester of pregnancy were observed,and the changes till the third trimester of pregnancy were analyzed.Results Among 223 CKA fetuses,simple renal pelvis separation was detected in 142 fetuses,while simple double renal pelvis malformation in was found in 41,polycystic kidney disease in 11,absent kidneys in 7,simple renal cysts and ectopic kidneys each in 6,smaller kidneys in 4,horseshoe kidneys and unilateral renal pelvis separation complicated with contralateral double renal pelvis malformation each in 2,unilateral renal cyst swelling complicated with contralateral double renal pelvis malformation and unilateral renal double renal pelvis malformation complicated with renal pelvis separation was noticed each in 1 fetus.Abnormalities such as renal pelvis separation,renal pelvis malformation,renal cysts and smaller kidney changed during the third trimester of pregnancy,while polycystic kidney disease,absent kidney,ectopic kidney and horseshoe kidney not significantly changed.Conclusion Fetal CKA had various types being able to change dynamically with the development of pregnancy,among which renal pelvis separation was the most common type.

Objective To observe the ultrasonic manifestations and genetic abnormalities in the second trimester of pregnancy of fetal congenital kidney anomalies(CKA).Methods Totally 14 singleton pregnancy women with CKA fetus detected with prenatal ultrasound and proved by genetic examination who underwent prenatal ultrasound in the second trimester of pregnancy were enrolled.The ultrasonic manifestations and genetic abnormalities in the second trimester of pregnancy were observed.Results Among 14 fetuses,simple renal abnormalities were detected in 10,while renal abnormalities complicated with extrarenal abnormalities were noticed in 4 fetuses with prenatal ultrasound in the second trimester of pregnancy,presented as enhanced renal echoes,renal pelvis separation and ectopic kidney,etc.Genetic examination showed chromosomal karyotype abnormalities in 4 fetuses,chromosomal copy number abnormalities in 8 fetuses,and genetic mutations in 2 fetuses.Conclusion The ultrasonic manifestations in the second trimester of pregnancy of fetal CKA included enhanced renal echo,renal pelvis separation and ectopic kidney,etc.Severe chromosomal or genetic abnormalities could exist even when the relative manifestations were not obvious.

Objective:To explore independent predictors for poor outcome at 3 months in elderly patients with acute cerebral infarction(ACI)treated with intravenous thrombolysis(IVT), and to develop a nomogram-based predictive model.Methods:This was a retrospective cohort study.Clinical, laboratory and imaging data of 346 elderly patients with ACI treated with IVT from January 2016 to April 2021 in our hospital were collected.Poor outcome was defined as a modified Rankin Scale(mRS)score >2 at 3 months after the stroke.Logistic regression analysis was used to screen for independent factors predicting poor outcome in elderly ACI patients treated with IVT, and a corresponding nomogram model was developed using the R software.The ROC curve, calibration plots and decision curve analysis were used to evaluate discrimination, calibration and clinical application value of the nomogram model.Results:Among 346 candidates, 109 developed a poor outcome, representing a rate of 31.5%.Logistic regression analysis showed that symptomatic hemorrhagic transformation( OR=15.647, 95% CI: 8.913-27.454), stroke severity(moderate stroke, OR=3.322, 95% CI: 1.414-7.811; moderate-severe stroke, OR=8.169, 95% CI: 4.102-16.258; severe stroke, OR=9.653, 95% CI: 5.440-17.121), stroke-associated pneumonia( OR=2.239, 95% CI: 1.134-4.420), and heart failure( OR=2.758, 95% CI: 1.424-5.336)were independent predictors for poor outcome at 3 months in elderly ACI patients treated with intravenous thrombolysis(all P<0.05). With the area under curve(AUC-ROC)value at 0.85(95% CI: 0.80-0.89), the nomogram model, which was composed of the above four predictors, demonstrated good discrimination.On the calibration plot, the mean absolute error was 0.020, indicating that the model had good calibration.Decision curve analysis revealed that the model had good clinical application value. Conclusions:The nomogram model composed of symptomatic hemorrhagic transformation, stroke severity, stroke-associated pneumonia and heart failure may predict poor outcome at 3 months in elderly ACI patients treated with IVT, with high prediction accuracy and high clinical application value.

Objective To construct SLC6A4-shRNA lentiviral vector, establish PC12 cells stable transformation cell line，and detect the effect of SLC6A4 gene silencing on hypoxia induced PC12 cells apoptosis. Methods Three specific targets sequence of SLC6A4 were designed and short hairpin RNA was synthesized, and then were recombined into shRNA expression vector GV248 plasmid, with non-homology shRNA sequence as negative control. The connection products were switched to competent cells. After dentification and sequencing, the vectors were co-transfected with the auxiliary vectors into 293T cells in order to produce recombinant shRNA lentiviral particles. Then, PC12 cells were infected with the recombinant lentiviral and screened by puromycin. The PC12 cells were divided into two groups: lentiviral negative control group (NC-shRNA) and SLC6A4-silenced group (SLC6A4-shRNA). The expression of SLC6A4 mRNA was detected by real-time fluorescence quantitative and the 5-HTT protein level was assayed by Western blot, The effect of SLC6A4 gene silencing on hypoxia induced apoptosis was detected by flow cytometry. Results The SLC6A4-shRNA lentiviral expression vector was constructed and the recombinant lentiviral particles by packaging the 293T cells were obtained, the stably infected PC12 cells were established after filtering. Compared with negative control group, the expression level of SLC6A4 gene and 5-HTT protein in SLC6A4-shRNA group was suppressed notably (P<0.01). It was confirmed that lentiviral vector could effectively silence SLC6A4 gene in PC12 cells and SLC6A4 gene silencing could decrease apoptosis rate of PC12 cells under hypoxia condition. Conclusion The SLC6A4 gene of PC12 cells could be effectively silenced by shRNA lentivirus vector，which could reverse hypoxia induced apoptosis.

Objective To study the protective effect and mechanism of Saussurea involucrata flavone capsule on the myocardial tissue of mice in simulated plateau hypoxia environment. Methods 64 Balb/c mice were randomly divided into normal control group, hypoxia model group, positive control group and Xuelian flavonoid capsule group. Myocardial tissue was observed microscopically and ultra-structurally, and the changes of hypoxia-related indexes were detected. The changes in the transcription levels of hypoxia-related genes were detected by RT-PCR, and the changes in the protein expressions of hypoxia-related genes were detected by Western blotting to study the mechanism of action of Xuelian flavone capsules. Results Saussurea involucrata flavone capsule had significantly alleviated the pathological damage to the myocardium of mice caused by simulated altitude hypoxia, increased the activity of T-AOC in myocardial tissue, reduced the accumulation of LD, and also reduced the expression levels of HIF-1α and VEGF mRNA in myocardial tissue, increased the SOD, CAT mRNA and protein expression levels. Conclusion Saussurea involucrata flavone capsule have good anti-altitude hypoxia effect, which could protect myocardial tissue structure and function, regulate energy metabolism, and improve antioxidant capacity. The mechanism might be related to improving the antioxidant capacity, regulating energy metabolism, and affecting the protein expression of HIF-1α, VEGF, SOD and CAT mRNA in high altitude hypoxic mice.

ObjectiveTo analyze the epidemic characteristics of foodborne disease outbreaks in Mengla County， Yunnan Province， so as to provide basis for formulating corresponding prevention and control measures. MethodsThe data of foodborne disease outbreaks in Mengla County， Yunnan Province from 2017 to 2021 in the national "foodborne disease outbreak monitoring system" were collected. The time， population， region， place， pathogenic factors and inducing links of the events were statistically analyzed. ResultsFrom 2017 to 2021， a total of 68 incidents were reported， including 526 cases and 5 deaths. The total incidence rate was 23.40% and the case fatality rate was 0.95%. The peak period was from May to July， with the largest number of reported events in July. The reporting areas were mainly Mengla Town （35.29%）， Guanlei Town （11.76%） and Mengpeng Town （10.29%）. The main place of the incident was family （76.47%）. The main pathogenic factors were plant toxins （79.41%）， mainly through the ingestion of wild mushrooms （68.52%）， improper processing of green beans （12.96%） and aconitum （9.26%）. ConclusionAccording to the epidemiological characteristics of Mengla County， we should focus on the public education of health knowledge in key areas and places， especially on the identification of wild mushrooms， to prevent accidental eating. At the same time， we should strengthen the monitoring and risk assessment of foodborne diseases， find potential risks as soon as possible， issue early warning and forecast in time， and constantly improve the clinical treatment ability of poisoned patients.