Objective To observe the synthesis of melanin and tyrosinase activity of the cultured normal human melanocytes after treated with baicalin and NB-UVB, and to provide theoretical evidence for study on the new therapy method of vitiligo.Methods The normal human menlanocytes cultured in vitro were divided into control group,different concentrations of baicalin groups,different doses of NB-UVB group and baicalin combined NB-UVB group.The proliferation rate of melanocytes was measured by MTT assay,the level of melanin was tested by NaOH pyrolysis,and the tyrosinase activity was measured by dopa oxidization.Results Compared with control group,the proliferation rates in 10-4 mol·L-1 baicalin group and 30 mJ·cm-2 NB-UVB group had no significant difference(P>0.05 ). Compared with control group, the levels of melanin and tyrosinase activities in baicalin group,NB-UVB group and combined group were increased (P<0.05 or P<0.01);compared with baicalin group and NB-UVB group,the level of melanin and tyrosinase activity in combined group were increased(P<0.01). Conclusion Baicalin combined with NB-UVB has better promotion effect on melanin synthesis than used alone.

OBJECTIVE:To improve the quality standard of Jianpi zhixiening granules. METHODS:TLC was applied for qual-itative identification of Scutellaria baicalensis,Crategi Fructus,Lonicerae japonicae,Codonopsis Radix,Nelumbo nucifera,Copti-dis Rhizoma. The contents of berberine hydrochloride and baicalin were determined by HPLC. The determination was performed on Shimadzu VP-ODS column with mobile phase consisted of methanol-acetonitrile-water (46:30:42,V/V/V,berberine hydrochlo-ride))(0.1% sodium dodecylsulphate,0.1% phosphoric acid,methanol-0.4% phosphoric acid(50:50,V/V,baicalin)at the flow rate of 1.0 mL/min. The detection wavelengths were set at 265 nm (berberine hydrochloride) and 280 nm (baicalin). The column temperature was 30 ℃,and sample size was 10 μL. RESULTS:TLC spots of S. baicalensis,Crategi Fructus,L. Japonicae,Co-donopsis Radix,N. nucifera,Coptidis Rhizoma were clear and well-separated without negative interference. The linear ranges of berberine hydrochloride and baicalin were 6.67-33.34 μg/mL(r=0.9998)and 7.7-38.7 μg/mL(r=0.9999). RSDs of precision,sta-bility and reproducibility tests were all lower than 1.0% . The recoveries were 96.5% -99.9%(RSD=1.2% ,n=6)and 101.1%-102.9%(RSD=0.6%,n=6). CONCLUSIONS:Improved standard can be used for quality control of Jianpi zhixiening granules.

Objective:To explore the clinical significance of combined detection of the promoter methylation of plasma free Septin9, SDC2 and BCAT1 genes in peripheral blood for the diagnosis of colorectal cancer. Methods The data of patients admitted to the Department of Gastroenterology, Shanghai East Hospital Affiliated to Tongji University from January to September 2019 were retrospectively analyzed. They were divided into colorectal cancer group (62 cases of colon cancer, 59 cases of rectal cancer), precancerous lesions group (77 cases of colorectal adenoma, 5 cases of high-grade intraepithelial neoplasia), interference group (61 cases of colorectal cancer and advanced adenoma negative but suffered other intestinal lesions, 17 cases of non-colorectal cancer) and healthy group (94 cases). The methylation status of three genes (Septin9, SDC2 and BCAT1) in peripheral blood plasma was detected simultaneously by fluorescence PCR. The relationship between the positive rate of three genes detected jointly and the clinic pathological characteristics of colorectal cancer was analyzed and compared with serum carcinoembryonic antigen (CEA) positive rate. The colorectal cancer group was divided into stage Ⅰ, Ⅱ, Ⅲ and Ⅳ according to TNM stage, and the colorectal cancer group was analyzed and counted by grade. The diagnostic efficiency of detection methods was analyzed by receiver operating characteristic (ROC) curve, and the area under ROC curve (AUC) was compared.Results:The positive rate of combined detection of SDC2 and BCAT1 gene methylation was higher than other three groups (χ 2 =237.246, P<0.001). The positive rate of combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation was higher than CEA in colorectal cancer group ( P<0.001). The positive rates of the combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation in stage Ⅰ-Ⅳ of colorectal cancer group were 73%(16/22), 87%(34/39), 86%(30/35) and 96%(24/25), respectively. Compared with CEA group, the positive rate of combined detection of plasma Septin9, SDC2 and BCAT1 gene methylation in stage Ⅰ-Ⅲ of colorectal cancer group was higher than serum CEA ( P<0.001), but the positive rate of stage Ⅳ was not statistically significant compared with CEA group ( P>0.05). ROC curve analysis showed that the AUC of Septin9, SDC2 and BCAT1 was 0.857(95% CI 0.810-0.903),0.819(95% CI 0.768-0.871)and 0.862(95% CI 0.816-0.909), respectively. The AUC of combined detection of three gene methylations was 0.889 (95% CI 0.846-0.933), and the AUC of combined detection with serum CEA was 0.913 (95% CI 0.874-0.951). There was no significant difference in the positive rate of combined detection of Septin9, SDC2 and BCAT1 gene methylation among different gender, age and cancerous site of colon cancer patients (all P>0.05). Conclusion:The combined detection of the promoter methylation of plasma free Septin9, SDC2 and BCAT1 genes in peripheral blood plasma is helpful for the early diagnosis of colorectal cancer. The positive rate in stage Ⅰ-Ⅲ of colorectal cancer group is higher than serum CEA. The combined diagnosis of the three genes can improve the diagnostic efficiency.

ObjectiveTo detect the fiber structure and volume differences in frontal lobe between patients with amnestic mild cognitive impairment (aMCI) and normal control.MethodsT1 -weighted magnetic resonance imaging and diffusion tensor imaging were obtained in 28 aMCI patients and 25 normal controls.Volumes,fiber fractional anisotropy (FA),fiber apparent diffusion coefficient (ADC),fiber number,and average fiber length of frontal lobe in the two groups were measured.ResultsVolumes of left frontal lobe ( ( 337.35 ± 20.45 ) cm3 ) in aMCI group were smaller than control ( (358.54 ± 27.26) cm3 ) ( t =- 3.223,P =0.002 ).Numbers of short range fiber in left frontal lobe ( 16985 ± 892) were significantly increased relative to control ( 16387 ±752) ; while numbers of long range fiber (3214 ± 185 ) were reduced compared with control (3425 ± 277 ),and ADC values increased ( t =2.621,P =0.012; t =-3.714,P =0.001 ; t =17.595,P=0.000).In aM CI group,numbers of long range fiber in right frontal lobe were reduced (2895 ± 343 vs.3451 -± 230,t =- 7.011,P =0.000),and ADC values were increased ( t =4.443,P =0.000).In aMCI group,numbers of long range fiber in left frontal lobe were positively correlated with scores of mini-mental state examination ( MMSE ) ( r =0.457,P =0.015 ),while ADC values of long range fiber in left frontal lobe were negatively correlated with scores of MMSE ( r=-0.415,P=0.028).ConclusionThe structure and fiber connectivity are affected in aMCI patients and the lesion of connectivity in left frontal lobe are related to the severity of symptom.

Objective: To investigate the current status of screen time in preschoolers during the outbreak of Corona Virus Disease 2019 (COVID-19), and the influences of parent-child active play and school online education on screen time. Methods: A total of 2 370 preschoolers were chosen through clustering sampling from 7 kindergartens in Wuxi urban areas. Questionnaire survey was administered to parents for child screen time, active play, online education as well as characteristics information. Results: A total of 1 428(60.3%) preschoolers reported more than 1 hour screen time a day, and 1 915(80.8%) preschoolers reported more than 20 minutes consecutive screen time. Compared with the weekend before the COVID-19 outbreak, 1 551(65.4%) preschoolers reported an increase of 10 minutes in screen time perday, and 1 444(60.9%) reported an increase of 5 minutes for one sersion. Logistic regression analysis showed that after controlling for confounding factors, high frequency of parent-child active play, parents’ exercise habits, online education from kindergartens and children’s active participation in the online education were negatively associated with preschoolers’ excessive and increased screen time during this period(OR=0.39-0.79, P<0.05). Conclusion During the COVID-19 outbreak, preschoolers’ media use in Wuxi city is worrisome. Parent-child active play, parents’ exercise and online education from kindergartens may play a positive role in reducing preschoolers’ media use.

Objective:To explore the model of first aid skills training based on competition.Method:The second-year residents who participated in the competition in 2021 ( n = 142) were selected. According to whether they attended BLS training in the first year, they were divided into group A ( n = 88) who attended BLS training and group B ( n = 54) who did not. Chi-square test, T test, Wilcoxon rank sum test and Logistic regression were used in our study. Results:There was no significant difference in gender and specialty between both groups (female, 62.5% vs. 68.5%, P= 0.466; TCM, 50% vs. 53.7%, P= 0.668), but the real resuscitation experience of group A was more than that of group B (40.9% vs. 9.3%, P= 0.000). The qualified rate of CPR and the compression score in group A was higher than that in group B [(81.8% vs. 61.1%, P = 0.006; (30±5) vs. (25 ±10), P= 0.001], including compression frequency, depth, rebound and compression/respiration rate (73.9% vs. 55.6%, P= 0.024; 88.6% vs. 70.4%, P= 0.006; 96.6% vs. 87%, P= 0.031). In the theoretical examination, the correct rates of electrocardiogram [(53.63±2.9)% vs. (50.44±2.57)%] and first aid medication [(57.38±3.55)%, P = 0.001] in the two groups were significantly lower than the qualified rate. After adjusting other factors, Logistic regression analysis showed that the CPR qualification rate in group A was 2.769 times higher than that in group B ( P= 0.015, 95% CI 1.215~6.311) Conclusions:The first aid skills training mode based on competition can objectively reflect first aid skills level of residents. We found that the quality of CPR skills was not related to gender, specialty and real CPR experiences but experience of BLS training was an independent influencing factor. And how to identify and manage arrhythmias and how to choose different emergency drugs are urgent emergency skills for residents to improve.

Objective To investigate the effect of preoperative oral midazolam on postoperative de-lirium in elderly patients with preoperative moderate-severe anxiety undergoing radical resection of colorectal cancer.Methods Eighty elderly patients undergoing laparoscopic surgery for radical resection of colorectal cancer,32 males and 48 females,aged 65-79 years,BMI 21-27 kg/m2,ASA physical status Ⅱ or Ⅲ,the state-trait anxiety inventory(STAI-S)≥38 scores at admission were selected.Patients were divided into two groups using random number method:control group and midazolam group,40 patients in each group.The midazolam group were administrated midazolam 7.5 mg per night till one day before surgery,while the placebo was administrated in the control group.The incidence of delirium 3 days after surgery and the STAI-S scores of one day before surgery were evaluated.The HR and MAP at entry,30 minutes after an-esthesia induction,1 hour,2 hours after anesthesia induction,and 30 minutes after extubation were recor-ded.The total dose of propofol,remifentanil and dexmedetomidine and the using rate of metaraminol were recorded.The visual analog scale scores 30 minutes after extubation,24 and 72 hours after surgery,the u-sing rate of tramadol,and the extubation time were recorded.Results Compared with the control group,the STAI-S scores of one day before surgery,and the incidence of postoperative delirium,the rate of using metaraminol,the VAS scores 30 minutes after extubation and 24 hours after surgery,the rate using of tram-adol were significantly decreased in the midazolam group(P＜0.05).There were no significant differences in total dose of propofol,remifentanil,and dexmedetomidine,extubation time between the two groups.Conclusion Preoperative oral midazolam can effectively reduce the incidence of postoperative delirium in elderly patients with preoperative moderate-severe anxiety undergoing radical resection of colorectal cancer.

Objective:To elucidate the drug resistance characteristics and molecular epidemiological features of carbapenem-resistant Klebsiella pneumoniae (CRKP) in this region, and to provide a basis for clinical anti-infective therapy. Methods:A total of 92 CRKP isolates were collected from the First Affiliated Hospital of Ningbo University and Ningbo No.2 Hospital. Antimicrobial susceptibility testing was performed using the VITEK2 Compact analyzer. Whole-genome sequencing was used to analyze their molecular characteristics including capsular serotypes, sequence types (multilocus sequencing typing) and resistance genes. Plasmid replicon types were identified using PlasmidFinder, and core genome phylogenetic analysis was performed with Parsnp.Results:The CRKP isolates were mainly isolated from critically ill elderly patients, with sputum (51.09%, 47/92) and urine (13.04%, 12/92) as the major specimen sources, followed by blood (8.70%, 8/92). Most of the infections were reported in the intensive care unit (33.70%, 31/92). All isolates showed high resistance to commonly used antibiotics, but lower resistance to polymyxin B (6.52%, 6/92). The predominant resistance gene, capsule serotype and ST were blaKPC-2 (78.26%, 72/92), KL64 (48.91%, 45/92), and ST11 (54.35%, 50/92), respectively. ST11-KL64 was the predominant clone, accounting for 46.74%(43/92) and carried multiple plasmids with IncR and IncFⅡ as the major types. Conclusions:CRKP in this region mainly harbored the blaKPC-2 resistance gene and showed high resistance to commonly used antibiotics. The ST11-KL64 clone spread widely in this region and the strains circulating in the two hospitals were similar. Thus, the surveillance should be strengthened.

Objective:This work aims to assess the distribution of peripheral blood monocyte subsets, the expression level of the functional markers in rheumatoid arthritis (RA) patients, and analyze the correlation between the above indexes and the onset of RA.Methods:Peripheral blood mononuclear cells were collected and isolated from 62 RA patients, 52 healthy control (HC) and 12 disease control group′s patients via density centrifugation. The enrolled patients were attended or underwent physical examination in East Hospital, Tongji University from June 2020 to December 2021. Monocytes could be classified into classical (CM), intermediate (IM) and non-classical (NCM). Then, the flow cytometry was performed to examine the distribution of monocyte subsets and the measure the expression level of human leukocyte antigen DR (HLA-DR), intracellular tumor necrosis factor α (TNF-α) in peripheral blood monocytes. The statistical methods in this study mainly include: Kruskal-Wallis H test, Chi-Square test, Mann-Whitney U test, Wilcoxon matched-pairs signed ranks test, Spearman correlation coefficient test and Logistic regression analysis. The diagnostic value of IM proportion in RA was analyzed by ROC curve. Results:The monocytes number and monocytes proportion in white blood cells were much higher in RA [0.40 (0.40, 0.50), 7.60% (5.97%, 8.53%)] and disease control [0.40 (0.40, 0.68), 8.20% (5.85%, 10.28%)] compared with HC [0.30 (0.30, 0.40), 5.80% (5.03%, 6.38%)] ( H=24.733, P<0.001; H=27.469, P<0.001). A statistic-significant difference was detected among the proportion of CM[85.49%(76.91%,89.21%),88.94%(86.36%,91.72%),90.26%(80.25%, 92.56%)],IM[11.65%(8.47%,17.89%),7.89%(5.36%,10.75%), 5.56%(4.17%, 8.27%)], NCM[2.22%(1.39%, 3.74%), 2.49%(1.74%, 4.66%), 5.13%(3.39%, 9.85%)] in RA group, HC group and disease control group ( H=11.389, P=0.003; H=20.815, P<0.001; H=10.640, P=0.005). The proportion of CM was lower in RA and the IM proportion was increased in RA( P=0.003; P=0.003). The intracellular TNF-α level of monocytes in all three groups revealed the trend that IM>NCM>CM. The intracellular TNF-α in IM of RA was positively associated with serum TNF-α ( r=0.376, P=0.041). The HLA-DR expression in IM subsets were higher than CM and NCM subsets in all RA,HC and disease control groups. The expression of HLA-DR of IM in RA group and disease control was higher than HC group [8 611.50 (6201.3, 9890.8), 10 295.0 (7 899.0, 13632.0), 6 278.00(4 057.8, 9522.0), H=10.495, P=0.005]. There were no correlations between the proportion of peripheral blood IM and clinical characteristics CRP ( r=0.119, P=0.359), RF ( r=0.204, P=0.112) and ESR ( r=0.153, P=0.236). Logistic regression analysis showed that the proportion of IM ( OR=1.169, 95% CI 1.003-1.363, P=0.046), CRP ( OR=1.277, 95% CI 1.000-1.631, P=0.050), RF ( OR=1.179, 95% CI 1.080-1.287, P<0.001) are positively correlated with RA onset. The area under ROC curve for diagnosis of RA with IM proportion was 0.687, and the 95% confidence interval was 0.590-0.784, P<0.001. Conclusions:The distribution of monocyte subsets in peripheral blood of RA patients is abnormal. The increase in the proportion of IM, the enhanced antigen-presenting ability, and the increased level of TNF-α secretion in RA patients may play an important role in the pathogenesis of RA.

Objective To analyze the mortality rate in index partners of HIV discordant couples in the context of antiretroviral therapy and related factors in Zhumadian,Henan province.Methods An open and prospective cohort survey was conducted annually during 2008-2014.Information about the index partners'demographic characteristics,CD4+T (CD4) cell counts,viral load,AIDS-related syndromes,date of therapy initiation,level of treatment institution,and outcome of death were collected.Life table was used to calculate survival rate,and Cox proportional hazards model was used to analyze the mortality related factors.Results Among 4 196 index partners receiving antiretroviral therapy between 2008 and 2014,350 were fatal,with the mortality rate of 1.88 per 100 person years (95％CI:1.86-1.90).The survival rates of these patients with the duration of follow-up from 1 to 7 years were 99.64％,97.70％,95.62％,93.54％,91.25％,86.86％ and 82.36％ respectively.Factors related to the survival of the index partners of HIV discordant couples were age ≥50 years (HR=3.48,95％CI:2.67-4.55),junior high school or above education level (HR=0.75,95％CI:0.56-1.00),baseline CD4 cell counts ＞350 cells per μl (HR=0.55,95％CI:0.42-0.72),baseline viral load ＞400 copies/ml (HR=1.71,95％CI:1.32-2.22) and initiating treatment prior to cohort entry (HR=0.65,95％ CI:0.41-1.05).Conclusions The mortality rate of index partners receiving antiretroviral therapy was relatively low,indicating a better effect of long-term treatment on patients' survival.It is necessary to further identify and control the key factors influencing the survival of patients receiving therapy,improve the timeliness and adherence of the treatment,keeping patients' immunity level.Meanwhile,much attention should be paid to the drug resistance due to long term treatment and treatment failure.