Objective:To explore the clinical characteristics and molecular basis for children and adolescents with monogenic diabetes.Methods:A retrospective analysis was carried out for the clinical manifestations and laboratory data of 116 children and adolescents diagnosed with diabetes at Ningbo Women and Children′s Hospital from January 2020 to March 2023. Whole exome sequencing and mitochondrial gene sequencing were carried out on 21 children with suspected monogenic diabetes.Results:A total of 10 cases of monogenic diabetes were diagnosed, all of which were Maturity-onset Diabetes Of the Young (MODY). Six cases of MODY2 were due to GCK gene mutations, 1 case of MODY3 was due to HNF1A gene mutation, 2 cases of MODY12 were due to ABCC8 gene mutations, and 1 case of MODY13 was due to KCNJ11 gene mutation. Nine of the 10 patients with MODY had no typical symptoms of diabetes. A family history of diabetes was significantly more common in the MODY group compared with the T1DM and T2DM groups ( P<0.05). The BMI of the MODY group was higher than that of the T1DM group ( P<0.05). The initial blood glucose level was lower than that of the T1DM group ( P<0.05), and there was no significant difference compared with the T2DM group. The fasting C-peptide level of the MODY group was higher than that of the T1DM group ( P<0.05), and there was no significant difference compared with the T2DM group. Glycosylated hemoglobin of the MODY group was lower than both the T1DM and T2DM groups ( P<0.05). Conclusion:In this study, MODY has accounted for the majority of monogenic diabetes among children and adolescents, and the common mutations were those of the GCK gene in association with MODY2. Blood glucose and glycosylated hemoglobin of children with MODY were slightly increased, whilst the islet cell function had remained, and the clinical manifestations and laboratory tests had overlapped with those of type 2 diabetes. WES and mitochondrial gene sequencing can clarify the etiology of monogenic diabetes and facilitate precise treatment.

Objective:To construct the evaluation index system of core competence of specialized gynecological nurses, and to provide reference for the curriculum setting and effect evaluation of standardized training for specialized gynecological nurses.Methods:The draft evaluation index system was prepared through literature research and group discussion. From May to June 2021, two rounds of Delphi letter consultation was used among 18 experts, to complete the selection and modification of evaluation index, and to determine the index weight.Results:The effective response rate of the two rounds of Delphi expert consultation was 100%, the expert authority coefficient was 0.938, and the Kendall harmony coefficient of the two rounds of expert letter consultation was 0.117 and 0.304, respectively (both P<0.01). Finally, the evaluation index system of core competence of specialized gynecological nurses was formed, including 4 first-level indexes, 11 second-level indexes and 48 third-level indexes. Conclusions:The evaluation index system of core competence of specialized gynecological nurses established in this study is scientific and reliable, which can provide reference for the curriculum setting and effect evaluation of standardized training for specialized gynecological nurses.

Objective:To explore the effects of magnanimous therapy on the magnanimous and enterprising traits of lung cancer patients and the analysis of related factors.Methods:Totally 197 patients with lung cancer were divided into individual group ( n=62), team group ( n=75) and control group ( n=60). Comparison and correlation analysis were applied to the data before and after the electroencephalogram and the magnanimous questionnaire, the cancer response questionnaire, the T-type psychological scale, the cancer heart state questionnaire and the cancer patient's life function index scale. t test, analysis of variance and Pearson correlation analysis were processed by SPSS 23.0. Results:After treatment, the " enterprising" dimension and " magnanimous" dimension of individual group and the " enterprising" dimension of the team group ((3.035±0.309), (3.041±0.265), (3.173±0.371)) were higher than that before treatment((2.934±0.326), (2.908±0.315), (3.130±0.387), all P<0.05). There was negative correlation between " magnanimous" dimension of the magnanimous questionnaire and " subconscious" dimension of the T-type psychological scale in individual group( r=-0.280, P<0.05). In team group, the " enterprising" dimension of the magnanimous questionnaire was negatively correlated with " Psychological" and " Yield" dimension of the cancer heart state questionnaire( r=-0.279, -0.285, P<0.05), and positively correlated with " Facing" of the cancer response questionnaire, " Good physical condition and ability" and " Psychological well-being" dimension of the cancer patient's life function index scale( r=0.367, 0.402, 0.379, P<0.05). There was a negative correlation between the " enterprising" dimension of the magnanimous questionnaire and the beta wave value in individual group. Conclusion:The magnanimous therapy can improve enterprising and magnanimous level of patients with lung cancer, and the effects are related with the above-mentioned psychosomatic factors.

Objective To investigate the effect of targeted silencing Notch1 on proliferation and apoptosis of human non-small cell lung cancer stem cells.Methods Lung cancer A549 cells and SPC-A-1 cells were selected and divided into control group,Nc-shRNA group and Notch1-shRNA group.The Nc-shRNA group was a negative control RNAi lentivirus group,and the Notch1-shRNA group was a Notch1 inhibitory RNAi lentivirus group.The lentiviral-mediated shRNA interference technology was used to target the silencing of Notch1.The silencing effect of Notch1 gene was verified by quantitative real time polymerase chain reaction (qRT-PCR) and Western blotting.Cell proliferation was detected by methyl thiazolyl tetrazolium (MTT) and sarcosphere formation assay.Apoptosis was detected by Annexin V/7-AAD double staining.Western blotting was used to detect the expression of proliferating cell nuclear antigen (PCNA),B-cell lymphoma-2 (Bcl-2) and Notch1 downstream gene Hes-1.Results The results of qRT-PCR showed that the relative expression levels of Notch1 in control group,Nc-shRNA group and Notch1-shRNA group in A549 cells and SPC-A-1 cells were 1.000 ± 0.000,0.937 ± 0.025,0.490 ± 0.036 and 1.000 ± 0.000,1.077 ± 0.070,0.373± 0.038,with statistically significant differences (F =359.707,P ＜0.001;F =210.455,P ＜0.001),further paired comparison,the relative expression of Notch1 in Notch1-shRNA group was significantly lower than that in Nc-shRNA group (all P ＜ 0.05).Western blotting showed that the expressions of Notch1 protein in A549 cells and SPC-A-1 cells were consistent with the mRNA results.MTT assay showed that the 24 h A values of A549 cells in control group,Nc-shRNA group and Notch1-shRNA group were 0.209 ± 0.005,0.219 ± 0.009,0.159 ±0.006,48 h A values were 0.293 ± 0.004,0.302 ± 0.004,0.205 ± 0.005,72 h A values were 0.450 ± 0.003,0.430 ± 0.012,0.348 ± 0.017,with statistically significant differences (F =79.487,P＜0.001;F =508.664,P ＜0.001;F =57.156,P ＜0.001),further paired comparison,the proliferation ability of Notch1-shRNA group was significantly lower than that of Nc-shRNA group at 24,48,72 h (all P ＜ 0.05).The 48 h A values of SPC-A-1 cells in control group,Nc-shRNA group and Notch1-shRNA group were 0.438 ±0.022,0.412 ± 0.015,0.364 ± 0.010,72h A values were 0.540 ± 0.016,0.519 ± 0.009,0.438 ± 0.019,with statistically significant differences (F =15.667,P =0.004;F =37.299,P ＜ 0.001),further paired comparison,the proliferation ability of Notch1-shRNA group was significantly lower than that of Nc-shRNA group at 48 h and 72 h (all P ＜ 0.05).The sphere sizes of control group,Nc-shRNA group and Notch1-shRNA group in A549 cells were (149.667 ± 6.506) μm,(136.667 ± 7.095) μm,(86.676 ± 7.638) μm,with statistically significant difference (F =65.940,P ＜ 0.001).The sphere sizes of the three groups in SPC-A-1 cells were (118.667 ± 6.658) μm,(128.000 ± 7.000) μm,(60.675 ± 4.509) μm,with statistically significant difference (F =105.372,P ＜0.001).Further paired comparison,the sphere size of Notch1shRNA group was significandy smaller than that of Nc-shRNA group in the two kinds of cells (all P ＜ 0.05).The apoptosis rates of control group,Nc-shRNA group and Notch1-shRNA group in A549 cells and SPC-A-1cells were (0.489 ± 0.014)％,(0.633 ± 0.021)％,(1.683 ± 0.221)％ and (1.323 ± 0.194)％,(1.690 ± 0.188) ％,(3.017 ± 0.356) ％,with statistically significant differences (F =77.660,P ＜ 0.001;F=32.200,P =0.001),further paired comparison,the apoptosis rate of Notch1-shRNA group was significantly higher than that of Nc-shRNA group in the two kinds of cells (all P ＜ 0.05).Western blotting showed that the expressions of PCNA,Bcl-2 and Hes-1 in control group,Nc-shRNA group and Notch1-shRNA group in A549 cells were statistically significant (F =155.343,P ＜ 0.001;F =22.576,P =0.002;F =70.108,P＜0.001),and the expressions of PCNA,Bcl-2 and Hes-1 in the three groups in SPC-A-1 cells were statistically significant (F =49.419,P ＜0.001;F =28.090,P =0.001;F =12.040,P =0.007).Further paired comparison,the expressions of PCNA,Bcl-2 and Hes-1 in Notch1-shRNA group were significantly lower than those in Nc-shRNA group in the two kinds of cells,and the differences were statistically significant (all P ＜0.05).Conclusion Targeted silencing of Notch1 can reduce the proliferation activity of lung cancer stem cells and promote apoptosis,which may be related to the down-regulation of its downstream gene Hes-1.

OBJECTIVE:To establish a method for the content determination of loxoprofen silver in Loxoprofen silver cream, and provide reference for the quality control of the preparation. METHODS:HPLC was performed on the column of Diamonsil C18 with mobile phase of methanol-water-glacial acetic acid-triethylamine(55:45:0.1:0.1,V/V/V/V)at a flow rate of 1.0 mL/min,de-tection wavelength was 223 nm,column temperature was 30 ℃,and the injection volume was 20 μL. RESULTS:The linear range of loxoprofen silver was 6.53-130.7μg/mL(r=0.9999);the limit of quantitation was 0.253μg/mL,limit of detection was 0.076μg/mL;RSDs of precision,stability and reproducibility tests were lower than 2%;recovery was 96.79%-103.68%(RSD=2.23%, n=9). CONCLUSIONS:The method is simple,accurate and reproducible,and can be used for the content determination of Loxo-profen silver cream.

Objective To study the physicochemical properties of aqueous solution of surface active drug montelukast so-dium (MS) ,which could provide experimental basis for further development of micelle or mixed micelle preparations .Methods Critical micelle concentration (CMC) of MS at different temperatures were determined by conductivity measurements .The absorbance and transmittance of MS aqueous solution were measured by UV at different sodium chloride concentration levels . The micelle stability was evaluated via high speed centrifugal .Results The CMC of MS aqueous solution at 25℃ ,30℃ ,35℃were 0 .75 ,0 .82 ,0 .90 mmol/L .The absorbance and transmittance of MS aqueous solution were affected by the sodium chlo-ride concentration and the concentration of MS itself .It was observed that a clear solution was obtained when MS concentration>7 .5 mmol/L and no precipitation was noticed even after high speed centrifugal .Conclusion Montelukast sodium is a surface active drug .Its solubility is related to MS concentration .The solubility is also sensitive to the temperature and the electrolyte concentration .These unique physicochemical properties could be used to develop micelle or mixed micelle pharmaceutical prepa-rations .

Objective: To evaluate the clinical characteristics, MYD88^L265P mutation, CXCR4^WHIM mutation and prognosis in patients with Waldenström macroglobulinemia (WM). Methods: The clinical characteristics, International Prognostic Scoring System for symptomatic WM (WPSS) , and overall survival (OS) were retrospectively assayed in 93 patients with newly diagnosed WM at Peking Union Medical College Hospital during January 2000 to August 2016. The MYD88^L265P mutation and CXCR4^WHIM mutation were tested among 34 patients. Results: The median age of the 93 patients was 64 years (range, 33-85 years) with a male-to-female ratio of 2.44. According to WPSS, we included 16 (17.2%) low-risk, 44 (47.3%) intermediate-risk and 33 (35.5%) high-risk patients. Eight patients had secondary amyloidosis. With a median follow-up of 44 (1-201) months, the median OS was 84 months. Cox regression multifactor analysis showed WPSS risk group (HR=2.342, 95% CI 1.111-4.950, P=0.025) , whether patients had secondary amyloidosis (HR=5.538, 95% CI 1.958-15.662, P=0.001) and whether patients received new drugs (HR=3.392, 95% CI 1.531-7.513, P=0.003) were independent factors associated with OS. We have investigated the presence of the MYD88^L265P and CXCR4^WHIM mutation in 34 patients and found that MYD88^L265P mutation was occurred in 32 patients (94.1%) and CXCR4^WHIM mutation was occurred in 8 patients (23.5%). Seven of 8 patients who harbored CXCR4^WHIM-mutated also exhibited the MYD88^L265P mutation. Patients with MYD88^L265PCXCR4^WHIM vs MYD88^L265PCXCR4^WT presented with more severe anemia, lower platelet level, higher M protein level and more hyper-viscosity syndrome. Conclusion WPSS risk group, whether patients had secondary amyloidosis or received new drugs are independent factors for OS in WM. MYD88^L265P and CXCR4^WHIM mutation, the most common somatic variants in WM, often occur together and impact the clinical presentation.

Objective To observe the biological characteristics of db/db mice, and to provide the basis for application of db/db mice in experimental research.Methods Spontaneous type 2 diabetes BKS.Cg-Dock7m +/+ Leprdb/JNju mice and wild type mice of the same age were used in this study.Their fasting blood glucose was determined at 8,12,16, 20 and 24 weeks of age, the body weight was recorded at 10, 12,16, 20 and 24 weeks of age, the levels of serum insulin, total cholesterol and triglyceride were measured, the organ weight and liver coefficient were determined, and the liver and pancreas were taken for pathological examination at 24 weeks of age.Results The db/db mice maintained a high level of fasting blood glucose and body weight.Levels of serum insulin, total cholesterol, and triglyceride were significantly higher than the wild type mice.Compared with wild type mice, liver weight and kidney weight were also significantly increased.Obvious pathological changes of liver and pancreas were observed in 24-week old db/db mice.Conclusions db/db mouse has obvious characteristics of type 2 diabetes, such as hyperglycemia, hyperlipidemia, and hyperinsulinemia, can maintain stable levels of high blood glucose,and is an ideal animal model for experimental study of type 2 diabetes mellitus.

Aim To investigate the proliferative effect and the apoptosis of rat epididymal epithelial cells induced by podophyllotoxin and its underlying mechanisms.Methods Primary epididymal epithelial cells were cultured in vitro.CCK-8 assay was used to analyze proliferation of epididymal epithelial cells induced by podophyllotoxin on 24, 48 and 72 h.The ultra structural changes of the epididymal epithelial cells were observed by transmission electron microscope.AnnexinV-FITC/PI staining was used to quantify the percentages of apoptosis in the total cell population.The TdTmediated dUTP nick end labeling(TUNEL) technique was applied to observe the morphological changes of apoptotic cells.The expression of tumor necrosis factor alpha(TNF-α) mRNA was investigated by real-time RT-PCR.The level of TNF-α in cell culture supernatant was measured by enzyme-linked immunosorbent assay(ELISA) technology.Western blot was per-formed to determine the protein expression of cytochrome C, caspase-3, caspase-8, caspase-9.Results Podophyllotoxin significantly inhibited the activity of proliferation and induced apoptosis of epididymal epithelial cells in a dose-and time-dependent manner(P<0.05), with a 50％ inhibitory concentration(IC50) value and corresponding 95％ confidence intervals(CI) of 59.36(15.50～227.41), 0.37(0.080～1.70), 0.077(0.017～0.35) μmol·L-1 at 24, 48 and 72 h, respectively.Podophyllotoxin induced cell volume turned round and cell nuclear fragmentation and mitochondrial vacuolation.RT-PCR and ELISA results showed that podophyllotoxin improved the expression of TNF-α mRNA and protein.Western blot results demonstrated that podophyllotoxin activated the protein expression of cytochrome C, caspase-3, caspase-8 and caspase-9.Conclusion Podophyllotoxin can induce rat epididymal epithelial cell apoptosis through both the mitochondria-regulated intrinsic pathway and the TNF receptor-mediated extrinsic pathway.

Objective To discuss the influence of family function and social support on insulin treatment compliance in patients with diabetes mellitus. Methods A total of 368 diabetic patients diagnosed using insulin treatment were selected between March 2013 and May 2014 in outpatient department of endocrinology department. Situations of Insulin treatment compliance, family function, social support were investigated by questionnaire. Results Insulin treatment compliance were good in 202 cases (54. 9%), poor in 120 cases (32. 6%) and very poor in 46 cases (12. 5%). The cases of different levels of family function were 183 (49. 7%) with a high level, 82 (22. 3%) with a general level and 103 (28. 0%) with dereliction of duty level. The cases of low, a general and a high level of social support were 119 (32. 3%), 108 (29. 3%) and 141 (38. 3%). Multiple regression analysis showed that the main factors influencing the compliance of insulin treatment were age, education degree, marital status, occupation, monthly income, payment methods, disease duration, family function and social support. Conclusions When the scores of family function and social support are higher, the treatment compliance is higher too. The nursing staff should improve patient's family function and social support in order to improve the patients' insulin treatment compliance, so as to improve patients'quality of life.