ObjectiveTo investigate the status of tooth loss in people aged 50 and above, so as to understand their oral health status and provide scientific evidences for promoting oral health of middle-aged and elderly people. MethodsA total of 400 patients who visited the department of stomatology at Sijing Hospital in Songjiang District of Shanghai were performed oral health examinations and their information was collected according to the national epidemiological survey standards for oral health. ResultsThere were statistically significant differences in tooth loss among people aged 50 and above with different ages, educational levels, occupations, types of medical insurance and chronic diseases (P<0.05), but gender and monthly income had no statistically significant correlations with tooth loss (P>0.05). Among lifestyle factors, smoking, alcohol consumption and tea drinking had no statistically significant impacts on the number of remaining teeth (P>0.05), but toothbrushing frequency, flossing frequency, toothpick use frequency, toothbrush replacement frequency, and tooth loosening were statistically associated with the number of remaining teeth (P<0.05). Multiple linear regression analyses indicated that a total of 7 related factors including age, educational level, occupation, medical payment type, chronic disease, tooth loosening and toothpick use frequency were significantly associated with the number level of remaining teeth in individuals aged 50 and above. ConclusionAge, chronic disease, and tooth loosening were influencing factors affecting the number of teeth left in people aged 50 and above. It is recommended to strengthen oral health education and improve healthcare awareness to reduce the risk of tooth loss in people aged 50 and above.

Objective The present study was designed to explore the inhibitory effects of the ADC drug Disitamab Vedotin(RC-48)on breast cancer cells with different HER-2 expression levels by utilizing a tumor organoid culture system.Methods Within the framework of the tumor organoid culture system,the breast cancer cell lines MCF-7(characterized by low HER-2 expression,Luminal A subtype)and BT-474(exhibiting high HER-2 expression,HER-2 positive subtype)were cultured independently and in various mixed ratios.The histological characteristics,as well as the expression levels and distribution of HER-2 in MCF-7 and BT-474 organoids,were analyzed via immunohistochemistry and immunofluorescence techniques.MCF-7 and BT-474 organoids were separately treated with Vedotin(RC-48),Disitamab,and Monomethyl auristatin E(MMAE).Additionally,a drug sensitivity test of Disitamab Vedotin(RC-48)was carried out on mixed MCF-7 and BT-474 cell ratios and on patient-derived breast cancer organoids,with the assessment conducted using the 3D-Glo method.Results In the tumor organoid culture system,immunohistochemistry and immunofluorescence analyses demonstrated that HER-2 was predominantly localized in the cell membrane.Specifically,BT-474 organoids exhibited robust HER-2 expression,while MCF-7 organoids displayed relatively low expression levels.When compared with MCF-7 organoids,RC48-ADC exerted a more pronounced inhibitory effect on BT-474 organoids,with IC50 values of 109.7 μg/mL and 2.792 μg/mL,respectively.The co-culture model further confirmed the bystander effect of RC-48,revealing that the ratio of HER-2-positive to HER-2-negative cells significantly influenced drug efficacy.Additionally,treatment with RC-48 led to a reduction in HER-2 expression in breast cancer organoids with diverse HER-2 expression levels.Conclusions The tumor organoid model can accurately mirror drug sensitivity and bystander effects.Within this model,RC-48 effectively inhibited HER-2 highly-expressing breast cancer cells,augmented the killing effect through the bystander mechanism,and downregulated HER-2 expression,thereby suggesting its potential for targeting HER2-associated breast cancer.

Objective To investigate the status of oral frailty(OF)in patients who underwent chemotherapy for lung cancer,identify key factors influencing OF,and develop a risk prediction model.Methods Using convenience sampling,431 lung cancer inpatient were recruited from three Tier-IIIA hospitals in Jiangsu Province between September and November 2024 as the training cohort.The patients were divided into OF and non-OF groups.Relevant data were compared between the two groups.Multifactorial logistic regression analysis was performed to determine factors that associated with OF,and a risk prediction model was created accordingly.Receiver operating characteristic(ROC)curve analysis was used to predict model performance.In December 2024,additional 185 patients from one other Tier-IIIA hospitals were recruited to validate the developed model.Results The prevalence of OF among lung-cancer patients undergoing chemotherapy was 58.93%.Following listed items were identified as the risk factors of OF(all P<0.05):older in age(OR=3.420),poor education(OR=0.030),brain metastasis(OR=7.880),high nutritional risk screening 2002 score(OR=1.550),elevated C-reactive protein(OR=1.100),and elevated lactate dehydrogenase(OR=1.010).ROC area under the curve(AUC)of the model was 0.860(95%CI:0.830-0.900)in modelling cohort and 0.840(95%CI:0.780-0.900)in validation cohort.Hosmer-Lemeshow goodness-of-fit test yielded χ 2=4.870,P=0.770 for the training set and χ 2=2.770,P=0.950 for the validation set.Conclusion The risk prediction model for OF developed in this study demonstrates a good predictive performance and can facilitate early identification of high-risk patients,thereby providing a scientific basis for clinical interventions.

Objective To investigate the status of oral frailty(OF)in patients who underwent chemotherapy for lung cancer,identify key factors influencing OF,and develop a risk prediction model.Methods Using convenience sampling,431 lung cancer inpatient were recruited from three Tier-IIIA hospitals in Jiangsu Province between September and November 2024 as the training cohort.The patients were divided into OF and non-OF groups.Relevant data were compared between the two groups.Multifactorial logistic regression analysis was performed to determine factors that associated with OF,and a risk prediction model was created accordingly.Receiver operating characteristic(ROC)curve analysis was used to predict model performance.In December 2024,additional 185 patients from one other Tier-IIIA hospitals were recruited to validate the developed model.Results The prevalence of OF among lung-cancer patients undergoing chemotherapy was 58.93%.Following listed items were identified as the risk factors of OF(all P<0.05):older in age(OR=3.420),poor education(OR=0.030),brain metastasis(OR=7.880),high nutritional risk screening 2002 score(OR=1.550),elevated C-reactive protein(OR=1.100),and elevated lactate dehydrogenase(OR=1.010).ROC area under the curve(AUC)of the model was 0.860(95%CI:0.830-0.900)in modelling cohort and 0.840(95%CI:0.780-0.900)in validation cohort.Hosmer-Lemeshow goodness-of-fit test yielded χ 2=4.870,P=0.770 for the training set and χ 2=2.770,P=0.950 for the validation set.Conclusion The risk prediction model for OF developed in this study demonstrates a good predictive performance and can facilitate early identification of high-risk patients,thereby providing a scientific basis for clinical interventions.

Objective The present study was designed to explore the inhibitory effects of the ADC drug Disitamab Vedotin(RC-48)on breast cancer cells with different HER-2 expression levels by utilizing a tumor organoid culture system.Methods Within the framework of the tumor organoid culture system,the breast cancer cell lines MCF-7(characterized by low HER-2 expression,Luminal A subtype)and BT-474(exhibiting high HER-2 expression,HER-2 positive subtype)were cultured independently and in various mixed ratios.The histological characteristics,as well as the expression levels and distribution of HER-2 in MCF-7 and BT-474 organoids,were analyzed via immunohistochemistry and immunofluorescence techniques.MCF-7 and BT-474 organoids were separately treated with Vedotin(RC-48),Disitamab,and Monomethyl auristatin E(MMAE).Additionally,a drug sensitivity test of Disitamab Vedotin(RC-48)was carried out on mixed MCF-7 and BT-474 cell ratios and on patient-derived breast cancer organoids,with the assessment conducted using the 3D-Glo method.Results In the tumor organoid culture system,immunohistochemistry and immunofluorescence analyses demonstrated that HER-2 was predominantly localized in the cell membrane.Specifically,BT-474 organoids exhibited robust HER-2 expression,while MCF-7 organoids displayed relatively low expression levels.When compared with MCF-7 organoids,RC48-ADC exerted a more pronounced inhibitory effect on BT-474 organoids,with IC50 values of 109.7 μg/mL and 2.792 μg/mL,respectively.The co-culture model further confirmed the bystander effect of RC-48,revealing that the ratio of HER-2-positive to HER-2-negative cells significantly influenced drug efficacy.Additionally,treatment with RC-48 led to a reduction in HER-2 expression in breast cancer organoids with diverse HER-2 expression levels.Conclusions The tumor organoid model can accurately mirror drug sensitivity and bystander effects.Within this model,RC-48 effectively inhibited HER-2 highly-expressing breast cancer cells,augmented the killing effect through the bystander mechanism,and downregulated HER-2 expression,thereby suggesting its potential for targeting HER2-associated breast cancer.

Objective In cases and events of mixed STR typing of aborted fetus,two methods for calculating paternity index(PI)of suspected biological fathers are proposed,which could be useful for theoretical reference for parental identification including mixed STR typing.Methods Depending on whether the fetal genotypes can be identified,the simple PI calculation method and the PI calculation method of deduced biological paternal genes when the fetal genotypes cannot be identified are proposed.Results The simple PI calculation method is to indentify the fetal genotypes first and then calculate according to the standard triplet.The PI calculation method of deduced biological paternal genes is to deduce all the possible genotypes of biological fathers conforming to Mendel's law(inference)without considering the ratio of peak height and peak area in mixed typing,and then calculate the parental index separately,taking the minimum value as the parental index of the locus.Conclusion When mixture ratio of fetus in the mixed typing of aborted tissue MR≥0.43,the accuracy of separation is very high and the simple PI calculation method can be accurate,so it is recommended.If 0.05≤MR＜0.43,it is suggested to use the calculation method of deduced biological paternal genes,which can avoid misjudgment of irrelevant persons to the greatest extent.If MR＜0.05,there's a high risk of fetal allele loss,we should not perform a paternity test on the mixed spot.Since the cumulative parental index calculated by deduced the biological paternal genes is usually lower than the value calculated by dividing the fetal genotype,the CPI may be lower than 10 000 when fewer loci are identified,and then more genetic markers should be detected.

Objective To investigate the risk factors of severe complications of elderly patients with pancreaticoduodenectomy (PD). Methods The clinical data of 399 elderly patients with PD and 65 years or more (65-91years) including 76 cases with 75 years or more （19.0%） in this hospital from January 2016 to December 2020, were retrospectively analyzed. Less Barthel index score was used to indicate lower activities of daily living. The risk factors of severe complications after PD were analyzed by binary Logistic regression and predictive model was established. Receiver operating characteristic (ROC) curve was used to verify the predictive efficiency of model. Results A total of 224 cases were found postoperative complications from 399 elderly PD cases with the rate 56.14% including 187 (46.87%) cases of mild complications and 37 (9.27%) cases of severe complications. Age ≥75 years (OR=2.78, P<0.05), combined with hypertension (OR=4.20, P<0.05) and less Barthel index score (OR=0.96, P<0.05) were independent risk factors for severe complications after PD in elderly patients. The risk prediction model of severe postoperative complications of PD was constructed using 6 indexes including age, hypertension, preoperative nutritional risk, operative type, Barthel index score in activities of daily living and operative time, and the fit of regression equation 0.93. ROC curve analysis showed that area under curve (AUC) was 0.76, the sensiti-vity 65%, and the specificity 76%. Conclusions Age 75 years or more, hypertension and low activities of daily living would be independent risk factors for severe complications after PD in elderly patients. The risk model constructed by age, hypertension, preoperative nutritional risk, operative type, Barthel index score in activities of daily living and operative time had certain predictive efficacy for severe complications after PD in elderly patients.

Objective To investigate the protective effect of Lycium barbanun glycopeptide(LbGP)on human keratinocyte HaCaT cells against radiation-induced cytotoxicity and its mechanism. Methods HaCaT cells were exposed to radiation with linear accelerator(rate 6 Gy/min)at doses of 4,8,12,16,20,24 and 28 Gy respectively,and the cell activity was detected by CCK-8 assay. HaCaT cells were treated with LbGP(0,0. 05,0. 1,0. 5,0. 8,1. 0,1. 5 and 3 mg/mL)for 4 h,exposed to radiation of 12 Gy,and detected for the cell viability by CCK-8 assay. HaCaT cells were divided into control group(without LbGP and radiation),radiation group(radiation of 12 Gy)and LbGP + radiation group(0. 8 mg/mL LbGP for 24 h,radiation of 12 Gy). After irradiation for 1 h,the reactive oxygen species(ROS)production was measured by flow cytometry,the superoxide dismutase(SOD)activity was determined by WST-8 assay and the expression of nuclear factor-E2 related factor 2(Nrf2),p-Nrf2,NADPH quinone oxidoreductase 1(NQO1)and heme oxygenase-1(HO-1)were detected by Western blot;The mRNA transcription levels of Nrf2,HO-1 and NQO1 were detected by qRT-PCR at 1,3 and 5 h after irradiation. Results Radiation of12 Gy induced about 50% cell death,and 0. 8 mg/mL LbGP showed the best protective effect on the activity of irradiated cells. After irradiation for 1 h,compared with the control group,the content of ROS in HaCaT cells increased significantly in radiation group(F = 2. 55,P < 0. 001),the activity of SOD decreased significantly(F = 1. 23,P < 0. 01),the contents of NQO1 and Nrf2 protein had no significant difference(F = 1. 78 and 1. 00,respectively,P > 0. 05),the content of HO-1protein increased significantly(F = 1. 37,P < 0. 05),and the content of p-Nrf2 protein decreased significantly(F = 2. 75,P < 0. 01);Compared with the radiation group,the content of ROS in HaCaT cells of LbGP + radiation group decreased significantly(F = 3. 61,P < 0. 001),the activity of SOD increased significantly(F = 1. 23,P < 0. 05),and the contents of Nrf2,p-Nrf2,HO-1 and NQO1 protein increased significantly(F = 4. 00,2. 25,6. 25 and 1. 27,respectively,P < 0. 05). In addition,the mRNA levels of Nrf2,HO-1 and NQO1 genes in LbGP + radiation group were significantly higher than those in radiation group at 1,3 and 5 h after irradiation(F = 0. 20～36. 00,P < 0. 05). Conclusion LbGP can mitigate oxidative stress damage of HaCaT cells induced by radiation and protect cell activity,which may play a role by activating Nrf2 and increasing the levels of its downstream antioxidant enzymes SOD,HO-1 and NQO1.

β-N-acetylglucosaminidases (NAGases) can convert natural substrates such as chitin or chitosan to N-acetyl-β-D glucosamine (GlcNAc) monomer that is wildly used in medicine and agriculture. In this study, the BcNagZ gene from Bacillus coagulans DMS1 was cloned and expressed in Escherichia coli. The recombinant protein was secreted into the fermentation supernatant and the expression amount reached 0.76 mg/mL. The molecular mass of purified enzyme was 61.3 kDa, and the specific activity was 5.918 U/mg. The optimal temperature and pH of the BcNagZ were 75 °C and 5.5, respectively, and remained more than 85% residual activity after 30 min at 65 °C. The Mie constant Km was 0.23 mmol/L and the Vmax was 0.043 1 mmol/(L·min). The recombinant BcNagZ could hydrolyze colloidal chitin to obtain trace amounts of GlcNAc, and hydrolyze disaccharides to monosaccharide. Combining with the reported exochitinase AMcase, BcNagZ could produce GlcNAc from hydrolysis of colloidal chitin with a yield over 86.93%.
Acetylglucosamine ; Acetylglucosaminidase ; Bacillus coagulans ; Chitin ; Chitinases ; Hydrogen-Ion Concentration ; Recombinant Proteins/genetics*

Objective: To investigate the effect of adding glutathion(GSH) to tumescent solution on autologous fat graft survival. Methods: 14 male and female New Zealand rabbits were divided into experimental group and control group randomly, 7 in each group. Experimental group: The donor areas of the rabbits were injected with 3 ml of tumescent solution with GSH. Control group: The donor areas of the rabbits were injected with 3 ml regular tumescent solution. DCFH-DA probe was used for fluorescent staining of harvested fat cells. Then stained fat cells were measured for the intracellular reactive oxygen species(ROS)content by fluorescence microplate. The grafts were harvested at 3 months after transplantation and assessed by general observation, volume measurement, wet weight measurement, HE staining for the number of fat cells, and CD34 immunohistochemical staining for the measurement of micro-vascular density. T test was performed by using SPSS 24.0. Results: The intracellular ROS content of harvested fat cells in experimental group was lower than that in control group, and the difference was statistically significant (P<0.05). At 3 months after transplantation, the wet weight of fat grafts in experimental group was (1133.21±87.97) mg and that in control group was (718.79±79.27) mg. The volume of fat grafts in experimental group was (1.00±0.04) ml and that in control group was (0.59±0.03) ml. The number of fat cells in experimental group was (746.6±15.7)/10 high magnification vision and that in control group was (350.1±32.4)/10 high magnification vision. The density in group experimental was (8.1±2.0)/high magnification vision and that in control group was (6.7±2.4)/high magnification vision. The grafts′ volume, wet weight, and the number of fat cells in experimental group were higher than those in control group, and the differences were statistically significant (P<0.05). The difference of micro-vascular density between experimental group and control group had no statistical significance (P>0.05). Conclusions The addition of GSH to tumescent solution optimizes the process of autologous fat harvest, thereby improving the survival of fat graft.