Objective:To separate and purify the polysaccharides from Polygonatum filipes,characterize their primary structure and investigate the immunomodulatory effects on RAW264.7 macrophages.Methods:Crude polysaccharides from Polygonatum filipes were extracted by ultrasound assisted method,then Polygonatum filipes polysaccharides(CSPFPs)were obtained after elimination of the proteins with combined papain-Sevag method.The total sugar content was determined by phenol-sulfuric acid method.Structures of CSPFPs were analyzed by fourier transform infrared spectroscopy(FT-IR),high performance gel permeation chromatography(HPGPC)and high performance liquid chromatography(HPLC).Effects of CSPFPs on cell viability,pinocytic activity,TNF-α secretion,MAPK and NF-κB signaling pathways of RAW264.7 cells were explored by MTT,Neutral red,ELISA and Western blot,respectively.Results:Extraction rate of CSPFPs by ultrasound-assisted method was 41.61%,which contained total sugar content of 94.00%.CSPFPs with Mw of 3 125 Da was composed of arabinose(1.85%),galactose(6.14%),glucose(56.41%)and mannose(35.60%).The in vitro experiments showed that CSPFPs were non-cytotoxic and enhanced the pinocytic activity,TNF-α secretion and phosphorylation levels of p38,ERK,JNK,p65,IκB and IKK,indicating the activation of MAPK and NF-κB signaling pathways under the concentra-tion of 2.5～200 μg/ml.Conclusion:The ultrasound-assisted method can efficiently isolate CSPFPs with immunomodulatory activity,which provides basic data for the development and application of CSPFPs as an immunostimulant.

This study analyzed the clinical and laboratory data of 3 children diagnosed with mitochondrial disease-associated primary adrenal insufficiency (PAI) at the Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from October 2018 to November 2023.All patients were normal at birth but gradually developed symptoms and were diagnosed with PAI between the ages of 1 year and 1 month and 7 years and 3 months.The children presented typical clinical symptoms of PAI, including skin and mucosal hyperpigmentation (3 cases), electrolyte disturbances (3 cases), and hypoglycemia (2 cases), as well as multisystem abnormalities related to mitochondrial disease, including recurrent infections, growth retardation, cachexia, and hyperlactatemia.Genetic testing revealed significant single deletions in mitochondrial DNA in all patients: Patient 1: m.11219_15954del, Patient 2: m.8483_13459del, and Patient 3: m.8649_16084del.Treatment with Hydrocortisone acetate replacement therapy improved the electrolyte disturbances and hypoglycemia, but issues with recurrent infections, growth retardation, and cachexia persisted.This study suggests that in clinical practice, the possibility of mitochondrial disease should be highly suspected when PAI patients present with multisystem abnormalities, especially in conjunction with hyperlactatemia.

To train applied stomatology students, a stomatology practice education system was created and practiced in conjunction with post competency in the context of New Medicine. The practice teaching system is featured by six progressive stages and integration of knowledge and practice, uses student-centered and ability-oriented teaching model throughout the education, integrates the curriculum ideology system of working together in the same direction and cultivating talents with high moral standards, and forms a multi-cooperative and people-oriented evaluation system. The stomatology practice education system led by the New Medicine with collaboration of multiple elements such as "medicine, teaching, research, and innovation" has contributed to the comprehensive improvement of post competency in stomatology students. This system has played an important supportive role in the training of applied stomatology talents in the new era.

To train applied stomatology students, a stomatology practice education system was created and practiced in conjunction with post competency in the context of New Medicine. The practice teaching system is featured by six progressive stages and integration of knowledge and practice, uses student-centered and ability-oriented teaching model throughout the education, integrates the curriculum ideology system of working together in the same direction and cultivating talents with high moral standards, and forms a multi-cooperative and people-oriented evaluation system. The stomatology practice education system led by the New Medicine with collaboration of multiple elements such as "medicine, teaching, research, and innovation" has contributed to the comprehensive improvement of post competency in stomatology students. This system has played an important supportive role in the training of applied stomatology talents in the new era.

This study analyzed the clinical and laboratory data of 3 children diagnosed with mitochondrial disease-associated primary adrenal insufficiency (PAI) at the Guangzhou Women and Children′s Medical Center, Guangzhou Medical University from October 2018 to November 2023.All patients were normal at birth but gradually developed symptoms and were diagnosed with PAI between the ages of 1 year and 1 month and 7 years and 3 months.The children presented typical clinical symptoms of PAI, including skin and mucosal hyperpigmentation (3 cases), electrolyte disturbances (3 cases), and hypoglycemia (2 cases), as well as multisystem abnormalities related to mitochondrial disease, including recurrent infections, growth retardation, cachexia, and hyperlactatemia.Genetic testing revealed significant single deletions in mitochondrial DNA in all patients: Patient 1: m.11219_15954del, Patient 2: m.8483_13459del, and Patient 3: m.8649_16084del.Treatment with Hydrocortisone acetate replacement therapy improved the electrolyte disturbances and hypoglycemia, but issues with recurrent infections, growth retardation, and cachexia persisted.This study suggests that in clinical practice, the possibility of mitochondrial disease should be highly suspected when PAI patients present with multisystem abnormalities, especially in conjunction with hyperlactatemia.

Objective:To separate and purify the polysaccharides from Polygonatum filipes,characterize their primary structure and investigate the immunomodulatory effects on RAW264.7 macrophages.Methods:Crude polysaccharides from Polygonatum filipes were extracted by ultrasound assisted method,then Polygonatum filipes polysaccharides(CSPFPs)were obtained after elimination of the proteins with combined papain-Sevag method.The total sugar content was determined by phenol-sulfuric acid method.Structures of CSPFPs were analyzed by fourier transform infrared spectroscopy(FT-IR),high performance gel permeation chromatography(HPGPC)and high performance liquid chromatography(HPLC).Effects of CSPFPs on cell viability,pinocytic activity,TNF-α secretion,MAPK and NF-κB signaling pathways of RAW264.7 cells were explored by MTT,Neutral red,ELISA and Western blot,respectively.Results:Extraction rate of CSPFPs by ultrasound-assisted method was 41.61%,which contained total sugar content of 94.00%.CSPFPs with Mw of 3 125 Da was composed of arabinose(1.85%),galactose(6.14%),glucose(56.41%)and mannose(35.60%).The in vitro experiments showed that CSPFPs were non-cytotoxic and enhanced the pinocytic activity,TNF-α secretion and phosphorylation levels of p38,ERK,JNK,p65,IκB and IKK,indicating the activation of MAPK and NF-κB signaling pathways under the concentra-tion of 2.5～200 μg/ml.Conclusion:The ultrasound-assisted method can efficiently isolate CSPFPs with immunomodulatory activity,which provides basic data for the development and application of CSPFPs as an immunostimulant.

Objective:Analyze the impact of smoking on the mortality and life expectancy of residents in Tianjin in 2019.Methods:Use mortality case-control study method to collect all cause of death cases of residents in Tianjin in 2019 for analysis. After adjusting for the 5-years-old age group, education level, and marital status, the smoking attributed deaths from different diseases of different genders, smoking attributed deaths in different age groups, and their impact on life expectancy were analyzed.Results:The total number of deaths in 2019 was 75 254, with 42 201 males (56.1%). Among male deaths, 3 215 (9.9%) were attributed to smoking, of which 2 157 (50.2%) lung cancer deaths were attributed to smoking; The risk of lung cancer death among smokers was 3.075 times higher than that of non-smokers (95% CI: 2.812-3.364); Among the 33 053 female deaths (43.9%), 1 396 (5.8%) were caused by smoking, with 744 (29.1%) lung cancer deaths attributed to smoking. The age group with the highest number of deaths attributed to smoking for women was the 75-<80 years old age group, followed by the 70-<75 and 80-<85 years old age groups. The age group with the highest proportion of deaths attributed to smoking for men was the 55-<60 years old age group. In addition, smoking accounts for more than 60% of deaths in the 60-<65, 45-<50, 55-<60, and 65-<70 years old age groups. In 2019, the loss of life expectancy attributed to smoking deaths among all residents in Tianjin was 1.13 years, with a loss of 1.15 years for males and 0.57 years for females. The expected life expectancy excluding deaths caused by smoking was 82.92 years, 80.77 years for males and 84.61 years for females. Conclusions:Smoking remains one of the important risk factors for death among residents. Promoting effective measures to reduce smoking rates is an effective way to increase life expectancy.

Objective To investigate the influence factors of lymphovascular space invasion(LVSI)in patients with low-grade endometrioid carcinoma.Methods Clinical data of 167 patients with low-grade(G1,G2)endometrioid carcinoma of uterus diagnosed by surgical pathology from January 2015 to May 2023 were retrospectively analyzed.Univariate analysis of the following 9 indicators was performed:age,postmenopausal status,abnormal vaginal bleeding duration,combined with metabolic syndrome,increased CA125(≥ 35 U/ml),endometrial thickness,uterine occupation,combined with adenomyosis of uterus,stage of the International Federation of Gynecology and Obstetrics(FIGO)2009 standards.The binary logistic refression analysis was made on factors with P＜0.05.Results Among the 167 patients with low-grade endometrioid carcinoma of uterus,24 cases(14.4％)had LVSI.The binary logistic regression analysis was performed for 4 factors(abnormal vaginal bleeding duration ≥ 4 months,increased CA125,combined with adenomyosis of uterus,stage Ⅱ and above)with P＜0.05 in the univariate analysis,and the results showed stage Ⅱ and above(OR=7.357,95％CI:2.140-25.288,P=0.002)and increased CA125(OR=4.883,95％CI:1.612-14.794,P=0.005)were an independent prognostic factor for LVSI in low-grade endometrial carcinoma of uterus.Conclusion Stage Ⅱ(FIGO 2009)and above and CA125≥35 U/ml are associated with LVSI in patients with low-grade endometrioid carcinoma,indirectly suggesting the risk of lymph node metastasis,which should be paid close attention to before surgery in order to develop a more accurate surgical plan.

Objective To screen the differentially expressed proteins in glomeruli during the treatment of lupus nephritis(LN)with hydroxychloroquine sulfate(HCQ).Methods Sixty female NZB/WF1 mice were used.At the age of 28 weeks,40 mice with proteinuria of 3+to 4+were divided into HCQ and control groups.After feeding for 36 weeks,the glomerular proteins were extracted by magnetic beads and analyzed by two-dimensional fluorescence difference gel electrophoresis(2D-DIGE)and matrix-assisted laser desorption/ioniza-tion time-of-flight mass spectrometry(MALDI-TOF-MS).The expression of the candidate proteins in the glomeruli of the LN mice was exa-mined by immunohistochemistry.Results A total of 31 differentially expressed proteins were identified between the two groups,including 24 upregulated and seven downregulated proteins.Conclusion The expression of proteins like RI and ENOA in the glomeruli of LN mice was significantly different during HCQ treatment.These proteins may be involved in the pathogenesis of LN and are therefore potential targets of HCQ in the treatment of LN.

Objective To establish an improved simple and efficient method for isolation and culture of primary lung fibroblasts(LFB)from neonatal mice,and study their growth characteristics in vitro.Methods The lungs of 3-days-old mice or 8-weeks-old mice were taken under sterile conditions,and the stromal tissue was cut into 1mm3 tissue mass.High-glucose DMEM culture medium containing 10g/dl fetal bovine serum(FBS)or 20g/dl FBS was used for tissue adhesion culture.The LFB was purified by the differential time adhesion method,and the growth morphology and adherence state of the cells were observed dynamically under an inverted microscope.The primary LFB was identified by flow cytometry.The activity of the third-generation cell after culture was detected by CCK-8 assay.Results Lung tissues in mice at 3 days of age and 20g/dl FBS concentration cultured by the improved tissue mass adherent cultured method began to grow radially to the periphery on the 2nd day.On the day 7th,the cells growth density reached 90%,and the cell morphology was as a spindle.The CD140a positive and CD45 negative cells reached more than 90%after purification by differential time adhesion method,and the cells still maintain good cell activity after cultured for 3 generations.Conclusion The improved tissue adhesion method can obtain a large number of purified mice LFB with good activity simply and efficiently,which lays a foundation for the study of lung inflammation,tumors and in vitro efficacy of drugs.