Objective To explore the potential role and underlying mechanism of the functionally uncharacterized gene Gm49394 on regulating β-cell apoptosis under diabetic conditions.Methods The expression and translational activity of Gm49394 in pancreatic β-cell lines and non-β-cell lines were validated using RNA fluorescence in situ hybridization(RNA-FISH),quantitative real-time PCR(qPCR),Western blotting,and immunofluorescence(IF)assay.The β-cell lines(NIT-1/Min6)with Gm49394 overexpression or knockdown were constructed.The proliferation,apoptosis,mitochondrial function,as well as oxidative stress and endoplasmic reticulum stress markers in these β-cell lines under physiological homeostasis or pathological stress conditions,such as high glucose(30 mmol/L),inflammation(10 ng/mL IFN-γ alone or combined with 10 ng/mL IL-6),and hydrogen peroxide(100 μmol/L H2O2)were detected by flow cytometry and Western blotting.Results RNA-FISH and qPCR indicated that Gm49394 was specifically expressed in pancreatic β-cell lines and up-regulated under high glucose or inflammatory stimulation.IF assay and Western blotting showed that Gm49394 had protein-coding activity.Flow cytometry and Western blotting identified that Gm49394 overexpression did not affect β-cell proliferation,but promoted β-cell apoptosis and increased reactive oxygen species(ROS)and mitochondrial superoxide(MitoSOX)levels in β cells under physiological homeostasis or pathological stress conditions(P<0.05).Under physiological conditions,Gm49394 knockdown failed to induce significant alterations on β-cell apoptosis,ROS,or MitoSOX levels.Under pathological stress conditions,Gm49394 knockdown significantly suppressed β-cell proliferation,apoptosis,as well as oxidative and endoplasmic reticulum stress(P<0.05).Conclusion Gm49394 may promote β-cell apoptosis via oxidative stress and endoplasmic reticulum stress.

ObjectiveThis study aims to investigate the effects of Linggui Zhugantang （LGZGT） on ventricular remodeling （VR） in mice with myocardial infarction （MI） and its impact on the Ras homologgene A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） signaling pathway. MethodsThe MI model of mice was established by ligating the left anterior descending coronary artery （LAD）. They were divided into the sham-operated group， the model group， the low-dose， medium-dose， and high-dose groups of LGZGT （2.34， 4.68， 9.36 g·kg^-1）， and the captopril group （3.25 mg·kg^-1）， with 10 mice in each group. After four weeks of continuous drug administration by gavage， the level of cardiac function in each group of mice was examined using small animal Doppler ultrasound. Hematoxylin-eosin （HE） staining and Masson staining was used to assess the morphological changes of myocardial tissue and calculate the rate of collagen fiber deposition in mouse myocardial tissue. Wheat germ agglutinin （WGA） staining was employed to compare the cross-sectional area of cardiomyocytes in each group of mice. The expression levels of α-smooth muscle actin （α-SMA）， matrix metalloproteinase-2 （MMP-2）， type Ⅰcollagen （Col Ⅰ）， Col Ⅲ， tissue inhibitor of metalloproteinase 1（TIMP1）， B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， Bcl-2， Caspase-3， and cleaved Caspase-3 were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to evaluate the mRNA levels of the pathway-related genes RhoA， ROCK1， and ROCK2. The protein expression levels of RhoA， ROCK1， and ROCK2 were tested by Western blot. ResultsThe level of cardiac function was markedly declined in the model group compared to the sham-operated group（P<0.01）. Myocardial tissue morphology changed significantly. The cross-sectional area of cardiomyocytes was significantly enlarged. The expression of α-SMA， MMP-2， Col Ⅰ， and Col Ⅲ was significantly upregulated（P<0.01）， and TIMP1 protein expression was significantly reduced（P<0.01）. The expressions of apoptosis-related proteins Bax were significantly up-regulated（P<0.01）， while the expression of Bcl-2 protein was significantly decreased（P<0.01）. The mRNA expression of RhoA， ROCK1， and ROCK2 were significantly upregulated （P<0.01）. Compared to the model group， the low-dose， medium-dose， and high-dose groups of LGZGT and the captopril group significantly reversed the experimental results of the model group in a dose-dependent manner （P<0.05， P<0.01）. ConclusionLGZGT significantly attenuated myocardial fibrosis， myocardial hypertrophy， and cardiomyocyte apoptosis after MI in mice and effectively reversed VR， the mechanism of which may be related to the modulation of the RhoA/ROCK signaling pathway.

ObjectiveThis study aims to investigate the effects of Linggui Zhugantang （LGZGT） on ventricular remodeling （VR） in mice with myocardial infarction （MI） and its impact on the Ras homologgene A （RhoA）/Rho-associated coiled-coil forming protein kinase （ROCK） signaling pathway. MethodsThe MI model of mice was established by ligating the left anterior descending coronary artery （LAD）. They were divided into the sham-operated group， the model group， the low-dose， medium-dose， and high-dose groups of LGZGT （2.34， 4.68， 9.36 g·kg^-1）， and the captopril group （3.25 mg·kg^-1）， with 10 mice in each group. After four weeks of continuous drug administration by gavage， the level of cardiac function in each group of mice was examined using small animal Doppler ultrasound. Hematoxylin-eosin （HE） staining and Masson staining was used to assess the morphological changes of myocardial tissue and calculate the rate of collagen fiber deposition in mouse myocardial tissue. Wheat germ agglutinin （WGA） staining was employed to compare the cross-sectional area of cardiomyocytes in each group of mice. The expression levels of α-smooth muscle actin （α-SMA）， matrix metalloproteinase-2 （MMP-2）， type Ⅰcollagen （Col Ⅰ）， Col Ⅲ， tissue inhibitor of metalloproteinase 1（TIMP1）， B-cell lymphoma-2 （Bcl-2）-associated X protein （Bax）， Bcl-2， Caspase-3， and cleaved Caspase-3 were detected by Western blot. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was used to evaluate the mRNA levels of the pathway-related genes RhoA， ROCK1， and ROCK2. The protein expression levels of RhoA， ROCK1， and ROCK2 were tested by Western blot. ResultsThe level of cardiac function was markedly declined in the model group compared to the sham-operated group（P<0.01）. Myocardial tissue morphology changed significantly. The cross-sectional area of cardiomyocytes was significantly enlarged. The expression of α-SMA， MMP-2， Col Ⅰ， and Col Ⅲ was significantly upregulated（P<0.01）， and TIMP1 protein expression was significantly reduced（P<0.01）. The expressions of apoptosis-related proteins Bax were significantly up-regulated（P<0.01）， while the expression of Bcl-2 protein was significantly decreased（P<0.01）. The mRNA expression of RhoA， ROCK1， and ROCK2 were significantly upregulated （P<0.01）. Compared to the model group， the low-dose， medium-dose， and high-dose groups of LGZGT and the captopril group significantly reversed the experimental results of the model group in a dose-dependent manner （P<0.05， P<0.01）. ConclusionLGZGT significantly attenuated myocardial fibrosis， myocardial hypertrophy， and cardiomyocyte apoptosis after MI in mice and effectively reversed VR， the mechanism of which may be related to the modulation of the RhoA/ROCK signaling pathway.

Objective:To observe the pregnancy outcomes of patients with unexplained recurrent spontaneous abortion (URSA) after interventional treatment or expectant treatment.Methods:This prospective study followed up 398 patients with recurrent spontaneous abortion from March 2017 to September 2022 in seven hospitals. Among them, 267 patients were diagnosed with URSA, including 124 patients who were initially diagnosed in the interventional treatment hospital and 143 patients who were initially diagnosed in the expectant treatment hospital. All URSA patients were followed up for 33 months. Ongoing pregnancy rates were observed as main outcome indicators.Results:A total of 127 patients became pregnant, and 107 of them had sustained pregnancies, the ongoing pregnancy rate was 84.25% (107/127). The ongoing pregnancy rate was 86.11% (31/36) in the interventional treatment group and 83.52% (76/91) in the expectant treatment group, with no significant difference ( P>0.05). During the follow-up, the ongoing pregnancy rates in the interventional treatment hospital and the expectant treatment hospital were 75.71% (53/70) and 94.74% (54/57), respectively, with a significant difference ( P<0.05). The ongoing pregnancy rate after interventional treatment in the interventional treatment hospital was 82.76% (24/29), which was similar to the 94.00% (47/50) after expectant treatment in the expectant treatment hospital ( P>0.05). Conclusion:The ongoing pregnancy rate of interventional treatment for URSA patients has not been significantly improved, suggesting that it may not be necessary to carry out this treatment.

Objective To investigate the three-dimensional changes in hard and soft tissues before and after treatment with digitally fabricated maxillary skeletal expanders in patients with maxillary transverse deficiency.Methods Twenty late adolescents or adults with maxillary transverse deficiency treated at the Orthodontic Department of Affiliated Stomatological Hospital of Nanjing Medical Uni-versity using digitally fabricated maxillary skeletal expanders were included.Cone-beam computed tomography(CBCT)scans were ob-tained before and after treatment.Three-dimensional measurements were analyzed using Dolphin 3D 11.95 software,and statistical anal-ysis was performed with SPSS software.Paired t-tests were used to compare pre-treatment and post-treatment changes in hard and soft tissues.Results Significant increases were observed in anterior midpalatal suture width,posterior midpalatal suture width,maxillary apical base width,and maxillary alveolar width(P＜0.001).No significant changes were found in alveolar ridge inclination,alveolar ridge height,LSNA(P＞0.05).Significant increases occurred in apical distance,cementoenamel junction distance,and inter-fossa distance(P＜0.001),while first molar inclinationand ∠U1-SN remained stable(P＞0.05).Significant improvements were noted in upper airway volume,nasal floor base width,and nasal cavity width(P＜0.001).Conclusion For late adolescents or adults with max-illary transverse deficiency,digitally fabricated maxillary skeletal expanders can achieve near-parallel expansion of the midpalatal suture,effectively improving maxillary transverse dimensions and airway volume while minimizing dental side effects.

Objective:To observe the pregnancy outcomes of patients with unexplained recurrent spontaneous abortion (URSA) after interventional treatment or expectant treatment.Methods:This prospective study followed up 398 patients with recurrent spontaneous abortion from March 2017 to September 2022 in seven hospitals. Among them, 267 patients were diagnosed with URSA, including 124 patients who were initially diagnosed in the interventional treatment hospital and 143 patients who were initially diagnosed in the expectant treatment hospital. All URSA patients were followed up for 33 months. Ongoing pregnancy rates were observed as main outcome indicators.Results:A total of 127 patients became pregnant, and 107 of them had sustained pregnancies, the ongoing pregnancy rate was 84.25% (107/127). The ongoing pregnancy rate was 86.11% (31/36) in the interventional treatment group and 83.52% (76/91) in the expectant treatment group, with no significant difference ( P>0.05). During the follow-up, the ongoing pregnancy rates in the interventional treatment hospital and the expectant treatment hospital were 75.71% (53/70) and 94.74% (54/57), respectively, with a significant difference ( P<0.05). The ongoing pregnancy rate after interventional treatment in the interventional treatment hospital was 82.76% (24/29), which was similar to the 94.00% (47/50) after expectant treatment in the expectant treatment hospital ( P>0.05). Conclusion:The ongoing pregnancy rate of interventional treatment for URSA patients has not been significantly improved, suggesting that it may not be necessary to carry out this treatment.

Objective To investigate the three-dimensional changes in hard and soft tissues before and after treatment with digitally fabricated maxillary skeletal expanders in patients with maxillary transverse deficiency.Methods Twenty late adolescents or adults with maxillary transverse deficiency treated at the Orthodontic Department of Affiliated Stomatological Hospital of Nanjing Medical Uni-versity using digitally fabricated maxillary skeletal expanders were included.Cone-beam computed tomography(CBCT)scans were ob-tained before and after treatment.Three-dimensional measurements were analyzed using Dolphin 3D 11.95 software,and statistical anal-ysis was performed with SPSS software.Paired t-tests were used to compare pre-treatment and post-treatment changes in hard and soft tissues.Results Significant increases were observed in anterior midpalatal suture width,posterior midpalatal suture width,maxillary apical base width,and maxillary alveolar width(P＜0.001).No significant changes were found in alveolar ridge inclination,alveolar ridge height,LSNA(P＞0.05).Significant increases occurred in apical distance,cementoenamel junction distance,and inter-fossa distance(P＜0.001),while first molar inclinationand ∠U1-SN remained stable(P＞0.05).Significant improvements were noted in upper airway volume,nasal floor base width,and nasal cavity width(P＜0.001).Conclusion For late adolescents or adults with max-illary transverse deficiency,digitally fabricated maxillary skeletal expanders can achieve near-parallel expansion of the midpalatal suture,effectively improving maxillary transverse dimensions and airway volume while minimizing dental side effects.

Objective: To analyze the disease burden and risk factors of colorectal cancer in Zhejiang Province from 1990 to 2019, so as to provide the basis for prevention and control of colorectal cancer. Methods: Based on data of 2019 Global Burden of Disease (GDB 2019), disease burden and risk factors of colorectal cancer in Zhejiang Province from 1990 to 2019 was assessed using years of life lost (YLL), years lived with disability (YLD), disability-adjusted life years (DALY). Results: In 2019, the YLL rate, YLD rate and DALY rate caused by colorectal cancer in Zhejiang Province were 496.15/105, 31.81/105 and 527.96/105, respectively. From 1990 to 2019, the YLL rate, YLD rate and DALY rate caused by colorectal cancer in Zhejiang Province increased by 114.90%, 482.60% and 123.38%, respectively, showing increasing trends (average annual percent change values were =2.663, 6.283 and 2.800, respectively,all P<0.05). From 1990 to 2019, the YLL rate, YLD rate and DALY rate in the age groups of 15 to 49 years, 50 to 69 years and 70 years and older showed increasing trends (all P<0.05). In 1990, the top ten risk factors for colorectal cancer in Zhejiang Province were diet low in calcium, diet low in milk, diet low in whole grains, smoking, alcohol use, low physical activity, high fasting plasma glucose, diet high in red meat, diet low in fiber and high body mass index. In 2019, the top ten risk factors for colorectal cancer in Zhejiang Province were diet low in milk, diet low in whole grains, diet low in calcium, alcohol use, diet high in red meat, high body mass index, high fasting plasma glucose, low physical activity, diet low in fiber and diet high in processed meat. Conclusions The disease burden of colorectal cancer in Zhejiang Province showed an upward trend from 1990 to 2019. The top ten risk factors for colorectal cancer remained between 1990 and 2019, while there was a slight change in ranking.

Objective To compare and analyze the differences in CD8+naive,effector,memory,exhausted and regulatory T cells in order to investigate the impact of gender on the differentiation fate of CD8+T cells in the context of type 1 diabetes (T1D)based on female and male non-obese diabetic (NOD)mice and healthy Institute for Cancer Research (ICR)mice.Methods The frequencies and phenotypes of CD8+T cell differentiation subsets including naive T cells (TN),central memory T cells (TCM),effector T cells(TEFF),effector precursor T cells (TEP),exhausted T cells (TEX),precursor exhausted T cells (TPEX)and regulatory T cells (Tregs)in the spleen,pancreatic draining lymph nodes (pLN)and pancreas infiltrating lymphocytes (PIL)of male and female NOD mice were detected by flow cytometry.Results The frequencies of IFN-γ+,CD107a+and CCL5+CD8+TEFF in pLN and PIL of female NOD mice were significantly higher than those of male NOD mice.However,the frequencies of CD8+TN,CD8+TCM,CD8+TEX,CD8+TPEX and CD122+CD8+Tregs subsets in the spleen were significantly decreased.While there were no significant differences in the above CD8+T cell subsets except CD8+Tregs between female and male ICR mice. Conclusion Androgen may inhibit the differentiation of memory T cells into effector T cells and promote the exhaustion of effector T cells,leading to the difference in morbidity between the male and female mice.

Objective This study aims to investigate the therapeutic effect and mechanism of Naoxinqing on non-alcohol fatty liver disease (NAFLD) induced by a high-fat diet through network pharmacology,molecular docking and in vitro and in vivo experiments. Methods ApoE-/-mice were given a high-fat diet for 12 weeks to establish the NAFLD model,followed by a 12-week Naoxinqing administration. To evaluate the therapeutic effect of Naoxinqing on NAFLD induced by a high-fat diet,biochemical and histopathological experiments were performed,including assessment of blood lipids,liver function,serum inflammatory factors,as well as Hematoxylin and eosin (HE),Oil red O,and Sirius red staining of liver. Subsequently,network pharmacology and molecular docking techniques were employed to predict the key targets of Naoxinqing. Finally,the mechanism of Naoxinqing was validated by Western Blot in HepG2 cells and liver tissue. Results The results of serum biochemistry and liver tissue pathology showed that Naoxinqing can significantly improve high-fat diet-induced hepatic lipid accumulation,hepatocellular injury,and inflammation. Network pharmacology and molecular docking analysis results suggested that Naoxinqing may affect lipid metabolism through the AMP-activated protein kinase (AMPK)/Sirtuin 1 (SIRT1) pathway. Finally,in vitro cell experiment confirmed that the main mechanism of Naoxinqing is to activative the AMPK/SIRT1 pathway,upregulate the expression of downstream carnitine palmitoyltransferase 1 (CPT1A),promote fatty acid oxidation,and ultimately improve NAFLD. Conclusion This study demonstrated that Naoxinqing improved NAFLD by promoting fatty acid oxidation through the activation of the AMPK/SIRT1 pathway.