ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

ObjectiveTo investigate the effects of different fermentation methods and times on the fungal flora and chemical composition of Aurantii Fructus， in order to obtain the optimal fermentation conditions and flora structure， and to ensure the stability and controllability of the fermented varieties. MethodsScanning electron microscopy was used to observe and analyze the colony characteristics on the surface of Aurantii Fructus under different fermentation conditions. Internal transcribed spacer 2（ITS2） high-throughput sequencing， combined with fungal community diversity analysis and fungal community structure analysis， were used to obtain the fungal flora microbial categories of Aurantii Fructus under the conditions of traditional pressure-shelf fermentation and non-pressure-shelf natural fermentation for 7， 14， 21 d（numbered Y1-Y3 for the former， and numbered F1-F3 for the latter）， respectively. At the same time， the chemical components in the fermentation process were detected by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS/MS）， combined with principal component analysis（PCA）， partial least squares-discriminant analysis（PLS-DA） and compound retention time， parent ions， characteristic fragment ions and other information， the differential compounds between the different fermentation samples were screened and identified. ResultsThe analysis of fungal community diversity showed that the dominant flora did not change at different fermentation time points in the traditional pressure-shelf fermentation method， while in the non-pressure-shelf natural fermentation method， there was a significant difference with the fermentation process， and at the genus level， the dominant genus of samples Y1， Y2， Y3 and F2 was Aspergillus， while the dominant genera of samples F1 and F3 were both Rhizopus. This indicated that the microbial growth environment provided by the traditional fermentation method was more stable， and the microbial community structure was more stable， which was more conducive to the stable and controllable fermentation process and fermented products. A total of 155 compounds were identified by compositional analysis， including 70 flavonoids， 38 coumarins， 10 alkaloids， 34 organic acids and 3 other compounds. After fermentation， two new components of ribalinine and pranferin were produced. Different fermentation conditions also brought about differences in chemical composition， multivariate statistical analysis obtained 26 differential compounds under two different fermentation methods， mainly including flavonoids， organic acids and coumarins. Comprehensively， the microbial community structure of samples fermented by the traditional pressure-shelf method of Aurantii Fructus for 14 d was stable， the species richness was high and the overall content of differential compounds was high， which was the optimal processing condition. ConclusionCompared with non-pressure-shelf natural fermentation， the traditional method has obvious advantages in terms of the stability of the microbial community structure and the content of chemical compounds， and the optimal condition is 14 days of fermentation. This study is helpful to promote the quality stability and fermentation bioavailability of fermented products of Aurantii Fructus， as well as to provide an experimental basis for the further improvement of the quality control methods of this variety.

Objective To explore the effect of intelligent rehabilitation exercise device on improving postoperative self-efficacy of breast cancer patients,promoting postoperative functional recovery of affected limbs and improving postoperative upper limb edema.Methods A total of 60 postoperative breast cancer patients in Jiaxing Hospital of Traditional Chinese Medicine were selected from January 2022 to Apil 2023,all of them underwent axillary dissection.The patients were randomly divided into control group(30 cases)and experimental group(30 cases)according to random number method.The patients in control group were treated with postoperative upper limb functional exercise,and the patients in experimental group were treated with intelligent rehabilitation exercise device after breast cancer surgery.Self-efficacy and functional exercise compliance at 3 months after operation,shoulder range of motion and incidence of upper limb edema 1 month and 3 months after operation were compared between two groups.Results After 3 months of surgery,the self-efficacy and functional exercise compliance of patients in experimental group were higher than those in control group(P＜0.05);After surgery 1 month and 3 months,the shoulder joint range of motion of patients in experimental group was better than that in control group(P＜0.05);The incidence of postoperative upper limb edema in experimental group was lower than that in control group 1 month after surgery,and the difference was not statistically significant(P＞0.05).The experimental group was lower than control group 3 months after surgery,and the difference was statistically significant(P＜0.05).Conclusion Intelligent rehabilitation exercise device after breast cancer operation can improve patients'self-efficacy,improve patients'compliance with functional exercise,promote functional recovery of affected limbs,prevent and treat postoperative upper limb edema,which is worthy of clinical promotion.

Objective: To investigate the attitude and behaviors towards tobacco control among civil servants in smoke-free governments of Ningbo City, so as to provide insights into the construction of smoke-free governments. Methods: Ten governments were randomly sampled from city-level and county/district-level smoke-free governments in Ningbo City using a stratified random sampling method, and 10 civil servants were sampled from each government using a convenient sampling method. Participants' demographics, attitude and behaviors towards tobacco control and second-hand smoke exposure were collected using a self-designed questionnaire and descriptively analyzed. Results: A total of 1 100 civil servants were recruited, including 466 men (42.36%) and had a mean age of (39.02±9.38) years. There were 1 005 participants favoring “civil servants should refuse smoking firstly” (91.36%), 1 058 participants favoring “governments should build a smoke-free environment firstly” (96.18%), 1 075 participants favoring “smoking should be completely banned in any indoor public places” (97.73%), 913 participants thinking that smoking should be completely banned in any indoor public places (83.00%) and 813 participants thinking that tobacco control regulations are implemented satisfactorily in their workplaces (73.91%). The overall prevalence of current smoking was 11.91% among participants, and the prevalence of current smoking was 28.11% among male participants. Among all current smokers, 84 attempted to quit smoking (64.12%), and 89 had willingness or plans to quit smoking (67.94%). The proportion of second-hand smoke exposure in workplaces was 40.97% among non-smokers in the past one week. Conclusions The civil servants in Ningbo City has correct awareness of smoke-free governments, active attitudes towards tobacco control and low prevalence of smoking. However, smoke-free regulations remain to be improved and long-term tobacco control mechanisms remain to be created in governments in Ningbo City

Objective: To investigate the e-cigarette use and its influencing factors among middle school students in Ningbo City, Zhejiang Province, so as to provide insights into e-cigarette control among adolescents. Methods: Students of junior high schools, high schools and vocational high schools in Ningbo City were sampled using a multi-stage stratified cluster random sampling method from September to October 2021. Students' general information, cigarette and e-cigarette use and e-cigarette advertising exposure were collected using 2021 China Youth Tobacco Survey Questionnaire, and proportions of e-cigarette current use and attempt to use were analyzed. Factors affecting the attempt to use e-cigarettes were identified using a multivariable logistic regression model. Results: Totally 1 591 questionnaires were allocated, and 1 574 valid questionnaires were recovered, with an effective rate of 98.93%. The respondents included 863 males (54.83%) and 711 females (45.17%); 787 junior high school students (50.00%), 535 high school students (33.99%) and 252 vocational high school students (16.01%). The proportion of current e-cigarette use was 1.91% and the proportion of attempt to use e-cigarettes was 6.61%. Multivariable logistic regression analysis identified males (OR=2.544, 95%CI: 1.438-4.501), living in rural areas (OR=2.084, 95%CI: 1.195-3.634), having close friends who were smokers (OR=4.828, 95%CI: 2.936-7.940), witnessing teachers smoking at school (OR=2.766, 95%CI: 1.642-4.659), e-cigarette advertising exposure (OR=2.155, 95%CI: 1.315-3.532), current cigarette use (OR=9.365, 95%CI: 2.610-33.603) and attempt to use cigarettes (OR=7.519, 95%CI: 4.115-13.737) as promoting factors for the attempt to use e-cigarette. Conclusions There are e-cigarette use behaviors among middle school students in Ningbo City. Having teachers or close friends who were smokers, cigarette use behaviors and e-cigarette advertising exposure may increase the risk of attempt to use e-cigarettes among middle school students.

Objective: To investigate the prevalence of diabetes among individuals at high risk of diabetes in Baoshan District, Shanghai Municipality, so as to provide insights into community-based diabetes management. Methods: Permanent residents at ages of 35 years and older were sampled from Baoshan District using a multistage stratified cluster sampling method, and residents at a high risk of diabetes were screened using the Form for Risk Assessment of Diabetes among Community Residents in Shanghai Municipality. Participants' demographics, disease history and history of medication were collected using questionnaire surveys, and height, body weight, waist circumference, hip circumference, and blood pressure were measured. Diabetes was screened using fasting blood glucose and glucose tolerance test. The factors affecting the development of diabetes were identified among high-risk residents for diabetes using a multivariable logistic regression model. Results: A total of 3 107 residents at a high risk for diabetes were enrolled, including 1 165 men (37.50%) and 1 942 women (62.50%) with a mean age of (63.58±9.77) years. The prevalence of diabetes was 21.69% among the study subjects, and multivariable logistic regression analysis showed that men (OR=1.689, 95%CI: 1.357-2.104), age (40 years-, OR=4.833, 95%CI: 1.036-22.553; 50 years-, OR=2.627, 95%CI: 1.432-4.819; 60 years-, OR=1.551, 95%CI: 1.119-2.150; 70 years and older, OR=1.579, 95%CI: 1.232-2.025); high school/technical secondary school (OR=2.677, 95%CI: 1.636-4.380), overweight/obesity (OR=1.891, 95%CI: 1.447-2.472), hypertension (OR=1.306, 95%CI: 1.049-1.626), dyslipidemia (OR=1.428, 95%CI: 1.114-1.831), history of impaired glucose regulation (OR=15.161, 95%CI: 11.827-19.434) and family history of type 2 diabetes mellitus (OR=2.092, 95%CI: 1.619-2.704) caused an increased risk of diabetes among residents at a high risk diabetes. Conclusions The prevalence of diabetes was 21.69% among high-risk populations of diabetes in Baoshan District. Gender, age, educational level, overweight/obesity, hypertension, dyslipidemia, history of impaired glucose regulation and family history of type 2 diabetes mellitus are factors affecting the development of diabetes among high-risk populations.

Objective:To explore the predictive value of serum 25-hydroxyvitamin D [25 (OH) D] and interleukin-6 (IL-6) levels in the development of osteoporosis (OP) in gestational diabetes mellitus (GDM) during early pregnancy.Methods:A total of 178 pregnant women with GDM admitted to Shanxi Provincial Children’s Hospital from Sep. 2018 to May. 2022 were selected as the research subjects and divided into OP group and non-OP group based on bone density. General information of patients was collected, and biochemical indicators related to bone metabolism were measured, including calcium (Ca), β-collagen special sequence ( β-CTX), type 1 procollagen amino-terminal peptide (PINP), n-terminal bone calciuim (N-MID), osteocalcin (OC), and bone alkaline phosphatase (BAP). The levels of 25 (OH) D and IL-6 in the serum of the two groups were measured. The predictive value of 25 (OH) D and IL-6 levels for GDM complicated with OP was analyzed. Results:Compared with the non-OP group (29.95±6.88), the serum 25 (OH) D level was significantly lower in the OP group (20.78±6.02) ( t=5.56, P<0.001), and the IL-6 level was significantly higher in the OP group (75.19±9.15) than in the non-OP group (50.23±10.08) ( t=10.26, P<0.001). There was no significant difference in the levels of Ca, P, β-CTX, PINP, or N-MID between the two groups (all P>0.05). The levels of BAP (13.25±3.12) and OC (6.62±1.24) in the OP group were significantly lower than those in the non-OP group (BAP: 18.23±3.78, t=5.52, P<0.001; OC: 8.75±1.52, t=5.87, P<0.001). Pearson correlation analysis showed a positive correlation between 25 (OH) D level and bone density, BAP, and OC ( r=0.26, 0.21, 0.23, all P<0.05), while IL-6 level showed a negative correlation with bone density, BAP, and OC ( r=-0.39, -0.21, -0.27, all P<0.05). ROC curve analysis showed that serum 25 (OH) D and IL-6 could serve as effective diagnostic markers for GDM complicated with OP (AUC=0.853, P<0.0001; AUC=0.869, P<0.0001) during early pregnancy. Conclusions:The serum 25 (OH) D level is decreased and IL-6 level is increased in GDM patients with OP during early pregnancy. The levels of serum 25 (OH) D and IL-6 have predictive value for the development of GDM complicated with OP during early pregnancy.

Objective: To investigate the prevalence of intention to smoke and current smoking and identify its influencing factors among middle school students in Ningbo City, so as to provide insights into the development and implementation of tobacco control among adolescents. Methods: The epidemiological data of tobacco use among adolescents in Ningbo City in 2021 were collected, including demographics, tobacco use, parents' and close friends' smoking status, secondhand smoke exposure and tobacco advertisement exposure. The intention to smoke, current smoking, secondhand smoke exposure and tobacco advertisement exposure were descriptively analyzed among middle school students, and the factors affecting intention to smoke, current smoking were identified using a multivariable logistic regression analysis. Results: Totally 1 591 questionnaires were allocated to middle school students, and 1 574 valid questionnaires were recovered, with an effective recovery rate of 98.93%. The respondents included 787 junior high school students, 535 high school students and 252 vocational high school students, and the prevalence was intention to smoke and current smoking was 6.73% and 1.78% among the respondents, respectively. Multivariable logistic regression analysis identified boys (OR=2.398, 95%CI: 1.466-3.922), high school students (OR=2.051, 95%CI: 1.212-3.470), vocational high school students (OR=2.011, 95%CI: 1.108-3.650), more than 100 yuan disposable pocket money per week (OR=2.534, 95%CI: 1.301-4.935) and close friends' smoking (OR=4.310, 95% CI: 2.755-6.757) as risk factors of intention to smoke, and boys (OR=12.420, 95%CI: 1.645-93.753), close friends' smoking (OR=11.628, 95%CI: 4.348-31.250) and parents' smoking (OR=2.667, 95%CI: 1.029-6.897) as risk factors of current smoking among middle school students. The prevalence of secondhand smoke exposure and tobacco advertisement exposure was 76.43% and 51.65%, and television/video/cinema was the most likely route of tobacco advertisement exposure (42.06%). Conclusions The prevalence of intention to smoke and current smoking is low among middle school students in Ningbo City; however, the proportions of exposure to secondhand smoke and tobacco advertisement are relatively high. Tobacco control interventions are recommended to be intensified targeting high-risk populations, such as vocational high school students.

Objective:To explore the anti-inflammatory effect of pomegranate peel polyphenols on a rat auriclular model of acne and its mechanism of action.Methods:Totally, 36 specific-pathogen-free SD rats were randomly divided into 6 groups: blank group, model group, low-, medium- and high-dose pomegranate peel polyphenol groups and positive control group. In all groups except the blank group, 0.5 ml of 100% oleic acid was applied to the openings of bilateral auricular ducts once a day for 3 consecutive weeks, followed by subcutaneous injections of 50 μl of Propionibacterium acnes suspension at the oleic acid-applied sites once a day for 3 consecutive days, so as to establish a rat auriclular model of acne. After the model was confirmed to be successfully established by naked eyes, the low-, medium-, high-dose pomegranate peel polyphenol groups were topically treated with 0.5 mg of 1.4%, 2.8%, 5.6% (mass fraction) pomegranate peel polyphenol ointment respectively, the positive control group was topically treated with 0.5 mg of clindamycin hydrochloride gel, and the blank group and model group were topically treated with the same amount of distilled water. All the topical treatments were performed twice a day for 2 consecutive weeks. Twenty-four hours after the last topical treatment, abdominal aortic blood samples were collected, and enzyme-linked immunosorbent assay (ELISA) was conducted to detect the serum level of interleukin 17 (IL-17) in rats; rat auricular tissues were resected, hematoxylin-eosin (HE) staining was performed to observe histopathological changes of the skin tissues in each group, and immunohistochemical study to determine the expression of mammalian target of rapamycin (mTOR) , hypoxia-inducible factor-1α (HIF-1α) , and retinoic acid-related orphan receptor-γt (RORγt) in local tissues. Data meeting the assumptions of homogeneity of variances were analyzed by using one-way analysis of variance, and those that did not meet the assumptions of homogeneity of variances were analyzed by using Kruskal-Wallis H test; multiple comparisons were performed by using least significant difference- t test. Results:Compared with the model group, the pomegranate peel polyphenol groups and positive control group showed marked improvement in cysts, desquamation, crusts and epidermal keratinization, and reduced infiltration with inflammatory factors in the dermis at the modeling site. The serum level of IL-17 was significantly lower in the low-, medium- and high-dose pomegranate peel polyphenol groups (61.03 ± 5.99 ng/L, 55.35 ± 2.24 ng/L, 54.35 ± 4.29 ng/L, respectively) , positive control group (48.11 ± 4.07 ng/L) and blank group (42.10 ± 5.62 ng/L) than in the model group (70.24 ± 3.30 ng/L; t = 3.12, 5.34, 5.70, 8.29, 10.54, respectively, all P<0.05) . Immunohistochemical study revealed that the HIF-1α expression level was significantly lower in the low-, medium- and high-dose pomegranate peel polyphenol groups (0.29 ± 0.05, 0.29 ± 0.03, 0.33 ± 0.02, respectively) and positive control group (0.30 ± 0.01) than in the model group (0.41 ± 0.04; t = 4.89, 5.50, 3.62, 5.21, respectively, all P<0.05) ; the RORγt expression level was significantly lower in the low- and high-dose pomegranate peel polyphenol groups (0.28 ± 0.02, 0.31 ± 0.04, respectively) than in the model group (0.35 ± 0.02, t = 3.68, 2.18, respectively, both P<0.05) ; there was no significant difference in the mTOR expression level among these groups ( P = 0.119) . Conclusion:Pomegranate peel polyphenols could improve inflammatory reactions in the rat auriclular model of acne, which may be related to the down-regulation of HIF-1α/RORγt signaling pathway.

Galactinol synthase (GolS) is a key enzyme in the biosynthetic pathway of raffinose family oligosaccharides (RFOs) and plays an important role in plant responses to abiotic stresses. However, the molecular characteristics of the GolS family members in soybean was not well-known. In this study, six members of GmGolS gene family were genome-widely identified, and their physicochemical properties, chromosomal localization, evolutionary relationship, gene structure, conserved motifs, secondary structure, tertiary structure, tissue-specific expression patterns and the expression levels under salt and drought stresses were analyzed. The results showed that six soybean GolS genes were unevenly distributed on four chromosomes, the range of the isoelectric points of six GmGolS proteins was 5.45-6.08, the molecular weight range was 37 567.07-38 817.59 Da, and the number of amino acids was 324-339 aa. The results of subcellular localization showed that 4 proteins were located in the chloroplast, and 2 proteins in the cytoplasm. Phylogenetic tree analysis showed that the members of the soybean GolS gene family were closely adjacent to each other, and were evolutionarily conservative. Six gene members contain 3 or 4 exons. Prediction of secondary and tertiary structures showed that the spatial structure of proteins of all family members was mainly composed of α-helix and random coil structure, with less β-turn and extended chain structure. Tissue-specific expression analysis showed that six GmGolS members expressed to variable degrees in seeds, roots, root hairs, flowers, stems, pods, nodules and leaves. Expression analysis based on qRT-PCR showed that all GmGolS genes showed different degrees of up-regulated expression under salt and drought treatment, indicating that these genes may be related to the response of plants to salt-tolerance and drought-resistance. These results may facilitate subsequent functional analysis of soybean GolS genes.
Droughts ; Soybeans/genetics* ; Phylogeny ; Plant Proteins/metabolism* ; Stress, Physiological/genetics* ; Plants/metabolism* ; Gene Expression Regulation, Plant