Objective:To explore the possible effects of Bushen Huoxue Formula (the kidney tonifying and blood activating prescription) on the proliferation and extracellular matrix synthesis of nucleus pulposus cells by regulating the circ_0036763/miR-583 axis.Methods:Real time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression levels of circ0036763 and miR-583 in normal and intervertebral disc degeneration (IDD) nucleus pulposus cells; IDD nucleus pulposus cells were divided into pcDNA group, pcDNA circ_0036763 group, pcDNA circ_0036763+ mimic NC group, and pcDNA circ_0036763+ miR-583 mimic group. qRT-PCR was used to detect the expression levels of circ_0036763 and miR-583 in nucleus pulposus cells in each group, methyl thiazolyl tetrazolium (MTT) was used to detect cell proliferation (A value), and Western blot was used to detect the expression of proliferating cell nuclear antigen (PCNA), collagen Ⅰ, and collagen Ⅱ proteins in nucleus pulposus cells, The dual luciferase assay reported experimental validation of the targeting relationship between circ_0036763 and miR-583. 27 mice were divided into sham surgery group, IDD group, and kidney tonifying and blood activating formula group. IDD models were established in all groups except for the sham surgery group. After successful modeling, the sham surgery group and IDD group were given physiological saline by gavage, while the kidney tonifying and blood activating formula group was given 1.5 g/ml of kidney tonifying and blood activating formula by gavage for 3 consecutive weeks. QRT-PCR was used to detect the expression levels of circ0036763 and miR-583 in the nucleus pulposus cells of mice in each group, MTT was used to detect cell proliferation, and Western blot was used to detect the expression of PCNA, collagen Ⅰ, and collagen Ⅱ proteins.Results:The expression level of circ_0036763 in IDD nucleus pulposus cells decreased, while the expression level of miR-583 increased (all P<0.05); Overexpression of circ_0036763 can promote proliferation and extracellular matrix synthesis of nucleus pulposus cells (all P<0.05); Circ_0036763 targets miR-583 and upregulates miR-583 reversible overexpression. Circ_0036763 enhances the proliferation and extracellular matrix synthesis ability of IDD nucleus pulposus cells. Compared with the sham surgery group, the IDD group showed an increase in collagen Ⅰ protein expression and miR-583 expression levels (all P<0.05), while the cell A value, PCNA and collagen Ⅱ protein expression, and circ_0036763 expression levels decreased (all P<0.05); Compared with the IDD group, the Kidney Tonifying and Blood Activating Formula group showed a decrease in collagen Ⅰ protein expression and miR-583 expression levels (all P<0.05), while the cell A value, PCNA and collagen Ⅱ protein expression, and circ_0036763 expression levels increased (all P<0.05). Conclusions:The kidney tonifying and blood activating formula (Bushen Huoxue) may induce proliferation and extracellular matrix synthesis of nucleus pulposus cells by regulating the circ_0036763/miR-583 axis.

Objective: To find out the differences in gene characteristics between cervical cancer patients with and without lymph node metastasis, and to provide reference for therapy. Methods: From January 2018 to June 2022, recurrent cervical cancer patients 39 cases with lymph node metastasis and 73 cases without lymph node metastasis underwent testing of 1,021 cancer-related genes by next-generation sequencing. Maftools software was used to analyze somatic single nucleotide/insertion-deletion variation mutation, co-occurring mutation, cosmic mutation characteristics, oncogenic signaling pathways. Results: EP300 and FBXW7 were significantly enriched in lymph node-positive patients.Lymph node-positive patients with EP300 or FBXW7 mutations had lower overall survival (OS) after recurrence. Both lymph node-positive and -negative patients had plenty of co-occurring mutations but few mutually exclusive mutations. Lymph node-positive cooccurring mutation number ≥6 had lower OS, while lymph node-negative co-occurring mutation number ≥3 had lower OS after recurrence. The etiology of SBS3 was defects in DNA double strand break repair by homologous recombination, which exclusively exist in lymph node-positive patients. There was no difference in median tumor mutation burden (TMB) between positive and negative lymph nodes, but TMB was significantly associated with PIK3CA mutation. Conclusion The somatic SNV/Indels of EP300 and FBXW7, SBS3 homologous recombination-mediated DNA repair defect were enriched in lymph node-positive patients.For lymph node-positive patients, EP300 or FBXW7 mutations predicted poor prognosis. No matter lymph node-positive or negative, more co-occurring mutation number predicted poor prognosis. PIK3CA mutation may account for the higher TMB and help identify patients who benefit from immunotherapy.

Objective: To find out the differences in gene characteristics between cervical cancer patients with and without lymph node metastasis, and to provide reference for therapy. Methods: From January 2018 to June 2022, recurrent cervical cancer patients 39 cases with lymph node metastasis and 73 cases without lymph node metastasis underwent testing of 1,021 cancer-related genes by next-generation sequencing. Maftools software was used to analyze somatic single nucleotide/insertion-deletion variation mutation, co-occurring mutation, cosmic mutation characteristics, oncogenic signaling pathways. Results: EP300 and FBXW7 were significantly enriched in lymph node-positive patients.Lymph node-positive patients with EP300 or FBXW7 mutations had lower overall survival (OS) after recurrence. Both lymph node-positive and -negative patients had plenty of co-occurring mutations but few mutually exclusive mutations. Lymph node-positive cooccurring mutation number ≥6 had lower OS, while lymph node-negative co-occurring mutation number ≥3 had lower OS after recurrence. The etiology of SBS3 was defects in DNA double strand break repair by homologous recombination, which exclusively exist in lymph node-positive patients. There was no difference in median tumor mutation burden (TMB) between positive and negative lymph nodes, but TMB was significantly associated with PIK3CA mutation. Conclusion The somatic SNV/Indels of EP300 and FBXW7, SBS3 homologous recombination-mediated DNA repair defect were enriched in lymph node-positive patients.For lymph node-positive patients, EP300 or FBXW7 mutations predicted poor prognosis. No matter lymph node-positive or negative, more co-occurring mutation number predicted poor prognosis. PIK3CA mutation may account for the higher TMB and help identify patients who benefit from immunotherapy.

Objective: To find out the differences in gene characteristics between cervical cancer patients with and without lymph node metastasis, and to provide reference for therapy. Methods: From January 2018 to June 2022, recurrent cervical cancer patients 39 cases with lymph node metastasis and 73 cases without lymph node metastasis underwent testing of 1,021 cancer-related genes by next-generation sequencing. Maftools software was used to analyze somatic single nucleotide/insertion-deletion variation mutation, co-occurring mutation, cosmic mutation characteristics, oncogenic signaling pathways. Results: EP300 and FBXW7 were significantly enriched in lymph node-positive patients.Lymph node-positive patients with EP300 or FBXW7 mutations had lower overall survival (OS) after recurrence. Both lymph node-positive and -negative patients had plenty of co-occurring mutations but few mutually exclusive mutations. Lymph node-positive cooccurring mutation number ≥6 had lower OS, while lymph node-negative co-occurring mutation number ≥3 had lower OS after recurrence. The etiology of SBS3 was defects in DNA double strand break repair by homologous recombination, which exclusively exist in lymph node-positive patients. There was no difference in median tumor mutation burden (TMB) between positive and negative lymph nodes, but TMB was significantly associated with PIK3CA mutation. Conclusion The somatic SNV/Indels of EP300 and FBXW7, SBS3 homologous recombination-mediated DNA repair defect were enriched in lymph node-positive patients.For lymph node-positive patients, EP300 or FBXW7 mutations predicted poor prognosis. No matter lymph node-positive or negative, more co-occurring mutation number predicted poor prognosis. PIK3CA mutation may account for the higher TMB and help identify patients who benefit from immunotherapy.

Objective:To investigate the risk factors for concomitant cardiac autonomic neuropathy in diabetic patients and to develop a Nomogram prediction model.Methods:One hundred and fifty-eight diabetic patients admitted to in Southern Hospital Zengcheng Branch from March 2019 to March 2021 were selected. Patients with normal heart rate variability were the diabetic group, and patients with abnormal heart rate variability were the group with diabetes mellitus complicated by cardiac autonomic neuropathy. Logistic regression analysis was used to analyze the risk factors of cardiac autonomic neuropathy. Nomogram models were developed and model performance was evaluated. Decision curve analysis (DCA) was used to assess the net clinical benefit of the Nomogram model.Results:Comparison of general data showed that fasting blood glucose, tumour necrosis factor-α (TNF-α), glomerular filtration rate (eGER), uric acid, C-reactive protein (CRP), interleukin-6 (IL-6), free fatty acids (FFA), standard deviation of sinus heart beat RR interval (SDNN), and duration of diabetes compared to the diabetic group had statistically significant ( P<0.05); the results of the subject work characteristics (ROC) curve analysis showed that the best cut-off values for fasting glucose, TNF-α, eGFR, uric acid, CRP, IL-6, FFA, SDNN and duration of diabetes were >7.53 mmol/L, >98.45 ng/L, ≤94.79 ml/(min·1.73 m 2), > 87.3 μmol/L, >6.22 μmol/L, >37.84 ng/L, >839.19 μmol/L, ≤ 95.88 ms, >9 years; multi-factorial Logistic regression analysis showed that fasting glucose (>7.53 mmol/L), TNF-α (>98.45 ng/L), CRP (>6.22 μmol/L), IL-6 (>37.84 ng/L), FFA (>839.19 μmol/L), SDNN (≤95.88 ms), and duration of diabetes (>9 years) were risk factors for the development of cardiac autonomic neuropathy in diabetic patients; internal validation showed that the Nomogram model predicted a C-index of 0.706 (95% CI 0.668 - 0.751) for the risk of cardiac autonomic neuropathy. The DCA results showed that the Nomogram model predicted a risk threshold of >0.25 for the development of cardiac autonomic neuropathy and that the Nomogram model provided a net clinical benefit. Conclusions:There are many risk factors for cardiac autonomic neuropathy, and the nomogram model based on risk factors in this study has good predictive power and may provide a reference for clinical screening of high-risk patients and further improvement of treatment planning.

Objective: To understand the status of teaching environmental health in primary and secondary schools and explore its association with myopia, so as to provide a basis for precision myopia prevention and control. Methods: From September to October 2020, a stratified cluster sampling method was used in 16 districts of Shanghai. After matching the teaching environment testing classrooms with the classrooms where the students were surveyed, totally 470 classrooms and 14 624 students were monitored. Trained professional technicians used laser range finder, luxmeter to measure parameters about teaching environmental health. Students were subject vision testing and questionnaires. Multilevel models was used to analyze the association between teaching environment and screening myopia. Results: The qualified rates of each indicators of teaching environmental health were as follows: window to floor area ratio (93.0%), classroom area per capita (82.1%), uniformity ratio of illuminance of blackboard ( 79.7% ), blackboard size (68.7%), daylight factor (67.0%), blackboard hanging height (66.9%), average illuminance on desk (66.4%), uniformity ratio of illuminance of desk (63.6%), reflectance of blackboard (46.8%), average illuminance on blackboard (41.4%), reflectance of wall (33.2%), desk and chair allocation compliance rate (19.8%). The results of two level Logistic model analysis showed that unqualified average illuminance on blackboard was positively associated with screening myopia ( OR =1.24, 95% CI = 1.05 -1.47, P <0.05). Conclusion Teaching environmental health of primary and secondary schools in Shanghai is not optimistic, teaching environment especially the desk and chair allocation compliance rate need to be improved. The average illumination on blackboard is positively correlated with screening myopia, suggesting that myopia prevention and control can focus on improving the illuminance on blackboard.

Background: Cervical cancer is still present a major public health problem, especially in developing countries. In International Federation of Gynaecology and Obstetrics 2018, allowing assessment of retroperitoneal lymph nodes by imaging and/or pathological findings and, if deemed metastatic, the case is designated as stage IIIC (with r and p notations). Patients with lymph node metastases have lower overall survival (OS), progression free survival (PFS), and survival after recurrence, especially those who have unresectable macroscopical positive lymph nodes. Retrospective analysis suggests that there may be a benefit to debulking macroscopic nodes that would be otherwise difficult to sterilize with standard doses of radiation therapy. However, there are no prospective study reporting that resecting macroscopic nodes before concurrent chemoradiation therapy (CCRT) would improve PFS or OS of cervical cancer and no guidelines for surgical resection of bulky lymph nodes. The CQGOG0103 study is a prospective, multicenter and randomized controlled trial (RCT) evaluating lymph node dissection on stage IIICr of cervical cancer. Methods Eligible patients are histologically confirmed cervical squamous cell carcinoma, adenocarcinoma, adeno-squamous cell carcinoma. Stage IIICr (confirmed by computed tomography [CT]/magnetic resonance imaging/positron emission tomography/CT) and the short diameter of image-positive lymph node ≥15 mm. 452 patients will be equally randomized to receive either CCRT (pelvic external-beam radiotherapy [EBRT]/extended-field EBRT + cisplatin [40 mg/m2] or carboplatin [the area under curve=2] every week for 5 cycles + brachytherapy) or open/minimally invasive pelvic and para-aortic lymph node dissection followed by CCRT. Randomization is stratified by status of para-aortic lymph node. The primary endpoint is PFS. Secondary endpoints are OS and surgical complications. A total of 452 patients will be enrolled from multiple hospitals in China within 4 years and followed up for 5 years.

Objective:To investigate the effects of excessive fluoride exposure on astrocytes and the expression of glial fibrillary acidic protein (GFAP), in vitro and in vivo. Methods:(1) In vivo experiment: 24 SPF SD rats, half male and half female, were randomly divided into control and fluoride exposed groups according to sex and body weight, 12 rats in each group. Rats were fed with < 1 mg/L and 50 mg/L sodium fluoride solution prepared by tap water for 6 months, respectively. The expression levels of GFAP protein in rat brain tissue were measured by immunofluorescence, immunohistochemistry and Western blotting. (2) In vitro experiment: adult (6-month-old) rat cortical astrocytes were extracted and cultured in primary culture (4 mmol/L sodium fluoride solution for 24 h), and the astrocytes were identified by immunofluorescence, and GFAP mRNA and protein expression levels were detected by real-time fluorescence quantitative PCR and Western blotting, respectively, and astrocytes apoptosis and calcium ion content were detected by flow cytometry. Results:(1) In vivo experiment: the results of immunofluorescence, immunohistochemistry and Western blotting showed that the GFAP protein expression level in brain tissue of rats exposed to fluoride was higher than that of control group (0.440 ± 0.200 vs 0.250 ± 0.120, t =-5.93, P = 0.027; 0.270 ± 0.020 vs 0.240 ± 0.050, t =-4.87, P = 0.040; 1.017 ± 0.001 vs 0.486 ± 0.006, t =-52.48, P = 0.001). (2) In vitro experiment: GFAP positive cells were identified as astrocytes by immunofluorescence; GFAP mRNA expression level was higher in fluoride exposed group than that of control group by real-time fluorescence quantitative PCR (2.780 ± 0.120 vs 0.134 ± 0.005, t =-37.84, P = 0.001). The Western blotting results showed that the GFAP protein expression level was higher in fluoride exposed group than that of control group (2.76 ± 0.10 vs 1.38 ± 0.05, t =-20.44, P = 0.002). Flow cytometry results showed that the apoptosis rate of astrocytes was higher in fluoride exposed group than that of control group (%: 55.0 ± 1.0 vs 3.5 ± 0.6, t =-10.28, P = 0.009) and the calcium ion content was lower than that of control group (%: 54 ± 9 vs 72 ± 13, t = 4.64, P = 0.043). Conclusion:Excessive fluoride exposure causes increased GFAP expression in astrocytes in vitro and in vivo, promotes apoptosis, and affects calcium signaling pathways.

Objectives:To investigate the epidemiological features and associated factors of chronic renal insufficiency (CRI) in Binhai county from Jiangsu province.Methods:This is a cross-sectional study including individuals aged≥18 years old and participating in health examinations of Binhai county from January to December 2018. Medical records were collected to analyze the epidemiology of CRI [estimated glomerular filtration rate <60 ml·min -1·(1.73 m 2) -1]. Multivariate logistic regression was used to analyze the associated influencing factors of CRI. Results:A total of 395 541 individuals residing in Binhai county were enrolled, with 190 258 males (48.1%) and age of (55.34±15.12) years old. The overall crude prevalence of CRI was 2.04% (8 065/395 541, 95% CI 2.00%-2.08%) in this adult population. Furthermore, the age- and gender-standardized overall prevalence of CRI was 1.22% (95% CI 1.18%-1.25%), with a rate of 1.47% (4 676/205 283, 95% CI 1.42%-1.52%) in women and a rate 0.95% (3 389/190 258, 95% CI 0.91%-1.00%) in men. There was a strong positive correlation between the risk of CRI and age (per 10-year increase, OR=2.449, 95% CI 2.402-2.497). Compared with individuals <30 years old, the OR of CRI in individuals aged 60-69, 70-79 and ≥80 years old were 3.827 (95% CI 3.010-4.864), 12.004 (95% CI 9.457-15.239) and 44.636 (95% CI 35.187-56.622) respectively. Females ( OR=1.142, 95% CI 1.083-1.203), increasing systolic blood pressure (per 10 mmHg increase, OR=1.062, 95% CI 1.048-1.076), increasing heart rate (per 10-beat/min increase, OR=1.071, 95% CI 1.044-1.098), elevating triglyceride (per 1.33 mmol/L increase, OR=1.140, 95% CI 1.119-1.162), elevating fasting blood glucose (5.6-6.9 mmol/L/<5.6 mmol/L, OR=1.158, 95% CI 1.086-1.233; ≥7 mmol/L/<5.6 mmol/L, OR=1.387, 95% CI 1.296-1.484) and central obesity ( OR=1.126, 95% CI 1.068-1.187) were independent risk factors for CRI. Conclusions:The age- and gender-adjusted prevalence of CRI in adults in Binhai county is 1.22%. Older age, females, central obesity, and high levels of triglyceride, systolic blood pressure, heart rate and fasting glucose are independent associated factors of CRI.

Objective: To explore the relationship between myopia and nutritional status among upper elementary students in Shanghai. Methods: A stratified cluster sampling method was used to select 5 576 fourth and fifth grade students from 16 districts in Shanghai, vision test, physical examination and questionnaire survey were administered. Two level multivariate Logistic regression and mixed linear models were used to analyze the relationship between myopia, spherical equivalent (SE) and nutritional status. Results: The detection rates of overweight and obesity were 17.0%(949) and 18.7%(1 042), respectively. There was a statistical difference in the proportion of the nutritional status in boys and girls( χ 2=210.94, P <0.01), with overweight and obesity in boys higher than those of girls. The overweight boys were more likely to have myopia (univariate analysis: OR=1.23, P =0.04; multivariate analysis: OR=1.21, P =0.06) than the normal weight boy. Overweight students tended to had more negative refractive than those with normal weight in both boys and girls(multivariate analysis, boys: β =-0.19, girls: β=-0.24, P <0.05). Myopia risk and refractive power in obese students showed no statistical difference with normal weight students( P >0.05). Conclusion Overweight students have lower SE than the normal weight among upper elementary students of Shanghai, more researches are needed to confirm this finding and to provide theoretical bases for myopia prevention and control.