Objective To investigate the autophagy effect and the crosstalk with apoptosis by UV in A375 cells.Methods GFP-RFP-LC3 lentivirus were used to evaluate the effect of autophagy after being irradiated with UV of different doses (0、10、30、50 mJ/cm2).After being treated with 30 mJ/cm2 irradiation,the apoptosis rate of A375 with or without autophagy inducer was evaluated by annexin V-FITC/PI with flow cytometry.Western blot was used to distinguish the biomarker of autophagy (BECN,LC3) and apoptosis (Caspase 3,9).Results After being irradiated with 10 mJ/cm2 or 30 rnJ/cm2 UV,the autophagosome was observed at 6 h and rich at 9 h.However,the dots of autophagy had been abundant continually from 3 h with 50 mJ/cm2.The ability of inducing autophagy of UVB is stronger than UVA.UVA and UVB showed synergistic effect in autophagy with the dose of 30 mJ/cm2.The Caspase 3 and Caspase 9 proteins were downregulated after 30 mJ/cm2 UV irradiation with autophagy biomarkers increasing,whereas the apoptosis biomarkers were enriched with the inhibition of autophagy.Conclusions UV can induce autophagy with more significant effect of UVB.Autophagy paly protective role by delaying apoptosis after 30 mJ/cm2 UV irradiation in A375.

Objective To explore the role of 14-3-3σ in cell cycle arrest,proliferation inhibition of HaCaT cells after UVB exposure.Methods Crystal violet assay was used to determine the viable density of HaCaT,HaCaTKD cells after being irradiated with UVB of different doses (10,20,30,40,50,60 and 80 mJ/cm2) for 48 h.After HaCaT and HaCaTKD being treated with 30 mJ/cm2irradiation,cell growth and cell cycle distribution were detected by CCK-8 assay and PI staining combined with flow cytometry,respectively.Western blot was used to evaluate the protein expression of 14-3-3σ,Cdc2,Cdc25c and Cyclin B1.Results After 48 h,the survival rate of both HaCaT and HaCaTKD decreased in a dosedependent manner.Especially,HaCaTKD cells had drastically low proliferation rate compared with normal HaCaT at 10 mJ/cm2 (t =8.83-49.63,P ＜ 0.05).The proliferation rate of HaCaTKD cells was significantly lower than that of HaCaT cells (F =32.89,P ＜ 0.05).After treatment with 30 mJ/cm2 UV irradiation,the ability of proliferation in normal HaCaT cells was recovered after 24 h while there was no proliferation in HaCaTKD cells within 72 h after the same treatment.The distribution of cell cycle has little change in HaCaTKD (P ＞ 0.05).UVB treatment led to cell cycle arrest from 6 to 18 h in HaCaT cells(t =7.41,9.22,9.16,P ＜0.05)while no cell cycle arrest could be observed in the HaCaTKID cell.Western blot detection indicated that the expression of 14-3-3σ in HaCaT cells was upregulated(t =5.42-9.57,P ＜0.05)while the Cdc25c and Cyclin B1 proteins were downregulated in both HaCaT and HaCaTKD cells (t =3.95-11.21,P ＜0.05).Specifically,Cdc2 protein decreased in HaCaT cells(t =4.93-5.37,P ＜ 0.05)but there was no change in HaCaT~ cells.Conclusions 14-3-3σ protein affects the proliferation and cell cycle of HaCaT cells after UVB irradiation.14-3-3oσ co-activates the expression of Cdc2,Cdc25cand Cyclin B1 to mediate UVB-induced G2/M arrest in HaCaT cells.

Objective:To evaluate the relationship between microRNA-93-5p and mitochondrial fusion protein-2 (Mfn2) in mouse nerve cells subjected to oxygen-glucose deprivation and reoxygenation (OGD/R).Methods:Mouse neuroblastoma cells were cultured in vitro to logarithmic growth phase.Experiment Ⅰ Cells were divided into 5 groups ( n=20 each) by the random number table method: control group (group C), group OGD/R, miR-93-5p inhibitor group (group I), siRNA-mfn2 plus miR-93-5p group (group siMfn2+ I) and miR-93-5p negative control group (group NC). Oxygen-glucose deprivation: the cells were cultured for 3 h in a low-glucose balanced salt solution at 37 ℃ in an environment of 5% CO 2-95% N 2.Restoration of oxygen and glucose: the cells were cultured in normal medium at 37℃ in 5% CO 2-95% air for 24 h. Group I, group siMfn2+ I and group NC were transfected with miR-93-5p inhibitor, miR-93-5p inhibitor plus siRNA-mfn2 and negative control miRNA, respectively, at 48 h before the OGD/R model was developed.Cell viability was measured by CCK-8 assay.Cell apoptosis rate was measured by flow cytometry.Quantitative real-time polymerase chain reaction was used to detect the expression of miR-93-5p and Mfn2 mRNA.Western blot was used to detect Mfn2 protein expression.Experiment Ⅱ The wild-type (WT)-Mfn2 and mutant (MUT)-Mfn2 were constructed and transfected into neuroblastoma cells with miR-93-5p mimic and miR-93-5p blank control (miR-93-5pNC), respectively.The cells were divided into 4 groups ( n=5 each) after 48 h of transfection by the random number table method: miR-93-5p NC-WT-Mfn2 co-transfection group, miR-93-5p mimic-WT-Mfn2 co-transfection group, miR-93-5p NC-MUT-Mfn2 co-transfection group, and miR-93-5p mimic-MUT-Mfn2 co-transfection group.The activity of luciferases was measured by double luciferase assay. Results:Experiment Ⅰ Compared with group C, the cell viability was significantly decreased, the apoptosis rate of cells was increased, the expression of miR-93-5p was up-regulated, and the expression of Mfn2 protein and mRNA was down-regulated in the other gorups ( P<0.05). Compared with group OGD/R or group NC, the cell viability was significantly increased, the apoptosis rate of cells was decreased, miR-93-5p expression was down-regulated, and the expression of Mfn2 protein and mRNA was up-regulated in group I ( P<0.05). Compared with group I, the cell viability was significantly decreased, the apoptosis rate of cells was increased, and the expression of Mfn2 protein and mRNA was down-regulated in group siMfn2+ I ( P<0.05). Experiment Ⅱ Compared with miR-93-5p NC-WT-Mfn2 co-transfection group, the luciferase activity was significantly decreased in miR-93-5p mimic-WT-Mfn2 co-transfection group ( P<0.05). There was no significant difference in luciferase activity between miR-93-5p NC-MUT-Mfn2 co-transfection group and miR-93-5p mimic-MUT-Mfn2 co-transfection group ( P>0.05). Conclusions:The miR-93-5p expression is up-regulated, which further targetedly down-regulates the expression of Mfn2, and this may be a mechanism of OGD/R in mouse nerve cells.

Objective:To evaluate the efficacy of holographic image technology in robot-assisted laparoscopic radical prostatectomy (RARP).Methods:The clinical data of 34 patients with prostate cancer who underwent RARP in our hospital during October 2020 and December 2020 was analyzed retrospectively. The average age of the patients was 67.8 (52-78) years. The mean BMI was 25.8 (18.0-32.3) kg/m 2. The median level of PSA before surgery was 13.4 (2-149) ng/ml. Median prostate volume was 31.7 (9.5-159.1) ml. EAU risk groups for biochemical recurrence of localised and locally advanced prostate cancer were list as below: 5 cases of low-risk, 7 cases of medium-risk, 22 cases of high-risk. There were 9, 16, 9 cases with the ASA score of 1, 2, 3 point, respectively. Preoperative Gleason score of 34 patients were list as below: 9 cases in score ≤6 group, 15 cases in score=7 group, 10 cases in score ≥8 group. For clinical stage before the surgery, 13 cases ≤T 2a stage, 1 case in T 2b stage, 20 cases ≥T 2c stage. The engineers established holographic images of 34 patients based on multiparametric magnetic resonance imaging (mpMRI) and the reports before the operation. Surgeons can obtain the size and location of tumors, surrounding neurovascular bundles visually by revolving, assembling, disassembling and concealing images, which was helpful for pre-surgery planning. By manipulating the holographic images extracorporeally, surgeons can discriminate Internal sphincter of urinary bladder and vesicoprostatic muscle, neurovascular bundles, membranous part, seminal vesicle easily, which improves the operation accuracy. Results:All 34 cases underwent operation successfully without transferring to open surgery. The median operative time was 157.5 (95-276) min with an estimated blood loss of 50 (20-300) ml. The median drainage removal time was 2 d and median hospitalization time was 3.5 d, respectively. The catheters were removed within an average time of 20.5 d. For postoperative Gleason score, there were 2 cases in score ≤6 group, 16 cases in score =7 group, 8 cases in score ≥8 group and 8 cases can’t make a score. For clinical stage after the surgery, 10 cases were ≤T 2a stage, 1 case was T 2b stage, 23 cases were ≥cT 2c stage. 22 cases underwent pelvic lymph node dissection, including a patient with right iliac fossa lymph node metastasis. There were 2 cases with positive surgical margin and 3 cases with Clavien-DindoⅠcomplications. The rate of 1-month and 3-month urinary continence were 47.1% and79.4%, respectively, 8 cases recovered erectile function after 3 month. Conclusions:Holographic image technology can promote cancer dissection completely, achieve urinary continence early and reduce perioperative complications tremendously. The technology is the "intraoperative security" for the accurate surgical treatment of prostate cancer.

OBJECTIVETo observe the impacts of electroacupuncture (EA) at Zusanli (ST 36) and Feishu (BL 13) applied 30 min before the operation till the end of the operation on the postoperative inflammatory reaction and pulmonary complications in the senile patients after radical resection of pulmonary carcinoma.

METHODSEighty senile patients of pulmonary carcinoma were selected and randomized into an observation group and a control group, 40 cases in each one. In the observation group, EA stimulation at Zusanli (ST 36) and Feishu (BL 13) was used 30 min before the operation till the end of the operation. In the control group, electric stimulation was not used. Separately, before operation (T, basic state), 12 h after operation (T) and 24 h after operation (T), blood sample was collected from the central vein. The concentrations of plasma tumor necrosis factor-ɑ (TNF-ɑ) and interleukin-10 (IL-10) were detected. Additionally, the radial arterial blood sample was collected at the above time points; oxygen partial pressure (PaO) was determined; pulmonary alveoli-arterial partial pressure of oxygen (PDO) and oxygenation index (OI) were calculated. The pulmonary complication in the two days after operation was recorded.

RESULTSCompared with the control group, in the observation group, at Tand T, TNF-ɑ concentration and PDOwere lower (all<0.05); plasma IL-10 concentration and OI were higher (all<0.05). In the observation group, the incidences of postoperative pneumonia and acute pulmonary injury were lower than those in the control group (both<0.05).

CONCLUSIONSEA reduces the postoperative inflammatory reaction in the senile patients with radical resection of pulmonary carcinoma and decreases the postoperative pulmonary complicattizen.

【Objective】 To investigate the correlation of monocytes and high-density lipoprotein cholesterol ratio(MHR) and albumin with the severity of coronary artery lesions in patients with unstable angina pectoris. 【Methods】 We enrolled 342 patients with unstable angina pectoris. According to the Gensini score of their coronary angiography results, they were divided into Gensini≤ 20 group, 2040 group. The differences in biochemical indicators between the groups were compared, and the correlation between the different indicators and the Gensini score was analyzed. According to the MHR quartile grouping, there were differences between the comparison groups. LDL-C was divided into subgroups and then subjected to multifactor Logistic regression analysis. 【Results】 MHR differed significantly among low, moderate and high grade lesions (P<0.05). Subgroup analysis showed that in low LDL-C group, Gensini score was positively correlated with MHR(P<0.05), while in high LDL-C group, Gensini score was negatively correlated with albumin(P<0.05). Multivariate Logistic regression analysis showed that the MHR level of patients with high Gensini score was 102.375 times higher than that of patients with low Gensini score(P<0.05). In the group with high LDL-C, the serum albumin level in the group with low Gensini score was 1.431 times that in the group with high Gensini score and 1.218 times that in the group with moderate Gensini score(all P<0.05). 【Conclusion】 In patients with unstable angina pectoris, especially when LDL-C levels are not high, both high MHR and low serum albumin are independent risk factors for the severity of coronary artery disease.

【Objective】 Corin, a transmembrane serine protease that can cleave atrial natriuretic peptide precursor (pro-ANP) into atrial natriuretic peptide with smaller bioactive molecules, participates in the pathophysiological process of hypertension and cardiac hypertrophy. The purpose of this study was to explore the relationship of Corin gene variation with blood pressure responses to sodium and potassium dietary interventions. 【Methods】 In 2004, we recruited 514 participants from 124 families in 7 villages of Baoji, Shaanxi Province, China. All the subjects received a 3-day normal diet, a 7-day low-salt diet, a 7-day high-salt diet, and finally a 7-day high-salt and potassium supplementation. Fifteen single nucleotide polymorphisms (SNPs) of Corin gene were selected for final analysis. 【Results】 SNPs rs12509275 were significantly associated with diastolic blood pressure (DBP) response to low-salt diet, while rs3749584 was associated with pulse pressure (PP) response to low-salt diet.SNP rs3749584 and rs10517195 were significantly associated with PP response to high-salt diet. In addition,rs17654278 were significantly associated with systolic blood pressure (SBP) response to high-salt and potassium supplementation, rs2271037 was significantly correlated with DBP responses to high-salt and potassium supplementation, and rs4695253, rs12509275, rs2351783, rs36090894 were significantly associated with PP response to high-salt and potassium supplementation. 【Conclusion】 Corin gene polymorphisms were associated with blood pressure response to sodium and potassium, suggesting that Corin gene may be involved in pathophysiological process of salt sensitivity and potassium sensitivity.

【Objective】 Dyslipidemia has shown to be associated with cardiovascular, metabolic and renal diseases. This study aimed to investigate the association between residual cholesterol and the risk of subclinical renal damage (SRD). 【Methods】 A total of 2 342 participants were recruited from the previously established Hanzhong Adolescent Hypertension Study cohort. According to estimated glomerular filtration rate(eGFR) and urinary albumin-to-creatine ratio(uACR), the subjects were divided into SRD group and non-SRD group. The associations of residual cholesterol with eGFR, uACR, and the risk of SRD were analyzed by multiple linear and Logistic regression analyses. 【Results】 Residual cholesterol was positively correlated with uACR(r=0.081, P<0.001) but negatively correlated with eGFR (r=-0.091, P<0.001). Multiple linear regression analysis revealed that residual cholesterol was an influencing factor of uACR (β=0.075, P<0.001) and eGFR (β=-0.027, P<0.001) after adjustment for gender, age, smoke, alcohol, exercise, BMI, hypertension, diabetes and serum uric acid. In addition, Logistic regression analysis revealed that residual cholesterol was significantly associated with the risk of SRD independently of potential confounders [OR(95% CI)=1.387 (1.113-1.728), P<0.001]. Further subgroup analysis showed that residual cholesterol was significantly associated with the risk of SRD in women but not in men. 【Conclusion】 Residual cholesterol is a contributing factor in the risk of subclinical renal damage with gender-specific association.

【Objective】 4-like protein with down-regulated expression and development in neural precursor cells (NEDD4L) plays an important role in blood pressure (BP) regulation and sodium homeostasis by regulating epithelial sodium channel protein. In this study, we aimed to explore the relationship of NEDD4L gene polymorphisms with BP responses to sodium and potassium intake. 【Methods】 In 2004, 514 subjects from 124 families in Meixian County, Shaanxi Province, were recruited to establish a salt-sensitive hypertension study cohort. All the subjects received a 3-day baseline survey, a 7-day low-salt diet, a 7-day high-salt diet, and finally a 7-day high-salt and potassium supplementation. Their BP was measured and peripheral blood samples were collected at different intervention periods. The 14 gene polymorphisms of NEDD4L gene were genotyped and analyzed by MassARRAY platform. 【Results】 BP decreased on a low-salt diet, and significantly increased on a high-salt diet, and decreased again after potassium supplementation. NEDD4L SNPs rs74408486 were significantly associated with systolic BP, diastolic BP and mean arterial pressure responses to the low-salt diet. SNPs rs292449 and rs2288775 were significantly associated with pulse pressure response to the high-salt diet. In addition, SNPs rs563283 and rs292449 were significantly associated with diastolic BP, mean arterial pressure, and pulse pressure responses to high-salt and potassium supplementation diet. 【Conclusion】 NEDD4L gene polymorphisms were significantly associated with BP responses to sodium and potassium intake, suggesting that NEDD4L gene may be involved in the development of salt sensitivity and potassium sensitivity.

【Objective】 Based on our previously established salt-sensitive hypertension cohort, we aimed to examine the association of genetic variants in uromodulin with blood pressure(BP) responses to dietary interventions of sodium and potassium intake. 【Methods】 In 2004, 514 subjects from 124 families in Mei County, Shaanxi Province, were recruited to establish the salt-sensitive hypertension study cohort. Among them, 333 non-parent subjects were selected and sequentially maintained on a normal-diet for 3 days, low-salt diet for 7 days, then a high-salt diet for 7 days and a high-salt diet with potassium supplementation for another 7 days. Thirteen single nucleotide polymorphisms(SNPs) in the uromodulin gene were genotyped on the MassARRAY platform. 【Results】 BP levels decreased from the baseline to low-salt diet, increased from low-salt to high-salt diet, and decreased again from the high-salt diet to the high-salt plus potassium supplementation intervention. SNPs rs77875418 and rs4997081 of the uromodulin gene were significantly associated with diastolic BP(DBP) and mean arterial pressure(MAP) responses to high-salt diet. In addition, SNPs rs77875418, rs79245268, rs4293393, rs6497476, rs4997081, rs13333226, and rs12917707 were significantly associated with systolic BP(SBP), DBP, and MAP responses to high-salt diet with potassium supplementation. 【Conclusion】 Genetic variants in uromodulin gene are significantly associated with BP responses to sodium and potassium supplementation, suggesting that uromodulin may be mechanistically involved in BP sodium-sensitivity and potassium-sensitivity.