Objective: To analyze the trends in incidence and mortality of prostate cancer in Ningbo City, Zhejiang Province, so as to provide insights into the prevention and control of prostate cancer. Methods: Data of the incidence and mortality of prostate cancer in Ningbo City from 2011 to 2023 were collected through Ningbo Chronic Disease Collaborative Management System. The incidence and mortality of prostate cancer were calculated and standardized by the data from the sixth national population census in 2010 and the Segi's world standard population in 1960. The trends in incidence and mortality of prostate cancer were evaluated using average annual percent change (AAPC). Results: A total of 15 411 cases of prostate cancer were reported in Ningbo City from 2011 to 2023, and the crude incidence, Chinese-standardized incidence and world-standardized incidence were 39.62/105, 22.18/105 and 16.49/105, respectively, showing upward trends (AAPC=14.782%, 10.390% and 10.608%, all P<0.05). The Chinese-standardized incidence of prostate cancer was higher in urban areas than in rural areas, and both showed upward trends (25.14/105 vs. 19.44/105; AAPC=9.057% and 14.272%, both P<0.05). The crude incidence of prostate cancer in the groups aged 50-<60 years, 60-<70 years, 70-<80 years and ≥80 years showed upward trends (AAPC=11.657%, 14.031%, 10.734% and 5.300%, all P<0.05). A total of 3 739 deaths were reported, and the crude mortality, Chinese-standardized mortality and world-standardized mortality were 9.66/105, 5.23/105 and 3.71/105, respectively, showing upward trends (AAPC=8.458%, 3.620% and 3.602%, all P<0.05). The Chinese-standardized mortality of prostate cancer was higher in urban areas than in rural areas, and both showed upward trends (5.35/105 vs. 5.13/105; AAPC=3.183% and 3.962%, both P<0.05). The crude mortality of prostate cancer the groups ageds ≥80 years showed an upward trend (AAPC=7.482%, P<0.05). Conclusions From 2011 to 2023, the incidence and mortality of prostate cancer in Ningbo City showed upward trends. Special attention should be paid to urban residents, and prostate cancer screening should be strengthened among males aged 50 years and older.

Objective To investigate the effect of afferent blockade of visceral adipose tissue(VAT)on cardiac function and cardiac neural remodeling in rats after myocardial infarction(MI).Methods After 30 healthy SPF-grade male SD rats were subjected,12 of them were randomly divided into control group(n=6)and activation group(n=6).In the activation group,low-dose capsaicin(1 mmol/L)was used to activate VAT afferent nerves,while in the control group,an equal amount of normal saline was injected,and real-time blood pressure and heart rate were monitored for 30 min.The other 18 rats were randomly assigned into sham group(n=6),MI group(n=6),and high-dose capsaicin blockade group(n=6).The MI model was established by ligating the left anterior descending coronary artery.After MI modeling,the high-dose capsaicin blockade group was give 33 mmol/L capsaicin to block VAT afferent nerve,and the sham opera-tion group and MI group were injected with the same amount of normal saline.After 2 weeks,car-diac function was measured by echocardiography,infarct size was measured by TTC staining,heart rate variability was analyzed,and myocardial tyrosine hydroxylase(TH)was measured.The levels of myocardial superoxide dismutase(SOD)and malondialdehyde(MDA)were measured by biochemical methods.Results More significant changes in blood pressure and heart rate were observed in the activation group than the control group(P＜0.01).The MI group had obviously larger infarct size,higher LVEDD and LVESD,and increased myocardial TH density and MDA level,but lower LVEF and myocardial SOD activity than the sham group(P＜0.05).However,the infarct size,LVEDD(9.15±0.37 mm vs 10.1±0.85 mm),LVESD(6.33±0.40 mm vs 7.87±0.86 mm)were obviously decreased,while LVEF[(67.04±3.34)％vs(47.10±3.89)％]and myocar-dial FS[(33.26±2.50)％vs(20.81±2.14)％]activity were greatly increased in the high dose capsaicin group than the MI group(P＜0.05).Conclusion Activation of VAT afferent nerve can increase blood pressure and heart rate;while its blockade can reduce the infarct size,protect cardiac function and inhibit cardiac nerve remodeling in MI rats,possibly by reducing oxidative stress.

Objective:To investigate the effect of tocilizumab (TCZ) on ventricular arrhythmias (VAs) after myocardial infarction (MI) in Sprague-Dawley rats and explore its potential mechanism.Methods:The random number table method was used to divide 32 adult male Sprague-Dawley rats into 4 groups: Sham group, TCZ group, MI group and MI+TCZ group, with 8 rats in each group. The MI model was established by ligation of the left anterior descending branch of the coronary artery in the MI and MI+TCZ groups, and only sutured without ligation in the Sham and TCZ groups. TCZ was injected into the left superior cervical ganglion (SCG) of rats in the TCZ and MI+TCZ groups after successful modeling or sham operation, and the same amount of normal saline was injected in the Sham and MI groups. 24 h after successful modeling, ECG of rats in each group was recorded, heart rate variability (HRV, including low frequency power (LF), high frequency power (HF), LF/HF ratio), QT interval, QTc interval were calculated, and left ventricular effective refractory period (ERP) and VA inducibility were measured. Myocardial infarct size and tissue changes were observed with triphenyl tetrazolium chloride staining and HE staining. Real-time PCR analysis was used to detect the messager RNA (mRNA) expression of interleukin-6 (IL-6) and signal transducer and activator of transcription (STAT) 3 in SCG and potassium voltage-gated channel subfamily D member 2 (Kcnd2) in myocardial infarction periphery. The expression of c-fos in SCG was detected by immunofluorescence staining.Results:Compared with Sham group and MI+TCZ group, rats in MI group had higher LF and LF/HF ratio, longer QT interval and QTc interval, more VAs induced, lower HF and shorter ERP ( P all<0.05). Triphenyl tetrazolium chloride staining and HE staining showed that rats in the Sham and TCZ groups had normal myocardial tissue structure, those in the MI group had severe myocardial injury, and those in the MI+TCZ group had less myocardial injury than those in the MI group. Real-ime PCR analysis showed that compared with Sham group and MI+TCZ group, mRNA expression levels of IL-6 and STAT3 in SCG of rats in MI group were higher, and mRNA expression level of myocardial Kcnd2 was lower ( P all<0.05). Immunofluorescence staining showed that the content of c-fos in SCG of rats in MI group was higher than that of Sham group and MI+TCZ group ( P all<0.05). Conclusions:TCZ may reduce neural activity of the SCG after MI by inhibiting the IL-6/STAT3 signaling pathway, thereby alleviating myocardial injury and inhibiting VAs.

Objective To investigate the correlations of levels of 25-hydroxyvitamin D[25(OH) D], human β-defencin-2 (HBD-2) and insulin-like growth factor 1(IGF-1) in peripheral blood with disease severity of acne patients. Methods A total of 128 patients with acne vulgaris were included in the acne group and 128 healthy subjects were included in the control group. The levels of 25(OH) D, HBD-2 and IGF-1 in peripheral blood of two groups were detected. The relationships between the levels of 25(OH) D, HBD-2, IGF-1 in peripheral blood and the disease severity of acne patients were analyzed. The relationships between 25(OH) D, HBD-2 and IGF-1 levels in peripheral blood of acne patients were analyzed. The diagnostic value of 25(OH) D, HBD-2 and IGF-1 levels in severe acne was evaluated using receiver operating characteristic (ROC) curves. Results The level of 25(OH) D in peripheral blood of the acne group was significantly lower than that of the control group, and the level of IGF-1 was significantly higher than that of the control group (P < 0.05). There was no significant difference in HBD-2 levels between the two groups (P>0.05). In the acne group, with the aggravation of disease severity, the level of 25(OH) D in peripheral blood was significantly decreased, and the level of IGF-1 was significantly increased (P < 0.05). The level of 25(OH) D in peripheral blood of acne patients was negatively correlated with the level of IGF-1 and the severity of the disease (r=-0.509, -0.586, P < 0.05), and the level of IGF-1 was positively correlated with the severity of the disease (r=0.547, P < 0.05). HBD-2 level was not correlated with 25(OH) D, IGF-1 level and disease severity (P>0.05). The sensitivity of peripheral blood 25(OH) D level and IGF-1 level in evaluating severity acne was 94.12%, 61.26%, respectively, and the specificity was 70.59%, 77.48%, respectively. The sensitivity and specificity of combined evaluation of severe acne were 90.91% and 71.43%, respectively. Conclusion The level of 25(OH) D is decreased and the level of IGF-1 is increased in patients with acne, which is related to the severity of the disease. The combination of 25(OH) D and IGF-1 levels has a better efficacy in evaluating severity acne.

Abstract: Objective　To construct HepG2, Huh7 cell lines stably express hepatitis B virus X (HBx) mutant (C1653T, T1753C), and explore their effect on the biological behavior of hepatocellular carcinoma cells. Methods　The lentivirus plasmid of pLVX-HBxC1653T-IRES-tdTomato, pLVX-HBxT1753C-IRES-tdTomato were obtained by PCR site mutagenesis according to wild type ayr HBx. Double enzyme digestion and Sanger sequencing were performed for accuracy of plasmid. Blank HepG2 and Huh7 cells were used as the control group, HepG2, Huh7 cells were infected by pLVX-HBx-IRES-tdTomato, pLVX-HBxC1653T-IRES-tdTomato, and pLVX-HBxT1753C-IRES-tdTomato lentivirus solution, then monoclonal cell was selected by 0.6 μg/mL puromycin. Immunostaining and Western Blot were performed for the verification of stable strains. CCK8 assay was performed for the proliferation capacity of stable strains. Western Blot was performed for expression of EMT-related signal molecules in cells. The independent samples t-test was used for comparison between two groups. Results Double enzyme digestion and Sanger sequencing showed that that the size of the cut fragments of recombinant lentiviral plasmids was correct, and the point mutation location and base substitution were correct, suggesting that the plasmid of pLVX-HBx-IRES-tdTomato, pLVX-HBxC1653T-IRES-tdTomato, pLVX-HBxT1753C-IRES-tdTomato were constructed successfully. Immunostaining and Western blot showed that HBX were expressed in stable strains, while there was no HBX expression in the blank control group, indicating that the HepG2 and Huh7 cell lines stably expressing HBx, HBxC1653T, HBxT1753C were successfully constructed. CCK8 assay showed that the proliferation capacity of HBx and mutant were enhanced compared to the control group (P<0.01), HBx C1653T displayed further additive the effect compared to HBx (P<0.05). Moreover, HBxC1653T mutation also significantly upregulated N-cadherin expression and downregulated E-cadherin expression, thus promoting the occurrence of EMT. Conclusions　HepG2 and Huh7 cell lines stably expressing HBx, HBxC1653T, HBxT1753C were successfully constructed, HBxC1653T mutation significantly enhanced the proliferation of HCC cells and epithelial to mesenchymal transition occurrence.

ObjectiveTo explore the mechanism of Si Junzitang in regulating the expression of NKG2A to affect the anti-colon cancer function of natural killer （NK） cells. MethodNK cells isolated from healthy honors were cultured and used to construct the three incubation models of NK cells， human colon cancer HCT116 cells， and NK cells + HCT116 cells （co-incubation）. real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） was conducted to determine the mRNA levels of natural killer group 2 member A （NKG2A） and interleukin （IL）-15 in NK cells， as well as the mRNA level of histocompatibility leucocyte antigen E （HLA-E） in HCT116 cells. The secretion of IL-15 was detected by enzyme-linked immunosorbent assay （ELISA）. Methyl thiazolyl tetrazolium （MTT） assay was employed to determine the applicable concentration of IL-15 and test the effects of Si Junzitang and IL-15 on the activities of NK cells and the HCT116 cells in the co-incubation model. The effects of Si Junzitang and IL-15 on the mRNA levels of NKG2A in NK cells and HLA-E in HCT116 cells were detected by Real-time PCR. Monalizumab （M， anti-NKG2A mab） was used to block the NKG2A-HLA-E pathway in co-incubation model， and then the proliferation of HCT116 cells was detected by MTT assay. ResultThe interaction of NK cells and HCT116 cells up-regulated the mRNA levels of NKG2A in NK cells and HLA-E in HCT116 cells （P<0.05）， as well as the expression level and secretion of IL-15 （P<0.05）. Compared with the blank group， Si Junzitang and Si Junzitang + IL-15 promoted the proliferation and improved the anti-colon cancer function of NK cells （P<0.01）. Furthermore， they down-regulated the mRNA levels of NKG2A in NK cells and HLA-E in the HCT116 cells co-incubated with NK cells （P<0.01）. M and IL-15 + M inhibited the proliferation of HCT116 cells compared with the groups without M （P<0.01）. ConclusionThe interaction of NK cells and HCT116 cells can induce activation of NKG2A-HLA-E pathway to impair NK cell function. Si Junzitang can inhibit the activation of NKG2A-HLA-E pathway to restore the anti-colon cancer function of NK cells.

Objective:To establish the quality evaluation method of Prunellae spica dispensing granules based on three quality indexes of standard decoction. Methods:Fourteen batches of Prunellae spica were collected from different habitats. According to technical requirements, fourteen batches of Prunellae spica standard decoction and three batches of formula granules were prepared and the paste-forming rates were calculated. The fingerprints of Prunellae spica standard decoction and formula granules were established by Ultra High Performance Liquid Chromatography (UPLC). The similarity values of fingerprints between dispensing granules and standard decoction were calculated. The content and transferring rate of Rosmarinic acid were determined and calculated. Results:The average paste-forming rate of Prunellae spica was (12.59±2.32)%. The paste-forming rates of the three batches were 11.14%, 10.78% and 10.39% respectively. The average content of Rosmarinic acid in standard decoction was (18.99±9.74)mg/g. The average transferring rate was (60.58±7.87)%. The contents of three batches were 7.40 mg/g, 7.49 mg/g and 7.09 mg/g. The transferring rates were 52.06%, 50.10% and 50.40% respectively. Nine common fingerprint peaks were identified in the fingerprints of standard decoction and formula granules, two of which were identified as Rosmarinic acid and Caffeic acid by comparison of reference substance. The fingerprints similarity of Prunellae spica dispensing granules and standard decoction were 0.954, 0.973 and 0.952, respectively. Conclusions:The quality indexes of three batches of formulation granules are consistent with standard decoction. This method could provide reference for the establishment of quality standard of Prunellae spica dispensing granules.

Rotundic acid(RA),an ursane-type pentacyclic triterpene acid isolated from the dried barks of Ilex rotunda Thunb.(Aquifoliaceae),possesses diverse bioactivities.To further study its pharmacokinetics,a simple and sensitive liquid chromatography with triple quadrupole mass spectrometry(LC-QqQ-MS/MS)method was developed and validated to quantify RA concentration in rat plasma and tissue using etofesalamide as an internal standard(IS).Plasma and tissue samples were subjected to one-step protein precipitation.Chromatographic separation was achieved on a ZORBAX Eclipse XDB-C18 col-umn(4.6 mm×50 mm,5 μm)under gradient conditions with eluents of methanol:acetonitrile(1∶1,V/V)and 5mM ammonium formate:methanol(9∶1,V/V)at 0.5mL/min.Multiple reaction monitoring transitions were performed at m/z 487.30 → 437.30 for RA and m/z 256.10 → 227.10 for IS in the negative mode.The developed LC-QqQ-MS/MS method exhibited good linearity(2-500 ng/mL)and was fully validated in accordance with U.S.Food and Drug Administration bioanalytical guidelines.Dose proportionality and bioavailability in rats were determined by comparing pharmacokinetic data after single oral(10,20,and 40 mg/kg)and intravenous(10 mg/kg)administration of RA.Tissue distribution was studied following oral administration at 20 mg/kg.The results showed that the absolute bioavailability of RA after administration at different doses ranged from 16.1％to 19.4％.RA showed good dose proportionality over a dose range of 10-40 mg/kg.RA was rapidly absorbed in a dose-dependent manner and highly distributed in the liver.In conclusion,this study is the first to systematically elucidate the absorption and distribution characteristics of RA in rats,which can provide additional information for further development and evaluation of RA in drug metabolism and pharmacokinetic studies.

Objective:To investigate the clinical features and perinatal outcomes of twin pregnancies with complete placenta previa (CPP).Methods:We conducted a retrospective study on 266 women with CPP, including 62 twin pregnancies (twins group) and 204 singleton pregnancies (singleton group), who gave birth in Peking University Third Hospital from January 2012 to December 2020. T-test, nonparametric test and Chi-square test were adopted for univariate analysis. Differences between the two groups regarding clinical features and perinatal outcomes were compared using multivariate logistic regression or multivariate linear regression. Results:The incidence of twin pregnancy with CPP was 2.11% (62/2 937). Placenta accreta spectrum disorders (PAS) accounted for 48.4% (30/62) and 53.9% (110/204) in the twin and singleton group, respectively, but the difference was not statistically significant ( χ 2=0.58, P>0.05). In terms of antepartum hemorrhage, the proportion of women affected, those with first onset <29 weeks, amount of bleeding ≥200 ml, and the number of episodes of bleeding ≥3 were significantly higher in the twin group than those in the singletons [56.5% (35/62) vs 39.7% (81/204); 35.5% (22/62) vs 12.7% (26/204); 17.7% (11/62) vs 4.9% (10/204); and 21.0% (13/62) vs 10.3% (21/204), χ 2=5.42, 16.62, 10.78, and 4.86, respectively, all P<0.05]. Multivariate Logistic regression analysis showed that compared with the singleton group, the twin group was at higher risk of antepartum hemorrhage volume >200 ml, the number of antepartum hemorrhage episodes ≥3, preterm delivery before 34 weeks and 32 weeks, emergency cesarean section, and emergency cesarean section caused by antepartum hemorrhage [a OR(95% CI)=4.36(1.17-16.30), 3.15(1.01-9.79), 17.24(5.36-55.46), 9.85(2.32-41.77), 3.98(1.72-9.20), and 3.10(1.22-7.85), respectively, all P<0.05]. Multivariate linear regression analysis showed that the gestational week at the emergency cesarean section in the twins group was about 2.22 weeks (0.17-4.27 weeks) earlier than that in the singletons. The postpartum hemorrhage amount and the risk of postpartum hemorrhage after cesarean section, infusion of red blood cells, and hysterectomy did not differ significantly between the two groups. Conclusions:Compared with singleton pregnancies, the proportion of preterm delivery, cesarean sections, especially those caused by antepartum hemorrhage, is significantly higher among twin pregnancies combined with CPP. Accordingly, preterm delivery should be actively prevented, and the timing of cesarean section should be individualized according to the condition of the mothers and babies, and early delivery may be considered.

Objective:Postoperative venous obstruction (PVO) is the most severe complication of total anomalous pulmonary venous connection (TAPVC), and facing challenging re-intervention with high mortality. We aimed to review and analyze the follow-up and management of postoperative PVO in our center.Methods:We conducted a retrospective study of the patients with isolated TAPVC admitted in our center from October 2013 to October 2019. All available data and images of PVO patients were reviewed, such as the initial perioperative medical records, patients’ follow-up records, results of patients’ echo and CT angiography. Re-intervention including hybrid technique, sutureless technique, and patch augmentation, were carried out for postoperative PVO patients. The results were reviewed and analyzed to find the risk factors for adverse prognosis.Results:A series of 174 isolated TAPVC patients were admitted in our center and 169 received surgical treatment and 26 (26/169, 15.4%) had postoperative PVO. The diagnosis was made at a median time of 11.5 (0-77) weeks after initial operation and within 6 months of surgery in 22 (22/26, 84.6%) of the 26 patients. The subtype of TAPVC patients with postoperative PVO were: supracardiac 11 cases (11/26, 42.3%), cardiac 7 cases (7/26, 26.9%), infracardiac 5 cases (5/26, 19.2%), and mixed 3 cases (3/26, 11.5%). Bilateral obstruction and stenosis with diffusely small pulmonary veins were in 12 (12/26, 46.2%) and 3 cases (3/26, 11.5%) respectively. PVO progressed to worse condition in all the 26 cases during follow-up period. 8 (8/26, 30.8%) postoperative PVO patients underwent 10 re-interventions: one cases had 3 re-interventions. Five-year survival for patients with postoperative PVO was worse than those without postoperative PVO ( HR=6.46, 95% CI: 2.34-17.85, P<0.01). Risk factors for death or re-intervention in postoperative PVO patients were earlier presentation after TAPVC repair ( HR=0.85, 95% CI: 0.73-0.99, P=0.04) and an increased number of lung segments affected by obstruction ( HR=1.74, 95% CI: 1.01-2.99, P=0.04). Conclusion:Risk factors for death or re-intervention in postoperative PVO patients were earlier presentation after TAPVC repair and an increased number of lung segments affected, which should be focused on during strict follow-up period. Early re-intervention should be taken before irreversible secondary changes occur in these patients.