ObjectiveTo investigate the mechanism by which Notoginsenoside R1 （NGR1） ameliorates hypoxia/reoxygenation （H/R）-induced injury in AC16 human cardiomyocyte cell lines through the regulation of mitophagy. MethodsCommon genes linked to hypoxia/reoxygenation injury and mitophagy were identified by intersecting data from GeneCards and MitoCarta databases. AC16 cell viability was assessed via CCK-8 assay under varying NGR1 concentrations （0， 6.25， 12.5， 25， 50， 100， 200， 300， 400， 500 μmol/L）. AC16 cells were divided into the following groups： control group （Control）， model group （H/R）， and treatment groups （H/R + NGR1 at 100， 200 and 300 μmol/L）. Mitochondrial membrane potential （ΔΨm） was measured using 5，5'，6，6'-tetrachloro-1，1'，3，3'-tetraethylbenzimidazolylcarbocyanine iodide （JC-1） staining. Transcriptional levels of mitophagy-related genes （Parkin， Pink1， P62） were quantified by reverse transcription-quantitative PCR （RT-qPCR）. Protein expression of mitophagy-related markers （Parkin， Pink1， P62， and LC3BⅡ） was evaluated via Western blot analysis. Mitochondrial ultrastructure was visualized by transmission electron microscopy （TEM）. ResultsCompared to the control group， cell viability in the H/R group significantly decreased （P<0.01）. Treatment with NGR1 at concentrations above 100 μmol/L significantly enhanced the cell viability of AC16 cells compared to the H/R group （P<0.01）. H/R induced a significant decrease in mitochondrial membrane potential （P<0.01）， which was restored by NGR1 treatment （P<0.01）. The mRNA levels of Parkin， Pink1， and P62 in the H/R group were upregulated compared to the control group （P<0.05）， while NGR1 intervention downregulated their expression （P<0.05）. Protein expression levels of Parkin， Pink1， and LC3BⅡ in the H/R group significantly increased， while P62 expression decreased compared to the control group （P<0.01）. In contrast， different doses of NGR1 treatment significantly reduced the expression of Parkin， Pink1， and LC3BⅡ while increasing P62 expression （P<0.05）. TEM revealed that the mitochondrial structure in the H/R group was severely disrupted， with fragmented and disorganized cristae， which was alleviated by NGR1. ConclusionNGR1 ameliorates H/R-induced AC16 cell injury， and its mechanism may be associated with modulating the Pink1/Parkin pathway to suppress excessive mitophagy.

ObjectiveThe U6 promoter is an essential element for driving sgRNA expression in the clustered regularly interspaced short palindromic repeat sequences/CRISPR-associated protein 9（CRISPR/Cas9）gene editing system in dicotyledonous plants. Endogenous U6 promoters typically exhibit higher transcriptional activity， which can significantly improve gene editing efficiency. This study aims to identify endogenous U6 promoters in Artemisia annua to optimize its CRISPR/Cas9 gene editing system， which holds significant importance for its molecular breeding. MethodsOn the basis of the highly conserved U6 snRNA sequences in Arabidopsis thaliana， endogenous U6 promoters were screened in the A. annua genome. Expression vectors were constructed with candidate AaU6 promoter driving the firefly luciferase （LUC） reporter gene， and then transiently transformed into Nicotiana benthamiana. Transcriptional activities of the promoters were measured and compared by in vivo imaging and the Dual Luciferase Reporter assay. ResultsEight endogenous U6 promoters were successfully cloned from A. annua. Sequences alignment revealed that all these promoters contained the two conserved cis-acting elements， upstream sequence element （USE） and TATA-box， which affected their transcriptional activity. Dual-luciferase activity assays indicated that the transcriptional activities of AaU6-3， AaU6-1， and AaU6-5 were significantly higher than that of the Arabidopsis AtU6-26 promoter， with AaU6-3 exhibiting the highest activity. ConclusionThis study identified three endogenous AaU6 promoters with high transcriptional activity in A. annua， providing key functional elements for establishing an efficient gene editing system in A. annua. These findings will contribute to advancing precision molecular breeding and high-quality germplasm innovation in A. annua.

Objective Explore the impact of long-term blood glucose fluctuations and blood glucose control on the stroke incidence in patients with type 2 diabetes mellitus(T2DM).Methods An observa-tional cohort of patients with T2DM was established based on"Shanghai Community Chronic Disease Health Management Service Objects"on October 1st,2018.Follow-ups were conducted every three months,and fasting blood glucose(FBG)were tested at each visit.Basic epidemiological data were collected via the Shanghai Community Health Management Information Platform or survey questionnaires,and stroke incidents were gathered via the"Shanghai Cardio-Cerebrovascular Event Monitoring System".The first reported stroke incident within the observation period was considered.The observational deadline was December 31st,2021.Standard deviation of FBG was used to evaluate blood glucose fluctuation and FBG control rate was used to reflect blood glucose control status.Cox Proportional Hazards Model was utilized to analyze the impacts.Results The cumulative observation time was 91,826.1 person-years for the study,in which there were 1785 cases of stroke events observed,implying a cumulative incidence of 5.73%and incidence rate of 1943.9/100000 person-years.The mean number of follow-ups(10.29±3.07)per patient was recorded.The details of the stroke cases were as follows:transient ischemic attacks(n=111,6.22%),non-lacunar brain infarctions(n=754,42.24%),lacunar brain infarctions(n=798,44.71%),intracerebral hemorrhages(n=80,4.48%),non-intracerebral hemorrhages(n=8,0.45%),subarachnoid hemorrhages(n=12,0.67%),and unclassified strokes(n=22,1.23%).After excluding subjects with less than five follow-ups,the Cox Proportional Hazards Model suggested that increased standard deviation of FBG was an independent risk factor for the first stroke incident,and an increased blood glucose control rate was an independent protective factor.Both the standard deviation of FBG and the blood glucose control rate were not correlated with stroke recurrence.Conclusion Long-term blood glucose fluctuations and blood glucose control can predict the risk of a first stroke in patients with T2DM.The relationship between blood glucose fluctuations,blood glucose management,and stroke recurrence requires further studies.

【Objective】 To analyze the correlation between CT attenuation value of renal papilla and occurrence and development of renal calculi. 【Methods】 The clinical data of 100 patients with calcium oxalate stones treated during Aug.2020 and Jul.2022 were retrospectively analyzed, including 60 with primary stones, and 40 with recurrent stones. The 30 healthy volunteers were enrolled in the control group. CT attenuation value of renal papilla was measured with plain scan CT, and risk factors were identified with the receiver operating characteristic (ROC) curve. The correlation of CT attenuation value and 24 h urine metabolism was analyzed. 【Results】 The CT attenuation value of renal papilla was significantly higher in the primary group [34.92 (IQR: 3.84)] and recurrent group [43.00 (IQR: 8.74)] than in the control group [32.58 (IQR: 5.21)] (P<0.05). Compared with the primary group, the recurrent group had decreased citric acid level but increased calcium level in 24 h urine (P<0.05). The citrate ion and calcium ion were correlated with the CT attenuation value (P<0.05). 【Conclusion】 Patients with high renal papilla density have a high risk of stone formation and recurrence. Increased renal papilla density is a warning signal for the development of stones. The high calcium and low citric acid in 24 h urine have certain effects on the occurrence and development of urolithiasis. Intake of calcium should be limited and citric acid should be supplemented in patients with calcium oxalate stones.

Objective:To investigate the 2 years’ efficacy of intravesical instillation of domestic BCG versus epirubicin in the prevention of recurrence of intermediate-risk or high-risk non-muscular invasive bladder cancer and predictive factors of BCG instillation.Methods:From July 2015 to June 2020, 18-75 years old patients with moderate to high-risk non muscle invasive bladder cancer (NMIBC) confirmed by pathological examination were involved. The ECOG score was 0-2. Exclusion criteria included ①immune deficiency or impairment (such as AIDS), using immunosuppressive drugs or radiotherapy, suspected allergic to BCG or epirubicin or excipients of the two drugs, fever or acute infectious diseases including active tuberculosis or receiving anti tuberculosis treatment, with severe chronic cardiovascular and cerebrovascular diseases or chronic kidney disease; ②combined with other urogenital system tumors or other organ tumors; ③combined with muscle invasive bladder urothelial carcinoma (≥T 2); ④undergoing chemotherapy, radiotherapy or immunotherapy within 4 weeks (immediate instillation after surgery not included); ⑤ pregnant or lactating women; ⑥ comfirmed or suspected bladder perforation; ⑦gross hematuria; ⑧cystitis with severe bladder irritation that may affect the evaluation; ⑨participat in other clinical trials within 3 months; ⑩alcohol or drug addiction; ?any risk factors that may increasing the risk of patients. Epirubicin 50 mg was irrigated immediately after the operation(TURBT or laser resection). The patients were randomly divided into BCG15 group, BCG19 group and epirubicin group by the ratio of 2∶2∶1, and the patients were maintained intravescical instillation for 1 year. The recurrence and adverse events of the three groups were compared. Univariate and multivariate analysis was performed to predict the risk factors of BCG irrigated therapy failure. Result:By June 15, 2020, the median follow-up duration was 22.1 months(12.1, 32.3), and there was no statistical difference between the groups ( P=0.9024). There were 274 patients enrolled in BCG19 group, 277 patients enrolled in BCG15 group and 130 patients enrolled in the epirubicin group. The drop-off rate was 16.6%(113 cases)and made no difference between groups( P=0.6222). There were no significant difference in age, gender, BMI, or ECOG score( P>0.05). During the follow-up, 116 cases was detected recurrence or progression. The recurrence rate of the three groups was 14.2% and 14.8% in BCG19 group and BCG15 group, and 27.7% in the epirubicin group. There was no difference in recurrence rate between BCG19 and BCG15 group( P=0.9464). The recurrence rate of BCG19 group was lower than that of the epirubicin group ( P=0.0017). The recurrence rate of BCG15 group was lower than that of the epirubicin group ( P=0.0020). There was no difference in the cumulative recurrence free survival rate between BCG19 and BCG15 group (95% CI0.57-1.46, P=0.7173). The cumulative recurrence free survival rate of BCG 19 group was better than that of the epirubicin group( HR=0.439, 95% CI0.26-0.74, P=0.0006), and the cumulative recurrence free survival rate of BCG15 group was better than that of the epirubicin group ( HR=0.448, 95% CI0.29-0.80, P=0.0021). The total incidence of adverse events in 19 BCG19, BCG15 and epirubicin group were 74.5%, 72.6% and 69.8% respectively. There was no difference in the incidence of adverse events between BCG19 and BCG15 group( P=0.6153). The incidence of adverse events in epirubicin group was lower than that of BCG19( P=0.0051) and BCG15( P=0.0167) groups.There was no significant difference in the incidence of serious adverse events (SAE) among the three groups ( P=0.5064). Log rank test univariate analysis and Cox risk regression model multivariate analysis showed that the history of bladder cancer recurrence( HR=6.397, 95% CI1.95-20.94, P=0.0001)was independent risk factor for BCG irrigation failure. Conclusions:The 2 years’ efficacy of intravesical instillation of domestic BCG is better than than of epirubicin with good tolerance and safety. There is no difference between BCG19 and BCG15 group. BCG doesn’t increase SAE compared with epirubicin. Recurrence status was an independent prognostic factor regarding recurrence-free survival.

Objective: To analyze the change trend of HIV genetic subtypes and compare the first CD₄⁺T cell counts of newly diagnosed HIV infected patients in Liuzhou from 1998 to 2012, and provide a reference for AIDS prevention and control. Methods: Newly diagnosed HIV-infected patients from 1998 to 2012 in Liuzhou were selected through national HIV/ADIS comprehensive response information management system. Their plasma samples were used for RNA gene extraction, amplification, sequencing and genotyping. Coharan-Armitage trend test was used to analyze the ratio trend of genetic subtypes and phylogenetic clusters of HIV and Wilcoxon Rank Sum Test was used to compare the first CD₄⁺T cell counts (CD₄) of the different subtype HIV infected patients. Results: A total of 1 877 newly diagnosed HIV infected patients were included in the study. From 1998 to 2012, the proportions of CRF01_AE and CRF01_AE (Cluster 1) increased from 78.4% (76/97) to 91.5% (1 441/1 574), from 63.9% (62/97) to 74.0% (1 164/1 574), and the proportion of CRF07_BC decreased from 17.5% (17/97) to 4.6% (72/1 574), respectively (Z=4.632, P<0.001; Z=2.455, P=0.014; Z=-5.943, P<0.001). The median and interquartile range of the first CD₄ of the patients infected with subtype CRF01_AE (Cluster 1), CRF01_AE (Cluster 2), CRF07_BC and CRF08_BC were 230 (83-375), 215 (48-351), 365 (254-503) and 334 (206-479) cell/μl, respectively. The first CD₄ levels of the patients infected with subtype CRF01_AE (Cluster 1) or CRF01_AE (Cluster 2) were significantly lower than those of CRF07_BC (Z=-4.795, P<0.001; Z=-4.238, P<0.001). Conclusion The genetic subtypes of HIV were mainly CRF01_AE in newly diagnosed HIV-infected patients and this subtype proportion was in increase and the first CD₄ levels of the patients were low in Liuzhou during 1998 to 2012.

The annual congress of Japan Surgical Society is a famous academic event in the field of surgery,and the participants can learn from the latest research results of all the major surgical disciplines.The authors selected topics of the ll8th annual congress in 2018,including the latest research results and progresses of laparoscopic hepatectony,liver transplantation,extrahepatic bile duct carcinoma and pancreatic surgery.The purpose of this study is to provide new information and reference for optimizing the diagnosis and treatment of hepatobiliary and pancreatic diseases.

Objective To investigate the effect of Shensong Yangxin capsule on cardiac remodelling of myocardial infarction mouse model and the possible molecular mechanisms. Methods Adult male C57BL/6J mice were divided into sham operation group(n=10), model control group(n=20)and Shensong Yangxin group(n=20)according to random number table. Left anterior descending branch of coronary artery was ligated to establish myocardic infarction model in the model control group and Shensong Yangxin group. From the 2nd day after the surgery, Shensong Yangxin ( 400 mg . kg-1 ) was intragastrically administered, and the death rate of the mice was observed.Four weeks after the surgery, echocardiography was used to measure the cardiac function;myocardiac infarction area was detected by pathological staining;the expression levels of cardiac remodelling markers and extracellular matrix proteins were detected by RT-PCR. The possible molecular mechanisms were screened by Western blotting. Results As compared with the model control group, Shensong Yangxin significantly reduced the mortality after myocardial infarction in mice(P<0.05), as well as the myocardial infarct size(P<0.05).The mRNA expression levels of cardiac remodelling markers ANP, BNP, and β-MHC and the extracellular matrix proteins(collagenⅠ, collagen Ⅲ, CTGF, TGFβ) decreased significantly in the Shensong Yangxin group as compared with the model control group. Western blotting showed that Shensong Yangxin significantly decreased activation of smad3, and reduced expression level of smad4. Conclusion Shensong Yangxin attenuates cardiac remodelling after myocardial infarction and the mechanism may be related with blockage of smad signaling pathway.

Objective To explore the influence of ozone therapy on high-sensitivity C-reactive protein (hs-CRP) and matrix metalloproteinases-3 (MMP-3) in synovial fluid of patients with knee osteoarthritis,and to investigate the possible mechanism.Methods Sixty-two patients with knee osteoarthritis were divided into observation group and control group by random digits table method with 31 cases each.The patients of observation group were treated with ozone,while the patients of control group were treated with hyaluronic acid once a week for 5 weeks.The efficacy was evaluated and the level of hs-CRP and MMP-3 in synovial fluid were detected before and after treatment.Results The total effective rate in observation group was 100.00％ (31/31),significantly higher than that in control group [87.10％ (27/31)],and there was significant difference (x2 =4.28,P ＜ 0.05).After treatment,the level of hs-CRP and MMP-3 in observation group was (6.68 ±2.15) mg/L,(69.84 ± 16.97) μ g/L,significantly lower than that in control group [(8.72 ± 2.39) mg/L,(85.43 ± 19.52) tμ g/L],and there was significant difference (t =3.53,3.36,P ＜ 0.01).Conclusion There is significant effect of ozone therapy of patients with knee osteoarthritis,its mechanism may be reducing the level of MMP-3 and inhibiting inflammatory reaction.

Objective To investigate the causes of complications induced by peripherally inserted central catheters(PICC)in cancer patients and summarize pertinent nursing measures.Method The clinical data of 26 tumor patients with PICC were analyzed retrospectively .Result The complications occurred in 8 patients with a rate of 30.8%, among which unsuccessful catheteration occurred in 1 case,oozing of blood at the mouth of catheter in 1 case,phlebitis in 1 case,dislocation in 1 case,skin allergy-like reactions in 2 cases and catheter emersion in 1 case,catheter breaking in 1 case and dislocation in 2 cases.Conclusion The skilled operation,preventive maintenance of catheters and health education to the patients are the important measures for prevention and reduction of the complications.