Objective To investigate the situation of HIV infection and epidemiological characteristics among voluntary blood donors and pre-transfusion patients in Yuxi prefecture between 2010 and 2021.Methods Da-te collected with the HIV/AIDS Case Reporting Cards and original record of blood station,HIV antibody positive rate,the demographic characteristics,and epidemiological data of blood donors and pre-transfusion patients,were analyzed accordingly,the related date were statistically analyzed.Results The HIV antibody positive ratio were 5.56? and 2.01? among pre-transfusion patients and voluntary blood donors(P<0.001),showing an downward trend year after year.The HIV-positive detection rate among blood donors aged<25 years was significantly higher at 3.72‰ compared to 1.60‰ for those aged≥25 years(P=0.003).Among HIV-positive blood recipients,the proportion of individuals aged 15-24 years(34.62%)was markedly higher than that among blood donors(1.96%),whereas no donors were aged≥50 years compared to 23.53%of recipients(P<0.001).Farmers showed a significantly higher HIV detection rate(3.18‰)than students(2.66‰)and other occupations(1.60‰)(P=0.041).Similarly,individuals with high school education or below showed a higher detection rate(2.52‰)than those with high school education or above(1.45‰)(P=0.045).Among HIV-positive cases,blood donors had a higher proportion of HIV-infected individuals(84.62%)versus AIDS cases(15.38%),whereas the recipients showed the opposite trend:41.18%HIV-infected versus 58.82%AIDS cases(P<0.001).Conclusion In recent years,the HIV positive detection rate in Yuxi area has shown a downward trend,but the occurrence of HIV in Yuxi area is characterized by youthfulness and low education.Among them,HIV infected individuals through same-sex contact account for an important reason for HIV infection.

Objective:To investigate the characteristics of spatiotemporal heterogeneity in pathological grading during the latent and invas-ive growth phases of non-metastatic renal cell carcinoma(RCC)and its correlation with clinical outcomes.Methods:A retrospective analysis was conducted on the case data of 316 RCC patients with local recurrence(LR)and 429 RCC patients with venous tumor thrombus(VTT)who underwent surgical treatment at 13 medical centers in China from January 2003 to December 2023.Pathological grade differences between primary tumor(PT)and LR,and between PT and VTT were selected as scenarios for the application of spatiotemporal heterogen-eity in the dynamic progression of RCC.Pathological grading changes were defined according to a new four-tier scheme(upgrading,down-grading,stable low-grade,and stable high-grade).Stable low-or high-grade was defined as low-grade(WHO/ISUP grade Ⅰ or Ⅱ)or high-grade(WHO/ISUP grade Ⅲ or Ⅳ)in both PT and LR/VTT.Upgrading was defined as low-grade in the PT and high-grade in the LR/VTT;con-versely,downgrading was defined as high-grade in the PT and low-grade in the LR/VTT.The potential influencing factors of pathological grading changes and their impact on patient prognosis were analyzed.Results:The median cancer-specific survival(CSS)for RCC patients with VTT and RCC patients with LR was 83 months and 76 months,respectively.The 5-year CSS rates were 65.6%and 60.6%,respectively.Pathological grading changes were observed in 38.0%of patients with PT and VTT and in 43.6%of patients with PT and LR.Lasso-Logistic re-gression analysis revealed a close correlation between primary tumor necrosis and pathological grading changes.Kaplan-Meier survival curves indicated a significant correlation between pathological grading changes and prognosis.Replacing the pathological grading in Leibovich,UISS,and GRANT scores with pathological grading changes significantly improved the predictive performance of the models(P<0.05).Conclusions:Spatiotemporal heterogeneity in pathological grading exists during the dynamic progression of non-metastatic RCC.Compared to the pathological grading of isolated events,the spatiotemporal variation in pathological grading serves as a more accurate in-dependent prognostic factor for RCC patients with VTT and RCC patients with LR.Incorporating pathological grading changes can signific-antly improve the predictive performance of existing prognostic models.

Objective:High-throughput screening to obtain small molecular compounds against Gram-negative bacilli by targeting BamA outer membrane protein.Methods:The sybyl-X2.1 software was used to perform high-throughput virtual screening of small molecular compounds in Chemdiv compound library based on the molecular docking. The top 150 hits by high-throughput screening were re-screened through in vitro biological experiments. The top 4 small molecules with obvious antibacterial activity were selected for in-depth molecular docking analysis, and the small molecule 8308-0401 with the highest docking score was selected for further experiments. The antibacterial effect of 8308-0401 combined with rifampicin was tested by checkerboard assay. Finally, the affinity between 8308-0401 and BamA was tested by plasma surface resonance assay. Results:The docking score of the top 150 hits calculated by high-throughput virtual screening had a mean value of 5.63. In vitro biological experiments showed that small molecules 8308-0401, 8365-1335, C066-2507 and L582-0346 exhibited strong antibacterial activity. Among those molecules, 8308-0401 showed the highest molecular docking score, and synergistic antibacterial activity against both types of strains and clinical isolates when combined with rifampicin. 8308-0401 has a strong affinity to BamA with binding a constant of 182 μmol/L. Conclusion:The small molecule 8308-0401 exerts antibacterial activity against Gram negative bacilli by targeting the outer membrane protein BamA.

Objective:To investigate the feasibility and safety of a novel single-port robotic surgical system with flexible 3D endoscope and deformable surgical instruments in zero ischemic partial nephrectomy.Methods:From May 2021 to October 2021, a prospective study on patients with renal tumor who plan to receive zero ischemic partial nephrectomy was conducted. Inclusion criteria included over 18 years old, body index between 18.5 to 30.0 kg/m2, American Society of Anesthesiologists Score ranged from 1 to 3 points, cooperation with the follow-up and related examinations, voluntary in participating the clinical trial and signing the informed consent. Exclusion criteria included patients with other co-existed malignant tumors or a medical history of other malignant tumors, the patients who have received the same urological surgery in the past, the patients who underwent or plan to undergo other major operations 3 months before or after the surgery, the patients with active pulmonary tuberculosis or severe systemic diseases, the patients to be considered not suitable to enroll in by the researchers. A novel single-port robotic surgical system was used to perform the surgery. The system consiststed of a remote control console, a surgical equipment cart, a four-arm operation cart and deformable robotic instruments which were reusable. The two-section deformable robotic instruments were able to bend in four directions and carried different surgical instruments such as unipolar scissors, bipolar grasping forceps and needle holders. The deformable robotic instruments entered the body through a special trocar with single hole and multi-channel, and then unfolded in a triangle. By bending instruments, surgeons could perform single-port robotic surgery without the trouble of "chopstick effect" or "reverse direction" . Four cases of single-port partial nephrectomy were carried out. Under general anesthesia, the patients were taken the lateral recumbent position, with elevated waist and lowered head and feet. A 3-4 cm incision was taken layer by layer along the lateral edge of the rectus abdominis at the umbilicus level. A special 2.5 cm robotic trocar was set into the cavity, and a high-definition 3D laparoscopic lens and a snake shaped mechanical arm were then put into the abdominal cavity through the trocar. All operations were performed by transperitoneal approach with an auxiliary port through the same or a different skin incision if necessary. Tumor resection and renal reconstruction were performed by the way of zero ischemia. The perioperative parameters such as tumor size, operation time, intraoperative bleeding and postoperative complications were analyzed.Results:Four patients were involved, including 2 males and 2 females, with 2 of them having a history of hypertension. The ECOG scores were all 0, and KPS score was 100 in 3 cases and 90 in 1 case. Preoperative mean serum creatinine was (76.8±18.8)μmol/L (range 70-104 μmol/L). The tumors were located on the left in 3 cases and on the right in 1 case. The diameter of the tumor ranged from 1.1 to 2.8 cm, with the TNM classification of T 1a. The R. E.N.A.L. scores were 4a, 7p, 6p and 4P respectively. The first operation was performed by pure single-port surgery, and the other 3 cases were performed with the help of an auxiliary port to ensure the safety .The operation time ranged from 155 min to 210 min, and the intraoperative bleeding ranged from 20 ml to 170 ml. Postoperative pathology showed 2 cases of renal clear cell carcinoma with negative margin and 2 cases of angiomyolipoma. No severe complications, such as bleeding or urinary leakage, were observed during the perioperative period, and the change of serum creatinine was insignificant before discharge and before operation( P=0.24). Conclusions:A robotic single-port partial nephrectomy can be successfully carried out by using this novel single-port robotic surgical system with flexible 3D endoscope and deformable surgical instruments.

Objective:To explore the surgical technique and efficacy of pure retroperitoneoscopic extravesical standardized seeable (P.R.E.S.S.) technique for bladder cuff excision (BCE).Methods:Ninety five patients with UTUC from five domestic centers (30 cases from Changzheng Hospital, 21 cases from Peking University First Hospital, 20 cases from Yuhuangding Hospital, 21 cases from Dalian Medical University affiliated No.2 Hospital and 3 cases from General Hospital of Eastern Theater Command)between August 2017 and December 2020 were retrospectively analyzed. There were 57 males and 38 females with a mean age of (67.7±10.0) years and median tumor size of 3.0 cm. All patients underwent pure retroperitoneoscopic radical nephroureterectomy with a single surgical position and four (36 cases) or five (59 cases) trocar layout according to the surgeon’s prefer habit and experience. The demographics of the two groups were the age of [(66.3±11.2)years vs. (68.6±9.1)years], male/female ratio of (25/11 cases vs. 32/27 cases), body mass index of [(25.0± 3.0)kg/m 2 vs. (24.8±3.4)kg/m 2], tumor maximum diameter of [2.8(1.6, 3.5)cm vs. 3.0(2.0, 4.0)cm], left/right side tumor of(19/17 cases vs. 34/25 cases), T 1-2/T 3-4/Tis stage of(25/10/4 cases vs. 49/10/0 cases), and multifocal tumors of(3 cases vs. 2 cases), and the difference was not statistically significant( P>0.05). On the other hand, the differences of hydronephrosis of the operated kidney(13 cases vs. 39 cases, P=0.004), and tumor location (in renal pelvis or calyx or upper/middle/lower ureter being 23/9/4 cases vs. 35/4/20 cases, P=0.005), were statistically significant. The umbilical artery cord was used as anatomical landmark in the process of P. R.E.S.S. bladder cuff excision. The pelvic floor and extraperitoneal space around the ureter were expanded, the bladder wall was opened to form pneumovesicum, and a sufficient bladder cuff resection and exact bladder cuff closure was performed. Perioperative outcomes and follow-up data were analyzed, and the clinical outcomes between the four and five trocars were compared to evaluate the impact of trocar layout on the surgical outcomes. Results:There were 91(95.8%) cases successfully undergoing P. R.E.S.S. BCE technique, with one case converted to open BCE due to bleeding and three cases converted to distal ureter Hem-o-lok clipping because of poor exposure. Median operative time was 180(125, 230)min, and estimated blood loss was 100(50, 100)ml. The overall complication rate was 10.5%(10/95), including 2 cases(2.1%) of intraoperative bleeding, with 1 case treated by transfusion (400 ml), the other case converted to open surgery without transfusion. There were 8 cases of postoperative complications(8.4%), including 7 cases of Clavien-Dindo grade Ⅱ(3 cases of secondary hemorrhage, one case for each of drug allergy, acute renal insufficiency, blood creatinine increased to 490 μmol/L, or lung infection with lymphatic leakage), 1 case of grade Ⅲa(intestinal obstruction treated with insertion of the intestinal obstruction catheter under local anesthesia), and all these patients were discharged smoothly. The difference between the four and five trocars was not statistically significant in the following variables, including the rate of surgical conversion(8.3% vs. 1.7%), estimated intraoperative blood loss(100 ml vs. 60 ml), ratio of intraoperative lymph node dissection (25.0% vs.20.3%), P. R.E.S.S. bladder cuff excision success rate(91.7% vs.98.3%), the incidence of intraoperative and postoperative complications (13.8% vs.8.5%), pT 1-2/pT 3-4/pTis stage(22/11/3 cases vs.37/19/3 cases) and the proportion of recurrence or metastasis(8.3% vs.3.4%)(all P>0.05). However, the differences in the operation time(190 min vs.170 min, P=0.011)and postoperative hospital stay(5 d vs.6 d, P=0.005) were statistically significant. Conclusions:P. R.E.S.S. bladder cuff resection technique is safe and feasible during the procedure of pure retroperitoneoscopic radical nephroureterectomy by a single surgical position and facilitates seeable adequate bladder cuff excision by establishing an enlarged pelvic lateral extraperitoneal space and pneumovesicum. Five-trocar technique is more suitable for patients with lower ureteral tumors but may be associated with a longer postoperative hospital stay compared with the four-trocar technique.

Methylotrophic yeasts are considered as promising cell factories for bio-manufacturing due to their several advantages such as tolerance to low pH and high temperature. In particular, their methanol utilization ability may help to establish a methanol biotransformation process, which will expand the substrate resource for bio-refinery and the product portfolio from methanol. This review summarize current progress on engineering methylotrophic yeasts for production of proteins and chemicals, and compare the strengths and weaknesses with the model yeast Saccharomyces cerevisiae. The challenges and possible solutions in metabolic engineering of methylotrophic yeasts are also discussed. With the developing efficient genetic tools and systems biology, the methylotrophic yeasts should play more important roles in future green bio-manufacturing.
Metabolic Engineering ; Methanol ; Saccharomyces cerevisiae/genetics* ; Yeasts

Objective To analyze the risk factors of renal replacement therapy (RRT) in acute kidney injury (AKI) patients after liver transplantation, and to investigate the prognosis effect of initial RRT treatment time. Methods Clinical data of 132 recipients undergoing organ donation for cardiac death (DCD) allograft orthotopic liver transplantation admitted to Ningbo Medical Center Lihuili Hospital and Ningbo Medical Center Lihuili Eastern Hospital from July 2014 to July 2018 was retrospectively analyzed. AKI was defined and staged by the criteria of Kidney Disease Improving Global Outcomes (KDIGO) guideline in the first 7 days. According to the implementation of RRT, the patients were divided into non-RRT group and RRT group. The differences in gender, age, body mass index (BMI), model for end-stage liver disease with serum sodium (MELD-Na) score, serum creatinine (SCr), and intraoperative norepinephrine (NE) dose, blood loss, fluid infusion, anhepatic phase time, duration of operation between two groups were compared. The statistically significant risk factors of AKI found by univariate analysis were selected and analyzed to find independent risk factors of RRT in AKI patients after liver transplantation with multivariate Logistic regression analysis. The receiver operating characteristic (ROC) curve was drawn to evaluate the test efficiency of all risk factors of RRT implementation. According to the implementation of RRT on KDIGO stage-2, all the patients on KDIGO stage-2 and stage-3 were divided into early group (initial RRT on KDIGO stage-2) and delayed group (including self-improvement without RRT on KDIGO stage-2 and initial RRT on KDIGO stage-3). The duration of mechanical ventilation, the length of intensive care unit (ICU) stay, AKI duration, incidence of catheter related bloodstream infection (CRBSI) and 28-day mortality were compared between the two groups. Results All 132 receptors were enrolled in the final analysis, and 77 patients developed AKI, accounting for 58.3%, among which 52 cases were in RRT group (67.5%) and 25 were in non-RRT group (32.5%). As shown by univariate analysis, the MELD-Na score (21.6±4.4 vs. 18.0±4.3), intraoperative NE dose (μg·kg-1·h-1: 7.5±1.2 vs. 5.2±1.7), blood loss [mL: 3 000 (2 200, 4 000) vs. 2 600 (1 800, 3 200)], fluid infusion [mL: 6 400 (4 500, 7 800) vs. 5 600 (4 200, 6 800)], and anhepatic period (minutes: 65.6±4.5 vs. 63.0±5.0) were significantly increased in RRT group as compared with those in non-RRT group (all P < 0.05). There was no significant difference in gender, age, BMI, SCr before operation or the duration of operation. It was shown by multivariate Logistic regression analysis that MELD-Na score before operation [odds ratio (OR) = 1.398, 95% confidence interval (95%CI) = 1.062-1.841, P = 0.017], intraoperative NE dose (OR = 4.724, 95%CI = 2.036-10.961, P = 0.000) and fluid infusion (OR = 1.002, 95%CI = 1.001-1.004, P = 0.010) were independent risk factors of RRT implementation in AKI patients after liver transplantation. It was shown by ROC curve analysis that the area under the ROC curve (AUC) of MELD-Na score, NE dose and fluid infusion for predicting the implementation of RRT in AKI patients after liver transplantation was 0.719, 0.867, and 0.670, respectively, which suggesting that NE dose had moderate predictive value, but MELD-Na score and fluid infusion had low predicative value. When the optimal cut-off value of NE dose was 6.5 μg·kg-1·h-1, the sensitivity was 84.6% and the specificity was 80.0%. The 28-day mortality was both 0 in early group (n = 25) and delayed group (n = 39). Compared with the early group, the duration of mechanical ventilation (hours: 41.0±1.0 vs. 35.8±6.7) and the length of ICU stay (hours: 98.8±6.6 vs. 94.2±7.3) were significantly increased in delayed group (both P < 0.05), there was no significant difference in AKI duration (days: 11.8±4.2 vs. 10.6±4.9) or the incidence of CRBSI [5.1% (2/39) vs. 4.0% (1/25), both P > 0.05]. Conclusions MELD-Na score, intraoperative NE dose and fluid infusion were the independent risk factors of RRT implementation in AKI patients after liver transplantation. NE dose had moderate predictive value, but MELD-Na score and fluid infusion had low predicative value. Initial RRT on KDIGO stage-2 could reduce the duration of mechanical ventilation and the length of ICU stay.

Objective To study the short-term effectiveness of robotic-assisted laparoscopic radical prostatectomy in high-risk prostate cancer.Methods From March 2012 to March 2017,400 patients with high-risk prostate who underwent robotic-assisted laparoscopic radical prostatectomy were reviewed.The median age was 68 years old(ranged from 49 to 83 years),and the median PSA was 23.1 ng/ ml(ranged from 5.2 to 999.0 ng/ml).Preoperative parameters,surgical interventional data,postoperative pathology and follow-up data were collected.Logistic regression was used to analyze the risk factors of positive surgical margin in postoperative pathology.Results All the operations were successfully completed.Median operation time was 115 min(ranged 50-555 min),and median estimated blood loss was 110 ml(ranged 30-500 ml).Six patients had perioperative complications,among which two were rectal injury,two were cardio-cerebrovascular disease and two were hemorrhage.There was no perioperative death.Positive surgical margin was detected in 151 patients,accounting for 37.8％.A total of 345 cases (86.3％) underwent lymphadenectomy,of which 253 cases (63.3％) were performed standardized resection and 92 cases (23％) were performed extensive resection.The median number of resected lymph nodes was 9 (ranged 3-36).Eighty cases (23.2％,80/345) were positive in resected lymph nodes.Regression analysis showed that preoperative PSA ＞ 20 ng / ml or clinical stage ≥ T2c were risk factors for positive surgical margins.After a median follow-up of 14.4 months (ranged 2.0-58.8 months),the overall incidence of biochemical recurrence was 33.4％ (107/320),and the urinary continence rate one year after operation was 86.6％ (277/320).Conclusions Robotic-assisted laparoscopic radical prostatectomy in patients with high-risk prostate cancer was a feasible,safe and effective approach.Preoperative PSA and clinical stage were the risk factors for positive surgical margin.

Objective: To investigate the effect of sevolfurane (SEVO) post-conditioning on protein kinase B (AKT)/mammalian target of rapamycin (mTOR) pathway for protecting ischemia/reperfusion (I/R) injury in isolated rat’s heart. Methods: A total of 84 isolated rat’s heart prepared by Langendorff method were randomly divided into 7 groups andn=12 in each group.①Sham group,②I/R group,③SEVO post- conditioning (SPC) group,④NVP-BEZ235 solvent dimenthyl sulfoxide (DMSO) group,⑤Phosphatidyl inositol 3-kinase (PI3K)/mTOR dual inhibitor NVP-BEZ235 (BEZ) group,⑥BEZ+SPC group and⑦SEVO alone group. The hearts received 30 min ischemia followed by 120 min reperfusion with relevant treatment except for Sham group and SEVO group in which the hearts were without ischemia process. Myocardial infarct (MI) size and tissue pathological changes were observed, protein expressions of phosphor-AKT (P-AKT)/total-AKT, P-mTOR/total-mTOR, Beclin1, Bax/Bcl-2 and cleaved Caspase-3 were examined at the end of reperfusion respectively. Results: Compared with I/R group, SPC group presented decreased MI size (26.28±4.00) % vs (49.22±3.66) % and reduced tissue pathological changes; up-regulated protein expressions of P-AKT/total-AKT and P-mTOR/total-mTOR by 79.85% and 67.02%, while down-regulated protein expressions of Beclin1, Bax/Bcl-2 and cleaved Caspase-3 by 33.77%, 69.26% and 48.84%respectively, allP<0.05. Compared with SPC group, BEZ+SPC group sowed increased MI size (53.85±4.06) % vs (26.28±4.00) %and elevated tissue pathological changes; down-regulated proteins expressions of P-AKT/total-AKT and P-mTOR/total-mTOR by 46.06% and 42.95%, while up-regulated protein expressions of Beclin1, Bax/Bcl-2 and cleaved Caspase-3 by 29.90%, 206.85% and 114.65% respectively, allP<0.05. Conclusion: SPC may activate AKT/mTOR pathway and inhibit cardiomyocyte autophagy and apoptosis, thereby attenuate I/R injury in isolated rats’ heart.

Objective To investigate the role of AKT/GSK3β/mTOR signaling molecule in myocardial protection of sevoflurane postconditioning.Methods Thirty-nine male Sprague-Dawley rats,weighing 200-250 g,installed in vivo myocardial ischemia-reperfusion model by left anterior de-scending coronary occlusion for 30 min.Rat hearts were randomly divided into 3 groups (n = 13 ):sham control group (group Sham),purely ischemia-reperfusion group (group IR),sevoflurane post-conditioning group (group SPC).With the exception of group Sham,each group was subjected to oc-clusion for 30 min followed by 2 h reperfusion.Group SPC was subjected to sevoflurane postcondi-tioning:2.4% sevoflurane was inhaled for 1 5 min starting from the end of ischemia until 1 5 min after beginning of reperfusion,while 33% oxygen was inhaled in the other groups.At the end of 2 h reper-fusion,cardiac function was evaluated by two-dimensional echocardiography,myocardial infarction size was measured by using 1% 2,3,5-triphenyltetrazolium chloride triazole (TTC),myocardial ultra-structural alterations was detected by transmission electron microscopy (TEM),cardiomyocytes ap-optosis was examined by terminal deoxynucleotidyl nickend labeling (TUNEL),the expressions of p-AKT/t-AKT, p-GSK3β/t-GSK3β, p-mTOR/t-mTOR,Bcl-2 and Bax proteins was measured by Western blot.Results Compared with group Sham,cardiac function was deteriorated,myocardial in-farct size was increased,cardiomyocyte mitochondrial damage was increased,positive apoptotic car-diomyocyte was increased,the expression of Bcl-2 was down-regluated,and the expressions of p-AKT/t-AKT,p-GSK3β/t-GSK3β,p-mTOR/t-mTOR and Bax were up-regluated in group IR (P <0.05).Compared with group IR,cardiac function was improved,myocardial infarct size was de-creased,cardiomyocyte mitochondrial damage was decreased,positive apoptotic cardiomyocyte was decreased,the expression of Bax was down-regluated,and the expressions of p-AKT/t-AKT,p-GSK3β/t-GSK3β,p-mTOR/t-mTOR and Bcl-2 were up-regluated in group SPC (P < 0.05 ). Conclusion Sevoflurane postconditioning can mitigate ischemia-reperfusion injury to in vivo rat hearts,decreased cardiomyocyte mitochondrial damage,inhibited cardiomyocyte apoptosis,and its mechanism was related to the activation of AKT/GSK3β/mTOR signaling molecule.