One of the technical bottlenecks limiting the high yield of 1,4-butanediamine is the insufficient tolerance of strains to 1,4-butanediamine. Enhancing the tolerance of strains to 1,4-butanediamine is therefore a primary challenge that needs to be addressed for the construction of strains with high yields of 1,4-butanediamine. Staphylococcus pasteuri 326180 exhibits exceptional tolerance to high-concentration 1,4-butanediamine, serving as both an ideal model for studying the mechanism underlying the 1,4-butanediamine tolerance and a novel host for constructing strains capable of efficiently producing 1,4-butanediamine. However, for both the research on the tolerance mechanism and the modification of chassis strains, gene editing of S. pasteuri needs to be carried out at the molecular level. The research objective of this paper is to establish a genetic manipulation system for S. pasteuri, laying foundation for subsequent studies on tolerance mechanism and the modification of chassis strains. This study systematically optimized the electroporation conditions, including key parameters such as the growth phase of cells, electric field strength, electroporation buffer, and recovery medium, successfully establishing an electroporation method for S. pasteuri. Additionally, we constructed the gene editing plasmid pCpfOA by replacing the resistance expression cassette, optimized the selection markers for gene editing, and finally established a CRISPR/Cpf1-based gene editing technology for S. pasteuri, achieving an editing efficiency of 90%. The genetic manipulation system of S. pasteuri established in this study provides technical support for research into the tolerance mechanism of this bacterium and the genetic modification of chassis strains.
Staphylococcus/drug effects* ; Gene Editing/methods* ; Electroporation/methods* ; Plasmids/genetics* ; CRISPR-Cas Systems ; Genetic Engineering/methods*

Objective:Based on the activities of Helicobacter pylori ( HP) screening in Jiangsu Province in 2024, to evaluate the overall management ability in HP screening, testing, treatment and follow-up in primary medical facilities. Methods:From May 15 to October 18, 2024, the data of HP screening and treatment were retrospectively collected from 79 township health centers, community hospitals, and community health service centers in Jiangsu Province. The rates of screening completion, urea breath test (UBT) completion, treatment rate, UBT follow-up completion, and HP eradication were analyzed. Chi-square test was used for statistical analysis. Results:The completion rate of HP screening was 94.45% (15 489/16 400). There were 6 604 cases (42.64%) with serum HP antibody positive among the 15 489 individuals who completed screening. The positive rate of serum HP antibody in males was higher than that in females (44.77%, 2 643/5 904 vs. 41.32%, 3 961/9 585), and the difference was statistically significant ( χ2=17.69, P<0.001). The positive rates of serum HP antibody in screened individuals aged 18 to 19, 20 to 39, 40 to 59, and 60 to 75 years old were 22.38% (32/143), 36.12% (1 168/3 234), 45.01% (3 240/7 199), and 44.05% (2 164/4 913), respectively, and the difference was statistically significant( χ2=100.73, P<0.001). Among the 6 604 HP antibody-positive individuals, 4 381 cases completed UBT, with a UBT completion rate of 66.34% (4 381/6 604). There were 3 197 individuals with both HP serum antibody and UBT positive, the consistency rate of the 2 tests was 72.97% (3 197/4 381). Totally 2 737 cases received treatment, with a treatment completion rate of 85.61% (2 737/3 197); 2 327 individuals underwent UBT follow-up, with a follow-up completion rate of 85.02% (2 327/2 737). During follow-up, the result of UBT was negative in 1 982 individuals, and the HP eradication rate was 85.17% (1 982/2 327). Conclusions:There are deficiencies in the completion rate of HP screening, testing, treatment, and follow-up in primary hospitals, especially in the completion rate of UBT, which may be related to cognitive insufficiency for HP in residents. It is necessary to strengthen the training of physicians′ abilities in primary hospitals, optimize the allocation of drug resources, enhance health education, and increase residents′ participation and compliance.

Objective:To investigate the clinical application effect and preliminary experience of ureteroscopy-assisted flexible ureteroscopic lithotripsy in the treatment of transplanted kidney stones.Method:A retrospective analysis was conducted on the clinical data of 9 kidney transplant recipients with graft stones who underwent ureteroscopy-assisted flexible ureteroscopic lithotripsy at the First Affiliated Hospital of Zhengzhou University between January 2020 and January 2023. The recipients' general information, surgical procedures, and postoperative outcomes were reviewed and summarized. Additionally, a comprehensive literature search was performed in both Chinese and international databases, including CNKI, Wanfang, and the China Academic Journals Full-text Database, as well as PubMed and Web of Science. The search terms included " kidney transplant" "flexible ureteroscope" "ureteroscope" and " urinary tract stones" . The search covered all publications available up to December 2023.Result:The 9 recipients had a mean age of 37. 56 years (range: 27–54 years) . The onset time of the kidney stones ranged from 1 to 13 months after kidney transplantation. All recipients were diagnosed with transplanted kidney stones by CT and showed no symptoms of kidney colic at the time of onset. All transplanted kidneys were located in the right iliac fossa, with varying degrees of hydronephrosis. The smallest stone diameter was 0. 5 cm, and the largest was 1. 3 cm. The number of stones ranged from 1 to 3 per recipient; 6 cases had solitary stones, and 3 had multiple stones in the renal pelvis and calyces. In all 9 surgeries, the ureteral orifice of the transplanted kidney was located at the apex of the bladder using ureteroscopy, and a hydrophilic guidewire was inserted into the renal pelvis. A second ureteroscopy was then performed, guiding the ureteroscope tip into the transplanted ureteral orifice in coaxial alignment with the guidewire. A flexible ureteroscope sheath was advanced along the guidewire, followed by the insertion of a digital flexible ureteroscope to perform holmium laser dusting lithotripsy. Stone composition analysis revealed mixed calcium oxalate monohydrate and calcium oxalate dihydrate. The surgical duration ranged from 30 to 75 minutes. No complications such as massive bleeding, septic shock, or rejection occurred. Postoperative review at 1 month, after ureteral stent removal, showed no residual stones in any patient. During 6 months of follow-up, no recurrence of stones was observed, and the renal function of the transplanted kidneys remained stable compared to preoperative levels. A total of 19 related articles were identified through literature review, including 11 in English and 8 in Chinese. Chinese reports on transplanted kidney stones were all from single-center studies with small case numbers. In contrast, foreign meta-analyses and multicenter studies showed that treatments for transplanted kidney stones abroad mainly included antegrade/retrograde ureteroscopy and percutaneous nephroscopy.Conclusion:Ureteroscopy-assisted flexible ureteroscopic lithotripsy is a feasible treatment for transplanted kidney stones. It can effectively protect graft kidney function and has promising clinical application prospects.

Objective:Based on the activities of Helicobacter pylori ( HP) screening in Jiangsu Province in 2024, to evaluate the overall management ability in HP screening, testing, treatment and follow-up in primary medical facilities. Methods:From May 15 to October 18, 2024, the data of HP screening and treatment were retrospectively collected from 79 township health centers, community hospitals, and community health service centers in Jiangsu Province. The rates of screening completion, urea breath test (UBT) completion, treatment rate, UBT follow-up completion, and HP eradication were analyzed. Chi-square test was used for statistical analysis. Results:The completion rate of HP screening was 94.45% (15 489/16 400). There were 6 604 cases (42.64%) with serum HP antibody positive among the 15 489 individuals who completed screening. The positive rate of serum HP antibody in males was higher than that in females (44.77%, 2 643/5 904 vs. 41.32%, 3 961/9 585), and the difference was statistically significant ( χ2=17.69, P<0.001). The positive rates of serum HP antibody in screened individuals aged 18 to 19, 20 to 39, 40 to 59, and 60 to 75 years old were 22.38% (32/143), 36.12% (1 168/3 234), 45.01% (3 240/7 199), and 44.05% (2 164/4 913), respectively, and the difference was statistically significant( χ2=100.73, P<0.001). Among the 6 604 HP antibody-positive individuals, 4 381 cases completed UBT, with a UBT completion rate of 66.34% (4 381/6 604). There were 3 197 individuals with both HP serum antibody and UBT positive, the consistency rate of the 2 tests was 72.97% (3 197/4 381). Totally 2 737 cases received treatment, with a treatment completion rate of 85.61% (2 737/3 197); 2 327 individuals underwent UBT follow-up, with a follow-up completion rate of 85.02% (2 327/2 737). During follow-up, the result of UBT was negative in 1 982 individuals, and the HP eradication rate was 85.17% (1 982/2 327). Conclusions:There are deficiencies in the completion rate of HP screening, testing, treatment, and follow-up in primary hospitals, especially in the completion rate of UBT, which may be related to cognitive insufficiency for HP in residents. It is necessary to strengthen the training of physicians′ abilities in primary hospitals, optimize the allocation of drug resources, enhance health education, and increase residents′ participation and compliance.

Objective:To investigate the clinical application effect and preliminary experience of ureteroscopy-assisted flexible ureteroscopic lithotripsy in the treatment of transplanted kidney stones.Method:A retrospective analysis was conducted on the clinical data of 9 kidney transplant recipients with graft stones who underwent ureteroscopy-assisted flexible ureteroscopic lithotripsy at the First Affiliated Hospital of Zhengzhou University between January 2020 and January 2023. The recipients' general information, surgical procedures, and postoperative outcomes were reviewed and summarized. Additionally, a comprehensive literature search was performed in both Chinese and international databases, including CNKI, Wanfang, and the China Academic Journals Full-text Database, as well as PubMed and Web of Science. The search terms included " kidney transplant" "flexible ureteroscope" "ureteroscope" and " urinary tract stones" . The search covered all publications available up to December 2023.Result:The 9 recipients had a mean age of 37. 56 years (range: 27–54 years) . The onset time of the kidney stones ranged from 1 to 13 months after kidney transplantation. All recipients were diagnosed with transplanted kidney stones by CT and showed no symptoms of kidney colic at the time of onset. All transplanted kidneys were located in the right iliac fossa, with varying degrees of hydronephrosis. The smallest stone diameter was 0. 5 cm, and the largest was 1. 3 cm. The number of stones ranged from 1 to 3 per recipient; 6 cases had solitary stones, and 3 had multiple stones in the renal pelvis and calyces. In all 9 surgeries, the ureteral orifice of the transplanted kidney was located at the apex of the bladder using ureteroscopy, and a hydrophilic guidewire was inserted into the renal pelvis. A second ureteroscopy was then performed, guiding the ureteroscope tip into the transplanted ureteral orifice in coaxial alignment with the guidewire. A flexible ureteroscope sheath was advanced along the guidewire, followed by the insertion of a digital flexible ureteroscope to perform holmium laser dusting lithotripsy. Stone composition analysis revealed mixed calcium oxalate monohydrate and calcium oxalate dihydrate. The surgical duration ranged from 30 to 75 minutes. No complications such as massive bleeding, septic shock, or rejection occurred. Postoperative review at 1 month, after ureteral stent removal, showed no residual stones in any patient. During 6 months of follow-up, no recurrence of stones was observed, and the renal function of the transplanted kidneys remained stable compared to preoperative levels. A total of 19 related articles were identified through literature review, including 11 in English and 8 in Chinese. Chinese reports on transplanted kidney stones were all from single-center studies with small case numbers. In contrast, foreign meta-analyses and multicenter studies showed that treatments for transplanted kidney stones abroad mainly included antegrade/retrograde ureteroscopy and percutaneous nephroscopy.Conclusion:Ureteroscopy-assisted flexible ureteroscopic lithotripsy is a feasible treatment for transplanted kidney stones. It can effectively protect graft kidney function and has promising clinical application prospects.

Objective: QL1604 is a highly selective, humanized monoclonal antibody against programmed death protein 1. We assessed the efficacy and safety of QL1604 plus chemotherapy as first-line treatment in patients with advanced cervical cancer. Methods: This was a multicenter, open-label, single-arm, phase II study. Patients with advanced cervical cancer and not previously treated with systemic chemotherapy were enrolled to receive QL1604 plus paclitaxel and cisplatin/carboplatin on day 1 of each 21-day cycle for up to 6 cycles, followed by QL1604 maintenance treatment. Results: Forty-six patients were enrolled and the median follow-up duration was 16.5 months. An 84.8% of patients had recurrent disease and 13.0% had stage IVB disease. The objective response rate (ORR) per Response Evaluation Criteria in Advanced Solid Tumors (RECIST) v1.1 was 58.7% (27/46). The immune ORR per immune RECIST was 60.9% (28/46).The median duration of response was 9.6 months (95% confidence interval [CI]=5.5–not estimable). The median progression-free survival was 8.1 months (95% CI=5.7–14.0). Fortyfive (97.8%) patients experienced treatment-related adverse events (TRAEs). The most common grade≥3 TRAEs (>30%) were neutrophil count decrease (50.0%), anemia (32.6%), and white blood cell count decrease (30.4%). Conclusion QL1604 plus paclitaxel-cisplatin/carboplatin showed promising antitumor activity and manageable safety profile as first-line treatment in patients with advanced cervical cancer. Programmed cell death protein 1 inhibitor plus chemotherapy may be a potential treatment option for the patient population who have contraindications or can’t tolerate bevacizumab, which needs to be further verified in phase III confirmatory study.

Objective: QL1604 is a highly selective, humanized monoclonal antibody against programmed death protein 1. We assessed the efficacy and safety of QL1604 plus chemotherapy as first-line treatment in patients with advanced cervical cancer. Methods: This was a multicenter, open-label, single-arm, phase II study. Patients with advanced cervical cancer and not previously treated with systemic chemotherapy were enrolled to receive QL1604 plus paclitaxel and cisplatin/carboplatin on day 1 of each 21-day cycle for up to 6 cycles, followed by QL1604 maintenance treatment. Results: Forty-six patients were enrolled and the median follow-up duration was 16.5 months. An 84.8% of patients had recurrent disease and 13.0% had stage IVB disease. The objective response rate (ORR) per Response Evaluation Criteria in Advanced Solid Tumors (RECIST) v1.1 was 58.7% (27/46). The immune ORR per immune RECIST was 60.9% (28/46).The median duration of response was 9.6 months (95% confidence interval [CI]=5.5–not estimable). The median progression-free survival was 8.1 months (95% CI=5.7–14.0). Fortyfive (97.8%) patients experienced treatment-related adverse events (TRAEs). The most common grade≥3 TRAEs (>30%) were neutrophil count decrease (50.0%), anemia (32.6%), and white blood cell count decrease (30.4%). Conclusion QL1604 plus paclitaxel-cisplatin/carboplatin showed promising antitumor activity and manageable safety profile as first-line treatment in patients with advanced cervical cancer. Programmed cell death protein 1 inhibitor plus chemotherapy may be a potential treatment option for the patient population who have contraindications or can’t tolerate bevacizumab, which needs to be further verified in phase III confirmatory study.

Objective: QL1604 is a highly selective, humanized monoclonal antibody against programmed death protein 1. We assessed the efficacy and safety of QL1604 plus chemotherapy as first-line treatment in patients with advanced cervical cancer. Methods: This was a multicenter, open-label, single-arm, phase II study. Patients with advanced cervical cancer and not previously treated with systemic chemotherapy were enrolled to receive QL1604 plus paclitaxel and cisplatin/carboplatin on day 1 of each 21-day cycle for up to 6 cycles, followed by QL1604 maintenance treatment. Results: Forty-six patients were enrolled and the median follow-up duration was 16.5 months. An 84.8% of patients had recurrent disease and 13.0% had stage IVB disease. The objective response rate (ORR) per Response Evaluation Criteria in Advanced Solid Tumors (RECIST) v1.1 was 58.7% (27/46). The immune ORR per immune RECIST was 60.9% (28/46).The median duration of response was 9.6 months (95% confidence interval [CI]=5.5–not estimable). The median progression-free survival was 8.1 months (95% CI=5.7–14.0). Fortyfive (97.8%) patients experienced treatment-related adverse events (TRAEs). The most common grade≥3 TRAEs (>30%) were neutrophil count decrease (50.0%), anemia (32.6%), and white blood cell count decrease (30.4%). Conclusion QL1604 plus paclitaxel-cisplatin/carboplatin showed promising antitumor activity and manageable safety profile as first-line treatment in patients with advanced cervical cancer. Programmed cell death protein 1 inhibitor plus chemotherapy may be a potential treatment option for the patient population who have contraindications or can’t tolerate bevacizumab, which needs to be further verified in phase III confirmatory study.

BACKGROUND:Acellular vascular scaffolds can mimic the microstructure and function of native blood vessels,but some extracellular matrix loss occurs during their preparation,which affects their hemocompatibility.Therefore,it is necessary to modify them to improve their hemocompatibility. OBJECTIVE:To assess the hemocompatibility of acellular vascular scaffold prepared by Triton-x100/heparin sodium treatment. METHODS:The abdominal aorta was taken from SD rats and randomly divided into control and experimental groups.The control group was treated with Triton-x100 for 48 hours.The experimental group was treated with Triton-x100 for 48 hours and then treated with heparin sodium.The morphology and hydrophilicity of the two groups of acellular vascular scaffolds were detected.The hemocompatibility of the two groups of acellular vascular scaffold was evaluated by recalcification coagulation time test,platelet adhesion test,dynamic coagulation time test,hemolysis test,and complement activation test. RESULTS AND CONCLUSION:(1)Scanning electron microscopy showed that the surface of the two groups of vascular scaffolds was relatively intact,and a large number of fiber filaments appeared on the surface of the scaffolds after decellularity treatment,and the surface microstructure changed significantly.The water contact angle of the two groups of vascular scaffolds was smaller than that of natural vessels(P<0.000 1).There was no significant difference in water contact angle between the two groups(P>0.05).(2)The coagulation time of vascular scaffold was longer in the experimental group than in the control group(P<0.05).The number of platelets attached to the scaffold membrane was less in the experimental group than that in the control group(P<0.000 1).The coagulation index was greater in the experimental group than that in the control group(P<0.01),and the complement level was lower in the experimental group than that in the control group(P<0.001).The hemolysis rate of the two groups was lower than 5%of the national standard.(3)To conclude,acellular scaffold treated with Triton-x100/heparin sodium has excellent hemocompatibility.

Objective:To analyze the influencing factors of genotypic drug resistance mutations in people living with human immunodeficiency virus and acquired immunodeficiency syndrome(PLWHA) who failed anti-retroviral therapy (ART) in Henan Province, in order to provide a basis for adjusting ART regimens and reducing drug resistance.Methods:PLWHA with virological failure (human immunodeficiency virus (HIV) RNA≥500 copies/mL) after receiving ART for more than 24 weeks were included in Henan Province from January 2018 to December 2022. Baseline CD4 + T lymphocyte counts, ART regimens and other clinical data were collected. HIV-1 gene subtypes and their drug resistance sequence mutations were detected in the Sixth People′s Hospital of Zhengzhou, and the sequences were submitted to the HIV Drug Resistance Interpretation System of Stanford University for comparison of test results. Genotypic drug resistance to nucleotide reverse transcriptase inhibitors (NRTI), non-nucleoside reverse transcriptase inhibitors (NNRTI), protease inhibitors (PI) and integrase inhibitors (INSTI) was determined. Multivariate logistic regression was used to analyze the influencing factors of drug resistance in patients with ART failure. Results:Among 982 PLWHA, the sequences of 899 cases were successfully amplified, and drug resistance was detected in 737 cases, with the drug resistance rate of 81.98%(737/899). The rates of resistance to NRTIs, NNRTIs, PIs and INSTIs were 71.97%(647/899), 79.31%(713/899), 5.23%(47/899) and 2.72%(20/734), respectively.The largest number of those who developed concomitant resistance to two classes of drugs was 588 cases (79.78%), mainly NRTI and NNRTI concomitant resistance in 583 cases (79.10%). There were 99 cases (13.43%) who developed resistance to only one class of drugs, and those who developed concurrent resistance to three classes of drugs were 48 cases (6.51%), and two cases (0.27%) were found to be resistant to all four classes of drugs mentioned above. A total of 10 HIV genotypes were detected, among which subtype B accounted for the most (59.73%(537/899)), followed by circulating recombinant form (CRF)01_AE subtype (21.91%(197/899)) and CRF07_BC subtype (9.45%(85/899)). The risk factors affecting the development of drug resistance were baseline CD4 + T lymphocyte counts, ART regimens and HIV-1 genotypes. The risk of drug resistance in patients with baseline CD4 + T lymphocyte counts <100/μL was 4.55 times (95% confidence interval ( CI) 2.69 to 7.70) higher than patients with CD4 + T lymphocyte counts≥250/μL, the risk of drug resistance in patients using 2NRTIs+ NNRTI regimen was 4.51 times (95% CI 1.75 to 11.63) higer than those using 2NRTIs+ INSTI regimen, and patients infected with B and CRF01_AE subtype was 2.18 times (95% CI 1.10 to 4.29) and 2.70 times (95% CI 1.26 to 5.78) higer than those with CRF07_BC subtype, respectively. Conclusions:The incidence of genotypic drug resistance in PLWHA with ART failure in Henan Province is high. Low baseline CD4 + T lymphocyte counts, 2NRTIs+ NNRTI regimens, and genotype B and CRF01_AE are risk factors for drug resistance in PLWHA.