BACKGROUND:Solute carrier family 1 member 5(SLC1A5)plays a potential role in a variety of diseases,but the exact mechanism of action is unclear.The construction of stable SLC1A5 overexpression and knockdown cell models can provide a powerful experimental tool for in-depth study of the exact role and mechanism of SLC1A5 in diseases and the discovery of potential therapeutic targets. OBJECTIVE:To construct lentiviral vectors for overexpression and knockdown of mouse SLC1A5 and establish stable transfected RAW264.7 cell lines,so as to provide an experimental foundation for further investigation of the role of SLC1A5 in inflammation. METHODS:Primers were designed and synthesized based on the SLC1A5 gene sequence,and the gene segment was amplified using polymerase chain reaction.Subsequently,the target gene segment was directionally inserted into the GV492 vector plasmid,which had been digested with AgeI/NheI enzymes,to construct recombinant lentiviral plasmids.Positive clones were further selected,and their sequences were confirmed.The pHelper1.0 plasmid vector and pHelper2.0 plasmid vector,along with the target plasmid vector,was co-cultured with 293T cells for transfection,resulting in the production and titration of lentiviral stocks.Furthermore,RAW264.7 cells were cultured in vitro,and the working concentration of puromycin was determined.Lentiviruses were separately co-cultured with RAW264.7 cells,and transfection efficiency was determined by measuring fluorescence intensity.Stable transfected cells were selected using puromycin,and real-time fluorescence quantitative PCR and western blot assay were employed to assess the gene and protein expression levels of SLC1A5 in stably transfected cell lines. RESULTS AND CONCLUSION:(1)Sequencing results indicated a perfect match between the sequencing and target sequences,confirming the successful construction of recombinant lentiviral vectors.(2)The titer for the overexpression SLC1A5 lentivirus was 1×109 TU/mL,while the titer for the knockdown SLC1A5 lentivirus was 3×109 TU/mL.(3)The working concentration of puromycin for RAW264.7 cells was determined to be 3 μg/mL.(4)The optimal conditions for transfecting RAW264.7 cells with overexpression/knockdown expression of SLC1A5 lentivirus involved the use of HiTransG P transfection enhancer with a multiplicity of infection value of 50.(5)A significant upregulation of the gene and protein expression levels of SLC1A5 was detected in cell lines stably overexpressing SLC1A5,while gene and protein expression levels of SLC1A5 were significantly decreased in the knockdown stable cell lines.These findings indicate that lentiviral vectors for mouse SLC1A5 overexpression and knockdown have been successfully constructed and a stably transfected RAW264.7 cell line has been obtained.

Methods: This retrospective study analyzed 32 consecutive IDS patients who underwent UEDD surgery. Clinical features, laboratory data (erythrocyte sedimentation rate and C-reactive protein), and treatment outcomes were analyzed. Results: Definite microorganisms were identified in 27 patients (84.3%), with 24 (88.9%) meeting cure criteria. The cure rate was significantly higher in the detected pathogen group compared to the undetected pathogen group (88.9% vs. 80%; χ²=19.36, p<0.0001). Metagenomic next generation sequencing (mNGS) provided faster diagnosis (41.72±6.81 hours) compared to tissue culture (95.74±35.47 hours, p<0.05). The predominant causative pathogen was Mycobacterium tuberculosis, followed by Staphylococcus aureus. Significant improvements were observed in Visual Analog Scale pain scores, from a mean of 7.9 preoperatively to 1.06 at 1 year postoperatively. The Oswestry Disability Index revealed a similar trend, showing significant improvement (p<0.05). Conclusions UEDD is a viable alternative to traditional open surgery for managing IDS in high-risk patients. UEDD offers a dual therapeutic-diagnostic advantage during the initial admission phase, enabling simultaneous debridement, neurological decompression, and targeted biopsy in a single intervention. Compared with traditional tissue culture, mNGS enables rapid microbiological diagnosis and extensive pathogen coverage.

Methods: This retrospective study analyzed 32 consecutive IDS patients who underwent UEDD surgery. Clinical features, laboratory data (erythrocyte sedimentation rate and C-reactive protein), and treatment outcomes were analyzed. Results: Definite microorganisms were identified in 27 patients (84.3%), with 24 (88.9%) meeting cure criteria. The cure rate was significantly higher in the detected pathogen group compared to the undetected pathogen group (88.9% vs. 80%; χ²=19.36, p<0.0001). Metagenomic next generation sequencing (mNGS) provided faster diagnosis (41.72±6.81 hours) compared to tissue culture (95.74±35.47 hours, p<0.05). The predominant causative pathogen was Mycobacterium tuberculosis, followed by Staphylococcus aureus. Significant improvements were observed in Visual Analog Scale pain scores, from a mean of 7.9 preoperatively to 1.06 at 1 year postoperatively. The Oswestry Disability Index revealed a similar trend, showing significant improvement (p<0.05). Conclusions UEDD is a viable alternative to traditional open surgery for managing IDS in high-risk patients. UEDD offers a dual therapeutic-diagnostic advantage during the initial admission phase, enabling simultaneous debridement, neurological decompression, and targeted biopsy in a single intervention. Compared with traditional tissue culture, mNGS enables rapid microbiological diagnosis and extensive pathogen coverage.

Methods: This retrospective study analyzed 32 consecutive IDS patients who underwent UEDD surgery. Clinical features, laboratory data (erythrocyte sedimentation rate and C-reactive protein), and treatment outcomes were analyzed. Results: Definite microorganisms were identified in 27 patients (84.3%), with 24 (88.9%) meeting cure criteria. The cure rate was significantly higher in the detected pathogen group compared to the undetected pathogen group (88.9% vs. 80%; χ²=19.36, p<0.0001). Metagenomic next generation sequencing (mNGS) provided faster diagnosis (41.72±6.81 hours) compared to tissue culture (95.74±35.47 hours, p<0.05). The predominant causative pathogen was Mycobacterium tuberculosis, followed by Staphylococcus aureus. Significant improvements were observed in Visual Analog Scale pain scores, from a mean of 7.9 preoperatively to 1.06 at 1 year postoperatively. The Oswestry Disability Index revealed a similar trend, showing significant improvement (p<0.05). Conclusions UEDD is a viable alternative to traditional open surgery for managing IDS in high-risk patients. UEDD offers a dual therapeutic-diagnostic advantage during the initial admission phase, enabling simultaneous debridement, neurological decompression, and targeted biopsy in a single intervention. Compared with traditional tissue culture, mNGS enables rapid microbiological diagnosis and extensive pathogen coverage.

Objective To explore the value of combined ultrasound parameters, including the hepatorenal index (HRI) and renal resistance index (RRI), with cystatin C (CysC) in monitoring early acute kidney injury (AKI) after liver transplantation. Methods Perioperative data from 121 liver transplant recipients who received organs from donation after brain death were collected. The HRI and RRI of the recipients were measured on postoperative days 1-7 and at 1 month, and the CysC levels were measured on postoperative day 1. The recipients were divided into the AKI group (n=53) and the non-AKI group (n=68) based on whether AKI occurred within 7 days after operation. The data of the two groups were compared, and the ultrasound parameters before and after recovery in the AKI group were analyzed. The value of combined HRI, RRI and CysC in monitoring AKI was also analyzed. Results AKI occurred in 53 recipients, with an incidence rate of 43.8%, including 30 cases of stage 1, 18 cases of stage 2, and 5 cases of stage 3. Among them, 49 cases occurred on postoperative day 1, and 4 cases occurred on postoperative day 2. Of these, 43 cases recovered within 7 days after surgery, 8 cases recovered within 2 months after surgery, 1 case was lost to follow-up, and 1 case received renal replacement therapy. The body mass index and preoperative CysC levels were higher in the AKI group than in the non-AKI group, and the operative time was longer in the AKI group than in the non-AKI group (all P < 0.05). The HRI on postoperative day 1 was lower in the AKI group than in the non-AKI group, while the RRI and CysC levels were higher (all P < 0.05). When AKI occurred, the HRI was lower than the baseline level, and the RRI was higher than the baseline level. As AKI recovered, the HRI gradually increased, and the RRI gradually decreased. The receiver operating characteristic curve analysis showed that the sensitivity and specificity of HRI ≤ 1.12 for predicting AKI were 0.623 and 0.878, respectively, with an area under the curve (AUC) of 0.801. The sensitivity and specificity of RRI ≥ 0.65 for predicting AKI were 0.878 and 0.676, respectively, with an AUC of 0.825. The sensitivity and specificity of CysC ≥ 1.38 mg/L for predicting AKI were 0.736 and 0.882, respectively, with an AUC of 0.851 (all P<0.01). The combination of HRI and CysC (AUC=0.897, P<0.01), RRI and CysC (AUC=0.910, P<0.01), and all three parameters combined (AUC=0.934, P<0.01) were more effective than using each parameter alone. Conclusions HRI and RRI may be used to monitor the occurrence and recovery of early AKI after liver transplantation. The combination of these two parameters with CysC has a high application value in monitoring early AKI after liver transplantation.

Objective:Exploring the value of contrast enhanced ultrasound (CEUS) plus transient elastography in evaluating donor livers for C-I donors and predicting the occurrence of early allograft dysfunction (EAD).Methods:Between September 1, 2022 and August 31, 2023, the relevant clinical data were retrospectively reviewed for 75 pairs of donors and recipients. Based upon whether or not there was a postoperative onset of EAD, the recipients were assigned into two groups of EAD (16 cases) and non-EAD (59 cases) . All donors were examined by contrast-enhanced ultrasonography and FibroScan. QLAB analysis software was utilized for analyzing the results of contrast-enhanced ultrasound. Liver parenchyma at 3 cm below liver capsule was selected as a region of interest for plotting the time-intensity curve (TIC) . And the contrast-enhanced ultrasonic parameters of two groups were recorded. FibroScan transient elastography instrument was employed for quantifying liver stiffness 12 times in right lobe of donor liver and recording quantitative parameters of liver stiffness measurement (LSM) and controlled attenuation parameter (CAP) .Results:Inter-group comparison of gender, age, body mass index (BMI) and ICU length of stay showed no statistically significant differences ( P>0. 05) . However, significant differences existed in the levels of platelet [ (122. 44±85. 82) vs (197. 22± 140. 93) ×10 9/L]and cholinesterase [ (3 473. 44±1 368. 54) vs (4 252. 93±1 365. 37) U/L]within the first 24h pre-operation ( P=0. 047, P=0. 047) . Peak intensity (PKI) and area under the curve (AUC) were lower in EAD group than those in non-EAD group [ (16. 44±4. 70) dB vs 19. 85±4. 39 dB, P=0. 009; (1 366. 76±508. 10) dB·s vs (1 675. 23±498. 77) dB·s, P=0. 014]. There were statistically significant differences ( P=0. 009, P=0. 032) . Arterial-portal arrival interval (APAI) and LSM were higher in EAD group than those in non-EAD group[6. 50 (5. 00, 10.75) s vs 5. 00 (4. 00, 7. 00) s, P =0. 24; 8. 60 (6. 32, 11. 65) kPa vs 6. 10 (5. 40, 7. 90) kPa, P=0. 014]. Receiver operating characteristic (ROC) curve analysis revealed that PKI, AUC, APAI and LSM had AUC values of 0. 703, 0. 664, 0. 683 and 0. 702, respectively in predicting postoperative EAD. And combined prediction of EAD occurrence based upon these parameters had an AUC of 0. 776, a Youden index of 0. 508 with cutoff values, sensitivity and specificity of 0. 800, 0. 813 and 0. 695 respectively. Spearman' s correlation analysis revealed a negative correlation between APAI and AUC values ( r= -0. 404, P<0. 001) . Conclusions:The combination of CEUS and transient elastography can comprehensively evaluate the status of microcirculatory perfusion, fibrosis and steatosis of liver grafts from brain death donors. It offers a great predictive value for postoperative occurrence of EAD.

Medicinal plants are a valuable source of essential medicines and herbal products for healthcare and disease therapy. Compared with chemical synthesis and extraction, the biosynthesis of natural products is a very promising alternative for the successful conservation of medicinal plants, and its rapid development will greatly facilitate the conservation and sustainable utilization of medicinal plants. Here, we summarize the advances in strategies and methods concerning the biosynthesis and production of natural products of medicinal plants. The strategies and methods mainly include genetic engineering, plant cell culture engineering, metabolic engineering, and synthetic biology based on multiple "OMICS" technologies, with paradigms for the biosynthesis of terpenoids and alkaloids. We also highlight the biosynthetic approaches and discuss progress in the production of some valuable natural products, exemplifying compounds such as vindoline (alkaloid), artemisinin and paclitaxel (terpenoids), to illustrate the power of biotechnology in medicinal plants.

Objective To investigate the clinical efficacy of transurethral plasma electrotomy with rod electrode and end-to-end urethral anastomosis in the treatment of short urethral stricture.Methods 125 male patients with short urethral stricture(＜2 cm)who were admitted to our hospital from January 2014 to December 2020 were retrospectively analyzed.According to different surgical methods,they were divided into transurethral plasma resection with rod electrode(minimally invasive group)and urethral end-to-end anastomosis(open group).According to the length of urethral stricture,the minimally invasive group was divided into minimally invasive group 1(stricture length≤1 cm),minimally invasive group 2(stricture length1-2 cm),open group 1(stricture length ≤1 cm)and open group 2(stricture length 1-2 cm).The four groups were compared with each other,and the surgical success rates were compared between the four groups.Results The success rate of the minimally invasive group 1 and the open group 1 was 88.57％and 93.10％,respectively.There was no significant difference between the two groups(P＞0.05).The surgical success rate of the minimally invasive group 2 and the open group 2 was 67.86％and 90.91％,respectively.The surgical success rate of the minimally invasive group 2 was significantly lower than that of the open group 2.The difference was statistically significant(P＜0.05).The surgical success rate of minimally invasive group 1 and minimally invasive group 2 was 88.57％and 67.86％,respectively,the difference was statistically significant(P＜0.05).The success rate of operation in the open group 1 and the open group 2 was 93.10％and 90.91％,respectively,and there was no significant difference between the two groups(P＞0.05).Conclusion Transurethral plasma resection with rod electrode is preferred for urethral stricture with length≤1 cm,because the success rate of this surgery is the same as that of open surgery,and the surgical trauma is small and the recovery is fast.For 1-2 cm urethral stricture,minimally invasive surgery has less trauma and faster recovery,but open surgery has a higher success rate.The choice of surgical method needs to weigh the advantages and disadvantages and take comprehensive consideration.

Objective:To explore the genetic etiology of a child with primary hypertrophic osteoarthropathy(PHO).Methods:A child who was admitted to Zhongnan Hospital of Wuhan University on July 27, 2021 was selected as the study subject. Genomic DNA was extracted from peripheral blood samples of the child and his parents and subjected to whole exome sequencing. Suspected splicing variant was verified by Sanger sequencing of family members. In vitro function was validated through a minigene assay, whilst the suspected exonic deletion was validated by long-fragment PCR. This study was approved by the Children′s Hospital Affiliated to Zhengzhou University (Ethics No. 2023-K-011). Results:Whole exome sequencing revealed that the child has harbored compound heterozygous variants of HPGD gene, including a heterozygous deletion (Exon 3 del) derived from his father and a splicing variant (c.421+ 1G>T) derived from his mother. Long-fragment PCR verified that the child and his father had both harbored a 7 565 bp heterozygous deletion (c.218-1304_324+ 6156del), whilst the minigene assay proved that the splicing variant has resulted in skipping of exon 4. Conclusion:The heterozygous c. 218-1304_324+ 6156del deletion and the c. 421+ 1G>T splicing variant of the HPGD gene probably underlay the pathogenesis in this child. Above finding has enriched the mutational spectrum of the HPGD gene and provided a basis for genetic counseling and prenatal diagnosis for this family.

Objective:To explore the role of prognostic nutritional index (PNI) and pleural thickness in the prognostic evaluation of patients with epithelial malignant pleural mesothelioma (MPM) .Methods:In April 2022, a retrospective analysis was conducted on the data and laboratory data of 41 patients with epithelial MPM admitted to the cardiothoracic surgery department of Chuxiong Yi Autonomous Prefecture People's Hospital from January 2018 to May 2021. Univariate and multivariate analysis were used to evaluate the relationships between total survival time, clinical characteristics, PNI and pleural thickness in patients.Results:The 41 patients were mostly male (26 cases, 63.4%) , with a median age of 55 years old. The main clinical manifestations were chest pain (53.7%) , bloody pleural effusion (75.6%) , and chest pain combined with bloody pleural effusion (36.6%) . The median survival time of patients with different TNM stage, efficacy after 4 cycles of chemotherapy, PNI, maximum pleural thickness after chemotherapy (post max) , sum of post max in 3 zones after chemotherapy (post sum) were statistically different (χ 2=3.89, 14.51, 15.33, 4.33, 12.05, P<0.05) . Compared with patients with high PNI and post sum<32.26 mm, MPM patients with low PNI and post sum≥32.26 mm have higher risk of death, and the differences were statistically significant ( HR=1.52, 95% CI: 1.75-11.93, P=0.002; HR=1.70, 95% CI: 1.84-16.23, P=0.002) . Conclusion:PNI and post sum can be used to predict the prognosis of patients with epithelial MPM.