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:The variability of the apnea-hypopnea index（AHI） measured in the first and second halves of the night is significant in patients with obstructive sleep apnea hypopnea syndrome（OSAHS）. This variation may be related to fluid redistribution caused by the supine position during sleep. Methods:Eighty-nine adult subjects were enrolled. Circumferences（neck, chest, waist, and calf） were measured before sleep onset and upon awakening. Polysomnography（PSG） was performed, and the night was divided into two halves based on the midpoint of total sleep time to calculate AHI for each half. The correlation between changes in AHI and changes in circumferences was analyzed. Results:Twenty simple snorers and sixty-nine OSAHS patients were included, with a median AHI of 22.6（11.8, 47.3） events/hour. Compared to pre-sleep measurements, there was no significant change in neck circumference upon awakening in the control group（P=0.073）, while reductions were observed in the other three measurements（P=0.006, P=0.038, P<0.001）. In the OSAHS group, neck circumference increased（P<0.001）, and reductions were noted in the other three measurements（P<0.001 for all）, with the most significant change observed in calf circumference 40.0（37.1, 42.0） cm to 38.0（35.8, 40.5） cm. Compared to the first half of the night, total AHI, supine AHI, and NREM AHI significantly decreased in the second half（P=0.010, P=0.031, P=0.001）, while no significant changes were observed in lateral AHI and REM AHI（P=0.988, P=0.530）. Further analysis revealed a significant relationship between increased chest circumference and decreases in NREM AHI, supine AHI, and supine NREM AHI（P=0.036, P=0.072, P=0.034）, as well as between decreased lateral position AHI and increased waist circumference（P=0.048）. Additionally, this study found a negative correlation between changes in calf circumference and changes in AHI（R=-0.24, P=0.048）, while neck circumference changes positively correlated with changes in AHI（R=0.26, P=0.03）. Conclusion:In OSAHS patients during the second half of sleep compared to before sleeping, chest circumference, waist circumference, and calf circumference decrease while neck circumference increases; total AHI, supine position AHI, and NREM period AHI decrease; increases in chest circumference are associated with decreases in NREM period AHI, supine position AHI, supine position NREM period AHI. There is nocturnal variability in AHI among OSAHS patients that may be associated with fluid shifts during sleep.
Humans ; Sleep Apnea, Obstructive/physiopathology* ; Male ; Female ; Polysomnography ; Fluid Shifts/physiology* ; Adult ; Middle Aged ; Neck ; Severity of Illness Index ; Sleep/physiology* ; Snoring/physiopathology*

Anxiety disorder is a major symptom of autism spectrum disorder (ASD) with a comorbidity rate of ~40%. However, the neural mechanisms of the emergence of anxiety in ASD remain unclear. In our study, we found that hyperactivity of basolateral amygdala (BLA) pyramidal neurons (PNs) in Shank3 InsG3680 knock-in (InsG3680^+/+) mice is involved in the development of anxiety. Electrophysiological results also showed increased excitatory input and decreased inhibitory input in BLA PNs. Chemogenetic inhibition of the excitability of PNs in the BLA rescued the anxiety phenotype of InsG3680^+/+ mice. Further study found that the diminished control of the BLA by medial prefrontal cortex (mPFC) and optogenetic activation of the mPFC-BLA pathway also had a rescue effect, which increased the feedforward inhibition of the BLA. Taken together, our results suggest that hyperactivity of the BLA and alteration of the mPFC-BLA circuitry are involved in anxiety in InsG3680^+/+ mice.
Animals ; Prefrontal Cortex/metabolism* ; Basolateral Nuclear Complex/metabolism* ; Mice ; Anxiety/metabolism* ; Nerve Tissue Proteins/genetics* ; Male ; Gene Knock-In Techniques ; Pyramidal Cells/physiology* ; Mice, Transgenic ; Neural Pathways/physiopathology* ; Mice, Inbred C57BL ; Microfilament Proteins

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.