Effective annotation of in vivo drug metabolites using liquid chromatography-mass spectrometry (LC-MS) remains a formidable challenge. Herein, a metabolic reaction-based molecular networking (MRMN) strategy is introduced, which enables the "one-pot" discovery of prototype drugs and their metabolites. MRMN constructs networks by matching metabolic reactions and evaluating MS² spectral similarity, incorporating innovations and improvements in feature degradation of MS² spectra, exclusion of endogenous interference, and recognition of redundant nodes. A minimum 75% correlation between structural similarity and MS² similarity of neighboring metabolites was ensured, mitigating false negatives due to spectral feature degradation. At least 79% of nodes, 49% of edges, and 97% of subnetworks were reduced by an exclusion strategy of endogenous ions compared to the Global Natural Products Social Molecular Networking (GNPS) platform. Furthermore, an approach of redundant ions identification was refined, achieving a 10%-40% recognition rate across different samples. The effectiveness of MRMN was validated through a single compound, plant extract, and mixtures of multiple plant extracts. Notably, MRMN is freely accessible online at https://yaolab.network, broadening its applications.

Formate is an important solar fuel, with large application potential in bioconversion. Especially, the win-win collaboration is achieved when formate is applied to the cultivation of microalgae, which combines the advantages from both artificial and natural photosynthesis. However, the inhibition of formate on the photosynthetic electron transport hinders the application of formate at high concentrations. The engineering or directed evolution of the regulation pathway is a case-by-case and time-consuming strategy. Here, we developed a new strategy by introducing a Stenotrophomonas sp. strain which was isolated and identified from the long-term self-evolution process of Chlamydomonas reinhardtii for adapting to high concentrations of formate. The co-culture with the strain or the fermentation broth relieved the inhibition of formate (50 mmol/L) on C. reinhardtii and promoted the growth of the microalga. Especially, the protein content increased significantly to nearly 50% of the dried weight. In addition, the co-culture also benefited the growth of both Chlorella pyrenoidesa and Synechocystis sp. PCC 6803 exposed to formate, which indicated broader applicability of this strategy. This strategy provides the opportunity to overcome the bottleneck in the formate-mediated artificial-natural hybrid photosynthesis and to aid the development of technologies for solar energy-driven production of bulk biomass, including proteins, by carbon dioxide reduction.
Photosynthesis/physiology* ; Formates/pharmacology* ; Stenotrophomonas/growth & development* ; Microalgae/metabolism* ; Chlamydomonas reinhardtii/growth & development*

Objective To distinguish Khat from its similar plants,such as white tea and Houttuynia cordata,so as to identity Khat and its camouflage and confusing products in smuggling crime and public security anti-drug work.Methods FTIR is used to collect the infrared spectral data of plant samples,and the data are analyzed by chemometrics.The original data were preprocessed by three data processing methods,namely,SG smoothing,baseline and normalization,then the processed data were classified and reduced by PCA,finally the mathematical model was constructed by three methods:KNN,RF and SVM for classification and prediction.Results The results were expressed by confusion matrix,and the accuracy of the RF model is 96.51%.The accuracy of the SVM model is 98.84%;The accuracy of the KNN model is 93.02%.Conclusion In this experiment,FTIR was used to establish a model for rapid identification of Khat grass,the SVM model has the highest correct recognition rate and good parameters,which can be used to distinguish dry khat from its similar plants.

Purpose To detect the expression of peroxiso-mal membrane protein 4(PXMP4)in breast cancer tissues and to explore the effect of PXMP4 on the proliferation,invasion,and epithelial-mesenchymal transition(EMT)of breast cancer cells.Methods Bioinformatics and immunohistochemistry(IHC)were used to detect the expression of PXMP4 in breast cancer tissues.In breast cancer cells,Western blot was used to detect the expression of Cyclin D1,E-cadherin,vimentin and N-cadherin after knockdown and overexpression of PMXP4.The proliferation ability of breast cancer cells was analyzed by CCK-8 and plate cloning assay.Scratch healing and Transwell assay an-alyzed the migration and invasion ability of breast cancer cells.Lentivirus was used to construct a PXMP4-silenced MCF-7 cell line,and the PXMP4-silenced MCF-7 cells were injected into the subcutaneous or tail vein of mice to observe lung metastasis and the number of subcutaneous tumors.Results Bioinformat-ics and IHC showed that the expression of PXMP4 in breast cancer tissues was significantly increased(P＜0.05),and the prognosis of breast cancer patients with high expression of PXMP4 was poor(P＜0.05).The clinicopathological analysis showed that the expression of PXMP4 was correlated with tumor grade and lymph node metastasis(P＜0.05).In vitro knock-down of PMXP4 inhibited the proliferation,invasion and EMT process of breast cancer cells(P＜0.05).Conversely,overex-pression of PXMP4 promoted the proliferation,invasion and EMT process of breast cancer cells(P＜0.05).In vivo,the number of lung metastases,the size of subcutaneous tumor,and the expression of Ki67 in tumor tissue were significantly de-creased after silenced PXMP4(P＜0.05).Conclusion PXMP4 is related to tumor grading and lymph node metastasis.PXMP4 promotes proliferation,invasion and EMT process of breast cancer cells.

Objective:To evaluate the effect of pre-infusion of hypertonic saline on the postoperative cognitive function in elderly patients.Methods:This was a prospective study. Seventy-six patients of both sexes, aged≥60 yr, of American Society of Anesthesiologists Physical Status classification Ⅱ or Ⅲ, who underwent elective shoulder arthroscopic surgery under brachial plexus block combined with general anesthesia from June 2022 to January 2023 in our hospital, were selected and divided into 2 groups ( n=38 each) by the random number table method: hypertonic saline group and normal saline group. At 30 min before anesthesia induction, 3% hypertonic saline of 4 ml/kg was intravenously infused in hypertonic saline group, and normal saline 4 ml/kg was intravenously infused in normal saline group. The occurrence of intraoperative cerebral desaturation events was recorded. Venous blood samples were collected at 24 h postoperatively, and the plasma concentrations of interleukin-1beta (IL-1β), IL-6, tumor necrosis factor-alpha and S-100β were measured by enzyme-linked immunosorbent assay, and the expression of neutrophil CD11b was detected by flow cytometry. Rey auditory verbal learning test, trail making test, digit symbol substitution test, and stroop color-word test were performed at 1 day before surgery and 5 days after surgery, and the postoperative cognitive dysfunction was assessed using the Z-score method. Results:Compared with normal saline group, the concentrations of plasma IL-6, tumor necrosis factor-alpha and S-100β and expression of neutrophil CD11b were significantly decreased in hypertonic saline group, and the incidence of cognitive dysfunction and cerebral desaturation events was decreased in hypertonic saline group ( P<0.05). Conclusions:Pre-infusion of hypertonic saline can reduce inflammatory responses and improve postoperative cognitive function in elderly patients.

Objective:To analyze the effect of mucosal ligament preservation on the outcome of arthroscopic repair of meniscus injury.Methods:A total of 77 patients with knee meniscus tears caused by sports injuries who underwent arthroscopic meniscus repair or suture in Zhengzhou Orthopaedic Hospital from June 2022 to June 2023, were retrospectively analyzed. Including 45 males and 32 females, aged 38.57±13.54 years (range, 52-87 years), body mass index 24.72±4.01 kg/m 2 (range, 34.14-13.61 kg/m 2). All patients complained of knee pain, limited activity and walking weakness. The symptoms were not relieved after 1 month of conservative treatment, which seriously affected daily work and life. According to the intraoperative treatment of mucosal ligament, the patients were divided into mucosal ligament preservation group and mucosal ligament removal group. The visual analogue score (VAS), Lysholm score, and total blood loss were compared between the two groups. Results:All patients successfully completed the operation and were followed up for an average of 5.23±2.16 months (range, 3-9 months). The operation time was 47.59±16.81 min in mucosal ligament preservation group and 45.25±15.93 min in mucosal ligament removal group, and there was no significant difference between the two groups ( t=0.628, P=0.532). The total blood loss in the mucosal ligament preservation group was 246±193 ml, which was less than 343±211 ml in the mucosal ligament removal group, and the difference was statistically significant ( t=2.095, P=0.040). None of the patients received allogeneic blood transfusion. The hematocrit of the mucosal ligament preservation group was 42.48%±4.57% before operation and 39.42%±4.65% on the third day after operation, while that of the mucosal ligament removal group was 41.24%±4.16% and 38.95%±3.80%. The difference between the two groups was statistically significant ( t=0.016, P=0.004; t=0.004, P=0.016). There was no significant difference between the two groups before operation and on the third day after operation ( t=0.217, P=0.545; t=0.629, P=0.159). The preoperative VAS score of mucosal ligament preservation group was 7.25±1.10, which was higher than that of 3 months after operation (0.83±1.06), and the difference was statistically significant ( t=0.062, P<0.001). The preoperative VAS score of mucosal ligament removal group was 7.16±1.21, which was higher than that of 3 months after operation (1.05±1.13), and the difference was statistically significant ( t=0.017, P<0.001). There was no significant difference in VAS scores between the two groups before operation and at 3 months after operation ( t=0.144, P=0.740; t= 0.273, P=0.603). The preoperative Lysholm score of mucosal ligament preservation group was 31.76±7.54, which was significantly lower than that of 3 months after operation 87.30±4.12 ( t=-39.329, P<0.001); The Lysholm score of the mucosal ligament removal group was 34.13±7.32 before operation, which was lower than 89.05±4.45 at 3 months after operation, and the difference was statistically significant ( t=-40.172, P<0.001); There was no significant difference in Lysholm score between the two groups before operation and 3 months after operation ( t=1.395, P=0.167; t=1.766, P=0.081). Conclusion:The preservation of mucosal ligament in arthroscopic surgery for meniscus injury does not prolong the operation time. It can reduce the total intraoperative blood loss, and the postoperative knee function recovery is similar to that of mucosal ligament removal.

Objective:To compare the outcomes of robot-assisted laparoscopic partial nephrectomy (RAPN) and laparoscopic partial nephrectomy (LPN) in the treatment of tumors in isolated kidney, and analyze the factors influencing postoperative renal function and long-term survival in patients.Methods:A retrospective analysis was conducted on clinical data of 67 patients with tumors in isolated kidney who underwent surgery at the Chinese PLA General Hospital from November 2010 to January 2022. There were 48 males and 19 females, with an average age of (58.6±10.1) years old. The patients were divided into RAPN group (43 cases) and LPN group (24 cases) based on the surgical approach. The RAPN group had a higher R.E.N.A.L. score than the LPN group [(8.7±1.5) vs. (7.9±1.7), P=0.042]. There were no statistically significant differences between the two groups in terms of age [(57.4±10.2) years old vs. (60.9±9.8) years old, P=0.185], body mass index (BMI) [(25.7±3.5) kg/m 2 vs. (25.1±3.6) kg/m 2, P=0.518], and preoperative serum creatinine [(102.9±31.6) μmol/L vs. (102.3±22.4) μmol/L, P=0.930]. Twelve cases underwent hypothermic treatment during surgery, with 9 cases(20.9%) in the RAPN group and 3 cases(12.5%) in the LPN group( P=0.596). Surgical time, intraoperative warm ischemia time, intraoperative blood loss, postoperative fasting time, perioperative complication rate, postoperative serum creatinine, and other indicators were compared between the two groups. Multiple linear regression analysis was used to identify factors affecting postoperative serum creatinine. Kaplan-Meier curves were employed to analyze patient prognosis, and log-rank tests were performed to compare the differences between the two groups. Multiple Cox regression analysis was used to identify factors influencing patient prognosis. Results:All surgeries were completed successfully with negative pathological margins. There were no statistically significant differences between the RAPN and LPN groups in terms of surgical time [(136.6±47.6) min vs. (125.3±34.4) min, P=0.311], intraoperative ischemia time [23.0 (16.0, 30.0) min vs. 19.0 (13.5, 27.5) min, P =0.260], intraoperative blood loss [50.0 (50.0, 100.0) ml vs. 50.0 (22.5, 100.0) ml, P=0.247], postoperative hospital stay [(6.6±3.5) days vs. (7.7±4.2) days, P=0.244], time to drain removal [4(3, 5) days vs. 5(3, 6) days, P =0.175], postoperative fasting time [(2.1±0.7) days vs. (2.2±1.0) days, P=0.729], perioperative complication rate [18.6% (8/43) vs. 16.7% (4/24), P=1.000], postoperative serum creatinine [145.2 (128.3, 191.3) μmol/L vs. 157.8 (136.2, 196.3) μmol/L, P =0.229], and pathological staging [T 1a/T 1b/T 2a/T 3a/T 4 stage: 32/7/1/3/0 case vs. 17/5/0/1/1 case, P=0.804]. Kaplan-Meier survival curves showed that the total survival rates at 1, 3, and 5 years after surgery were 94.7%, 84.9%, and 84.9% for the RAPN group, and 100.0%, 95.5%, and 95.5% for the LPN group, with no statistically significant difference in the log-rank test ( P=0.116). Excluding 10 patients with preoperative tumor metastasis (7 in the RAPN group and 3 in the LPN group), the progression-free survival rates at 1, 3, and 5 years after surgery were 84.8%, 81.1%, and 81.1% for the RAPN group, and 100.0%, 95.0%, and 90.0% for the LPN group, with no statistically significant difference in the log-rank test ( P =0.142). Multiple linear regression analysis showed that the use of hypothermic treatment during surgery significantly reduced postoperative serum creatinine ( B=-72.191, P=0.048). Multiple Cox regression analysis revealed that BMI ( HR=0.743, P=0.044), pathological T stage ( HR=4.235, P=0.018), and preoperative metastasis ( HR=18.829, P=0.035) were independent factors affecting patient overall survival time. A smaller BMI, higher pathological stage, and preoperative metastasis were associated with poorer prognosis. Conclusions:Despite the higher R. E.N.A.L. score and greater surgical difficulty in the RAPN group, RAPN achieved similar perioperative and prognostic results as the LPN, indicating RAPN advantages in treating tumors in isolated kidney. Appropriate intraoperative hypothermic treatment can better protect postoperative renal function. BMI, pathological T stage, and preoperative metastasis are independent factors affecting overall survival time.

Objective To identify the retention time of semen(stains)inside and outside the cadaver using Fourier Transform Infrared(FTIR)spectroscopy and chemometrics.Methods A simulated environment inside the porcine vagina was used to retain fresh semen for 0～168 hours inside and outside the body.Infrared spectroscopy data were collected,and chemometric methods including Principal Components Analysis(PCA)and Partial Least Squares(PLS)regression models were established.Results The infrared spectrum peak positions of semen(stains)inside and outside the body did not show significant changes over time.The PCA results showed good clustering relationships,and the predicted deviation value in the PLS regression model was 15.786 3 hours,with an R2 value of 0.922 4.The characteristic wavenumbers selected by the CARS algorithm were in the wavelength ranges of 1 000～1 180 cm-1,1 430～1 480 cm-1,and 1 700～1 760 cm-1.Conclusion Fourier Transform Infrared spectroscopy has the potential to identify the retention time of semen(stains)inside and outside the body,and there are differences in the degradation trends of semen(stains)inside and outside the body.

Objective:To investigate the clinical effect of arthroscopic release through the peritrochanteric compartment from inside-out in the treatment of lateral snapping hip.Methods:A total of 38 patients (70 hips) with lateral snapping hip who underwent arthroscopic surgery in Department of Sports Medicine, Zhengzhou Orthopaedics Hospital Affiliated to Henan University from December 2016 to December 2022 were retrospectively analyzed. There were 13 males and 25 females, aged 27.8±7.6 years (range, 15-45 years). Among them 6 cases of unilateral and bilateral 32 cases. According to the different release approaches, they were divided into outside-in group (37 hips) and inside-out group (33 hips). Visual analogue scale (VAS) was used to evaluate postoperative hip pain, and gluteal muscle contracture disability scale (GDS) was used to evaluate postoperative effectiveness. Postoperative hip function was evaluated by modified Harris hip score (mHHS).Results:All patients in the two groups successfully completed the operation and were followed up for 8.23±1.25 months (range, 6-12.8 months). The hip VAS scores in the inside-out group were 4.0 (3.5, 6.0), 5.0 (4.0, 6.0), and 1.0 (0.5, 1.0) before surgery, 1 week after surgery, and 3 months after surgery, respectively, and those in the outside-in group were 4.0 (3.5, 6.0), 5.0 (4.0, 6.0), and 1.0 (0.5. 1.0), and the differences between the two groups of patients in the within group comparison of hip VAS before and after surgery were statistically significant ( P<0.05). The hip VAS at 3 months postoperatively was smaller than that before surgery and at 1 week postoperatively, and the differences were statistically significant ( P<0.05). There were no significant differences between groups at different time points ( P>0.05). The GDS scores in the inside-out group were 52.30±8.19, 75.03±6.41, and 92.30±4.61 preoperatively, 1 week postoperatively, and 3 months postoperatively, respectively, and in the outside-in group were 51.97±6.64, 75.41±7.10, and 91.32±4.18, respectively, and the differences in the quantitative GDS function scores of the hips in the pre- and postoperative periods of the patients in both groups were statistically significant ( P<0.05). The differences in the quantitative GDS function scores within the groups were statistically significant ( P<0.05), and the quantitative GMC function scores of the hip joints at 3 months after surgery were greater than those of the preoperative period and 1 week after surgery, and the differences were statistically significant ( P<0.05). There were no significant differences between groups at different time points ( P>0.05). Differences in mHHS within group comparisons before and after surgery were statistically significant in both groups ( P<0.05), and mHHS at 3 months postoperatively was greater than preoperatively and 1 week postoperatively, with statistically significant differences ( P<0.05). The mHHS at 1 week postoperatively in the inside-out group was 79.03±7.37, which was greater than that in the outside-in group 71.32±10.02, and the difference was statistically significant ( t=3.63, P<0.001). Conclusion:Arthroscopic release through the peritrochanteric compartment from inside-out in the treatment of lateral snapping hip can effectively reduce hip pain and improve hip function, with satisfactory clinical results, and can be used as an alternative treatment to inward-to-outward release.

Objective:To evaluate the effect of esketamine on hippocampal neuronal necroptosis in aged rats with postoperative cognitive dysfunction.Methods:One hundred and twenty SPF-grade healthy male Sprague-Dawley rats, aged 22 months, weighing 550-600 g, were divided into 4 groups ( n=30 each) using a random number table method: control group (group C), postoperative cognitive dysfunction group (group P), postoperative cognitive dysfunction+ esketamine group (group PE), and esketamine group (group CE). Rats received exploratory laparotomy under sevoflurane anesthesia, and esketamine 10 mg/kg and the equal volume of 0.9% sodium chloride were intraperitoneally injected at the end of surgery once a day for 6 consecutive days in group P and group PE, respectively. Rats received no anesthesia and surgery, and esketamine 10 mg/kg and the equal volume of 0.9% sodium chloride were intraperitoneally injected at the end of surgery once a day for 6 consecutive days in group CE and group C, respectively. Morris water maze test was performed at 7th day after surgery. The escape latency, times of crossing the original platform and time spent in the original platform quadrant were recorded. The rats were sacrificed at the end of Morris water maze test, and the hippocampal tissues were collected for determination of the rate of necroptosis and cytosolic Ca 2+ concentrations (by flow cytometry) and expression of mixed lineage kinase domain-like protein (MLKL), phosphorylated MLKL (p-MLKL), receptor-interacting protein kinase-3 (RIPK3), phosphorylated RIPK3 (p-RIPK3), receptor-interacting protein kinase-1 (RIPK1) and phosphorylated RIPK1 (p-RIPK1) (by Western blot). Results:Compared with group C, the escape latency was significantly prolonged, the times of crossing the original platform were decreased, the time spent in the original platform quadrant was shortened, the necroptosis rate of hippocampal neurons and cytosolic Ca 2+ concentrations were increased, and the expression of MLKL, p-MLKL, RIPK3, p-RIPK3, RIPK1 and p-RIPK1 was up-regulated in group P and group PE ( P<0.05). Compared with group P, the escape latency was significantly shortened, the times of crossing the original platform were increased, the time spent in the original platform quadrant was prolonged, the necroptosis rate of hippocampal neurons and cytosolic Ca 2+ concentrations were decreased, and the expression of MLKL, p-MLKL, RIPK3, p-RIPK3, RIPK1 and p-RIPK1 was down-regulated in group PE ( P<0.05). Conclusions:The mechanism by which esketamine attenuates postoperative cognitive dysfunction may be related to inhibition of necroptosis in hippocampal neurons of aged rats.