In protein engineering, while computational models are increasingly used to predict mutation effects, their evaluations primarily rely on high-throughput deep mutational scanning (DMS) experiments that use surrogate readouts, which may not adequately capture the complex biochemical properties of interest. Many proteins and their functions cannot be assessed through high-throughput methods due to technical limitations or the nature of the desired properties, and this is particularly true for the real industrial application scenario. Therefore, the desired testing datasets, will be small-size (∼10-100) experimental data for each protein, and involve as many proteins as possible and as many properties as possible, which is, however, lacking. Here, we present VenusMutHub, a comprehensive benchmark study using 905 small-scale experimental datasets curated from published literature and public databases, spanning 527 proteins across diverse functional properties including stability, activity, binding affinity, and selectivity. These datasets feature direct biochemical measurements rather than surrogate readouts, providing a more rigorous assessment of model performance in predicting mutations that affect specific molecular functions. We evaluate 23 computational models across various methodological paradigms, such as sequence-based, structure-informed and evolutionary approaches. This benchmark provides practical guidance for selecting appropriate prediction methods in protein engineering applications where accurate prediction of specific functional properties is crucial.

Osteoporosis is a prevalent skeletal condition characterized by reduced bone mass and strength, leading to increased fragility. Buqi-Tongluo (BQTL) decoction, a traditional Chinese medicine (TCM) prescription, has yet to be fully evaluated for its potential in treating bone diseases such as osteoporosis. To investigate the mechanism by which BQTL decoction inhibits osteoclast differentiation in vitro and validate these findings through in vivo experiments. We employed MTS assays to assess the potential proliferative or toxic effects of BQTL on bone marrow macrophages (BMMs) at various concentrations. TRAcP experiments were conducted to examine BQTL's impact on osteoclast differentiation. RT-PCR and Western blot analyses were utilized to evaluate the relative expression levels of osteoclast-specific genes and proteins under BQTL stimulation. Finally, in vivo experiments were performed using an osteoporosis model to further validate the in vitro findings. This study revealed that BQTL suppressed receptor activator of NF-κB ligand (RANKL)-induced osteoclastogenesis and osteoclast resorption activity in vitro in a dose-dependent manner without observable cytotoxicity. The inhibitory effects of BQTL on osteoclast formation and function were attributed to the downregulation of NFATc1 and c-fos activity, primarily through attenuation of the MAPK, NF-κB, and Calcineurin signaling pathways. BQTL's inhibitory capacity was further examined in vivo using an ovariectomized (OVX) rat model, demonstrating a strong protective effect against bone loss. BQTL may serve as an effective therapeutic TCM for the treatment of postmenopausal osteoporosis and the alleviation of bone loss induced by estrogen deficiency and related conditions.
Animals ; NFATC Transcription Factors/genetics* ; Drugs, Chinese Herbal/pharmacology* ; Ovariectomy ; Osteoclasts/metabolism* ; Female ; Osteogenesis/drug effects* ; Rats, Sprague-Dawley ; Rats ; NF-kappa B/genetics* ; Osteoporosis/genetics* ; Signal Transduction/drug effects* ; Bone Resorption/genetics* ; Cell Differentiation/drug effects* ; Humans ; RANK Ligand/metabolism* ; Mitogen-Activated Protein Kinases/genetics* ; Transcription Factors

Objective:To discuss the effect of azathioprine(AZA)on ferroptosis in spermatocytes of the mice induced by reduced glutathione(GSH)peroxidase 4 inhibitor RSL3,and to clarify the potential mechanism.Methods:The spermatogonia GC-2 cells of the mice were randomly divided into control group(no treatment),RSL3 group(treated with 10 nmol·L-1 RSL3 for 24 h),RSL3+ferroptosis inhibitor(Ferrostatin-1,Fer-1)group(treated with 10 nmol·L-1 RSL3 for 24 h+2 μmol·L-1 Fer-1 for 12 h),RSL3+low dose of AZA group(treated with 10 nmol·L-1 RSL3 for 24 h+5 μmol·L-1 AZA for 12 h),RSL3+medium dose of AZA group(treated with 10 nmol·L-1 RSL3 for 24 h+10 μmol·L-1 AZA for 12 h),and RSL3+high dose of AZA group(treated with 10 nmol L-1 RSL3 for 24 h+20 μmol·L-1 AZA for 12 h).The MTT method was used to detect the activities of the GC-2 cells in various groups after treated with different concentrations of AZA and RSL3;the GSH and GSSG levels in the GC-2 cells were detected by GSH and oxidized glutathione(GSSG)detection kits;the malondialdehyde(MDA)levels in the GC-2 cells in various groups were detected by MDA detection kit;Western blotting method was used to detect the expression levels of long-chain acyl-CoA synthetase 4(ACSL4),heme oxygenase-1(HO-1),and glutathione peroxidase 4(GPX4)proteins in the cells in various groups;immunofluorescence staining was used to detect the expressions of ACSL4 protein in the cells in various groups.Results:Compared with control group,the differences in activities of the GC-2 cells in 5,10,and 20 μmol·L-1 AZA groups had no significant differences(P＞0.05),while the activities of the GC-2 cells in 30 and 40 μmol·L-1 AZA groups were significantly decreased(P＜0.01);therefore,the AZA concentration was selected to be within 20 μmol·L-1.Compared with control group,the differences of the activities of the GC-2 cells in 1,5,and 10 nmol·L-1 RSL3 groups had no significant differences(P＞0.05),while the activities of the GC-2 cells in 50,100,500,and 1 000 nmol·L-1 RSL3 groups were significantly decreased(P＜0.05 or P＜0.01);therefore,the RSL3 concentration was set to be within 10 nmol·L-1.The GSH and MDA detection kits results showed that compared with control group,the levels of GSSG and MDA in the GC-2 cells in RSL3 group were significantly increased(P＜0.05),while the GSH levels were significantly decreased(P＜0.05);compared with RSL3 group,the levels of GSSG and MDA in the GC-2 cells in RSL3+Fer-1 group and RSL3+AZA group were significantly decreased(P＜0.01),while the GSH levels were significantly increased(P＜0.01).The Western blotting results showed that compared with control group,the expression levels of ACSL4 and HO-1 proteins in the GC-2 cells in RSL3 group were significantly increased(P＜0.05),while the expression level of GPX4 protein was significantly decreased(P＜0.01);compared with RSL3 group,the expression levels of GPX4 protein in the GC-2 cells in RSL3+Fer-1 group and RSL3+AZA group were significantly increased(P＜0.05),while the expression levels of ACSL4 and HO-1 proteins were significantly decreased(P＜0.01).The immunofluorescence staining results showed that compared with control group,the expression amount of ACSL4 protein in the GC-2 cells in RSL3 group was significantly increased,and compared with RSL3 group,the expression amounts of ACSL4 protein in the cells in RSL3+Fer-1 group and RSL3+AZA group were significantly decreased.Conclusion:AZA can alleviate the ferroptosis-induced by RSL3 in spermatocytes of the mice.

Objective:To discuss the protective effect of sodium butyrate(NaB)on acute liver injury in the mice induced by lipopolysaccharide(LPS)combined with D-galactosamine(D-Gal),and to clarify its mechanism.Methods:Thirty male Kunming mice were randomly divided into control group,model group,and NaB group,and there were 10 mice in each group.The mice in NaB group were given 200 mg·kg-1·d-1 NaB,while the mice in control group and model group were given an equal volume of sterile water.The mice in model group and NaB group were intraperitoneally injected with 20 μg·kg-1 LPS and 600 mg·kg-1 D-Gal to induce the acute liver injury models.The body weights and liver weights of the mice in various groups were detcted,and the liver index was calculated.HE staining was used to observe the pathomorphology of liver tissue of the mice in various groups;kits were used to detect the activities of alanine aminotransferase(ALT)and aspartate aminotransferase(AST)in serum,and the activities of total superoxide dismutase(T-SOD)and catalase(CAT),and the levels of malondialdehyde(MDA)in liver tissue of the mice in various groups;Western blotting method was used to detect the expression levels of nuclear factor E2-related factor 2(Nrf2)and heme oxygenase-1(HO-1)proteins in liver tissue of the mice in various groups.Results:There were no significant differences in body weights of the mice among various groups(P>0.05).Compared with control group,the liver index of the mice in model group was significantly increased(P<0.01).Compared with model group,the liver index of the mice in NaB group was significantly decreased(P<0.01).The HE staining results showed that the liver tissue of the mice in control group exhibited normal structure,with clear boundaries of hepatocytes,consistent size,radially arranged around the central vein,and the nucleus located in the center of the cells;in model group,the arrangement of hepatocytes was disordered,the cells were swollen,there were multiple foci of hepatocellular necrosis,inflammatory cell infiltration,and hemorrhage;compared with model group,the cells in NaB group showed improved hepatocellular structure and reduced inflammatory infiltration.Compared with control group,the activities of ALT and AST in serum of the mice in model group were significantly increased(P<0.01);compared with model group,the activities of ALT and AST in serum of the mice in NaB group were significantly decreased(P<0.05 or P<0.01).Compared with control group,the activities of T-SOD and CAT in liver tissue of the mice in model group were significantly decreased(P<0.01),and the level of MDA was significantly increased(P<0.01);compared with model group,the activities of T-SOD and CAT in liver tissue of the mice in NaB group were significantly increased(P<0.05 or P<0.01),and the level of MDA was significantly decreased(P<0.01).The Western blotting results showed that compared with control group,the expression levels of Nrf2 and HO-1 proteins in liver tissue of the mice in model group were significantly decreased(P<0.05);compared with model group,the expression levels of Nrf2 and HO-1 proteins in liver tissue of the mice in NaB group were significantly increased(P<0.01).Conclusion:NaB has a protective effect on LPS/D-Gal induced acute liver injury in the mice,and its mechanism may be related to the upregulation of the expressions of Nrf2 and HO-1 proteins and the increas of the activity of oxidant enzyme in liver tissue by NaB,thereby reduces the liver oxidative stress level of liver.

Objective To compare the effectiveness of different β-blockers for preventing postoperative atrial fibrillation (POAF) after cardiac surgery. Methods Databases of PubMed, Science Direct, Web of Science, The Cochrane Library, SinoMed, CKNI, VIP, WanFang were searched by the computer from inception to April 31, 2022 to collect randomized controlled studies on the comparison of the effectiveness of different β-blockers for preventing POAF. Two investigators independently screened the literature and extracted information. The quality of the literature was evaluated using Cochrane bias risk tool, and RevMan 5.3 and STATA 17.0 were used for meta-analysis. Results A total of 17 randomized controlled studies with 3 290 patients were included. Direct meta-analysis showed that landiolol and metoprolol were more effective than placebo (P≤0.001), and carvedilol was more effective than metoprolol in preventing the development of POAF (P<0.001). Network meta-analysis showed that landiolol, carvedilol and metoprolol were more effective than placebo in preventing the incidence of POAF (P<0.05). Landiolol and carvedilol were more effective than metoprolol, and carvedilol were more effective than nebivolol (P<0.05). The surface under the cumulative ranking curve from high to low were carvedilol, landilol, propranolol, atenolol, metoprolol, and nebivolol. Conclusion Carvedilol and landilol have different degrees of improvement in the occurrence of POAF, and carvedilol has the best preventive effect. More studies are required to verify the strength of evidence due to the limited sample size.

Objective To observe the value of FOLFOX-hepatic arterial infusion chemotherapy(HAIC)combined with programmed cell death receptor-1(PD-1)inhibitor+targeted drug for treating China liver cancer staging(CNLC)stageⅢa hepatocellular carcinoma(HCC).Methods Sixty-one patients with CNLC stage Ⅲa HCC who underwent PD-1 inhibitor+targeted drug treatment were retrospectively enrolled and divided into observation group(n=30)and control group(n=31)based on whether received FOLFOX-HAIC treatment or not.The general information,treatment strategy,adverse reactions and therapeutic effects were compared between groups,and the value of treatment strategy in observation group was analyzed.Results No significant difference of general information nor PD-1 inhibitor+targeted drug strategy was found between groups(both P＞0.05).Among 1-2 grade adverse reactions,the incidence of nausea,vomiting and abdominal pain in observation group were higher than those in control group(both P＜0.05),while no significant difference of the other 1-2 grade nor 3 grade adverse reactions was observed(all P＞0.05).The objective response rate(ORR),progression free survival(PFS)and overall survival(OS)of observation group were all higher than those of control group(all P＜0.05).Conclusion FOLFOX-HAIC combined with PD-1 inhibitor+targeted drug was more effective for treating CNLC stage Ⅲa HCC with acceptable safety.

Objective To observe the drainage pathways of interstitial fluid(ISF)and substance clearance pattern in rat brain with fluorescence tracing imaging and treacer-based MRI.Methods Thirty-three male SD rats were randomly divided into fluorescence tracing group(F group,n=18)and treacer-based MRI group(MRI group,n=15),then further divided into thalamic,hippocampal and caudate nucleus subgroups,respectively.Evans blue was injected to rats in F group,and cardiac perfusion was performed after injection,then brain tissue was harvested,and frozen sections were made to observe the drainage pathways of IFS in different subgroups.MRI was performed on rats in MRI group before and after injection of gadolinium-diethylenetriamine pentaacetic acid(Gd-DTPA)to observe signal intensity in ROI of brain regions in different subgroups,the signal unit ratio was calculated,and the changing trend was explored.Results ISF in thalamus,hippocampus and caudate nucleus had different dominant drainage pathways,and the time of tracer reached to adjacent brain regions and whole brain in F group were different.In MRI group,within 4 h after injection of Gd-DTPA,there were differences in direction and clearance rate among tracer in thalamus,hippocampus and caudate nucleus,mainly manifesting as the tracer in thalamus and hippocampus drained to the ipsilateral cortex and lateral ventricle,while the tracer in the caudate nucleus diffused to the cortex and midbrain,and there were differences of the peak time of tracer signal among adjacent drainage brain regions.Conclusion Fluorescence and MR dual-mode imaging showed that there were differences in the dominant drainage pathways of IFS and clearance rates of small molecule substances among hypothalamus,hippocampus and caudate nucleus of rats.

Through the expert questionnaire (Delphi Indagate) to form Traditional Chinese Medicine (TCM) Healthy Lifestyle Guiding Principles.We conducted two rounds of surveys of TCM experts in the field of preventive health care on the basis of previous literature research.This study was aimed to form the framework in order to entry to the TCM Healthy Life-style Guiding Principles.The results showed that the positive coefficient of experts in two rounds of investigation were 84.31％ and 100％,respectively.The authoritative decree were both more than 0.7.The expert opinion score mean median was 9.21 and 9.085.Median mean values were 7.965 and 7.925.The median scores of full marks were higher than 65.12％ and 66.67％.The minimum coefficients of variation were 0.11 and 0.10.It was concluded that in the investigation and study,the enthusiasm of the experts,professional standards and credibility were high.Expert opinion is more concentrated.Expert opinion is well coordinated and gradually converges through two rounds of surveys.Through the Delphi indagate,the basic framework and items of TCM guiding principles of healthy lifestyle were established,which laid the foundation for further standardizing the guiding principles of TCM in healthy lifestyle.

Objective To explore the value of three-dimensional DSA (3D-DSA) in displaying the location of the origin of uterine artery.Methods A total of 90 female patients underwent uterine artery (UA) embolization were enrolled.The bilateral internal iliac artery catheterization were performed by 3D-DSA,then the images were reconstructed in every 5 degree interval to choose the optimum range of viewing angle.The origination and the degree of the origin artery and UA were calculated.The distance between the origin of UA and superior glutea artery which was identified as the locating point was measured.Results Bilateral and contralateral oblique position of ＞30°-60°were the optimal projection positions of UA.Totally 64.44％ (116/180) of UA originated from the anterior trunk of internal iliac artery,18.33％ (33/180) originated from the inferior gluteal trunk,9.44 ％ (17 / 180) originated from the internal pundenal artery,5.56 ％ (10 / 180) originated from internal iliac artery,and 2.22％ (4/180) originated from the superior gluteal artery;10.56％ (19/180) of the angle of the origin artery and UA were 0-30°,38.89％ (70/180) were ＞30°-60°,41.11％ (74/180) were ＞60°-90°,4.44％ (8/180) were＞90°-120°,2.78％ (5/180) were＞120°-150°,2.22％ (4/180) were＞150°-180°.Distance between the origin of UA and superior gluteal artery was 3.04-18.31 mm,average was (11.71±4.28)mm.Conclusion 3D-DSA can clearly display the origination,viewing angle and the distance away from superior gluteal artery.

Objective To explore the impact of sulfentanyl on sufentanil epidural block during abdominal panhysterectomy. Methods 90 patients scheduled for panhysterectomy were randomly divided into three groups. Tthe control group received epidural administration of 1% ropivacaine of 0.2 mL/kg after 2% idocaine of 3 mL , while the study group 1 received 10μg sufentanil and the study group 2 received 20μg sufentanil in addition to the medications used in the control group. The anesthetic effect, changes in vital signs, and incidence of adverse reactions were compared among the three groups. Results In group S1 and group S2, the onset of epidural anesthesia was faster , time to the highest plane of sensory blockade and time to degree 3 in the Bromag scores were faster , duration of sensory blockade was longer , and OAA/S score was better , as compared with group D , with significant statistical significances (P<0.01);and the effect was better in group S2 than in group S1. There was no difference among the three groups in adverse reactions. MAP , HR and SPO2 were lower in groups S1 and S2 than in group D during the procedure, with a statistical difference (P<0.05). Conclusions Proper dose of sufentanil plays a positive role in ropivacaine epidural block during panhysterectomy , not only increases the onset of anesthesia, but also makes the anesthestic effect better, and has higher safety It is worth popularizing clinically.