OBJECTIVE To evaluate the clinical value of anlotinib in third-line treatment for patients with advanced non-small cell lung cancer （NSCLC） through real-world data. METHODS Clinical data of patients with advanced NSCLC who received treatment at the First Affiliated Hospital of Anhui University of Chinese Medicine from February 2021 to December 2024 were retrospectively collected. They were divided into anlotinib group （27 cases， receiving anlotinib therapy） and immunotherapy group （22 cases， receiving immunotherapy agents alone or in combination with chemotherapy drugs） according to treatment regimens. The progression-free survival （PFS） and overall survival （OS） of patients were compared between the two groups， and the occurrence of adverse drug reactions during the treatment period was recorded. Using a partitioned survival model， an economic evaluation of the two treatment regimens was conducted with a cost-utility analysis approach from the perspective of the healthcare system. RESULTS The median PFS and OS of patients in the anlotinib group were 5.93 months and 11.27 months， respectively； the median PFS and OS of patients in the immunotherapy group were 5.33 months and 9.77 months， respectively； the difference was not statistically significant （P＞0.05）. There was no statistical difference in the total incidence of adverse drug reactions and grade 3-4 serious adverse drug reactions between the two groups （P＞0.05）. Compared with the immunotherapy group， the incremental cost-effectiveness ratio of the anlotinib group was 1 806 724.60 yuan/quality-adjusted life year （QALY）， which was significantly higher than three times China’s per capita gross domestic product in 2024 （287 247 yuan/QALY）. CONCLUSIONS For third-line treatment of advanced NSCLC patients， the efficacy of anlotinib is no worse than that of immunotherapy alone or in combination with chemotherapy drugs， and the safety of the two groups is comparable. However， anlotinib is not cost-effective.

Objective To explore the effects of two methods of video laryngoscope insertion on the success rate of tracheal intubation and postoperative pharyngeal complications.Methods We selected 80 elective surgerypatients with American Society of Anesthesiologists(ASA)classification Ⅰ to Ⅱ who had no difficult airway in preoperative assessment under general anesthesia with orotracheal intubation.They were aged 18 to 65 years old and there were no restrictions on height and weight.The patients were randomly divided into a group with laryngoscope inserted through the middle of the mouth(group M)and a group with laryngoscope inserted through the right corner of the mouth(group C),with 40 cases in each group.Observe the laryngoscope exposure grading,oropharyngeal mucosal bleeding,first-time intubation success rate,and occurrence of pharyngeal pain and hoarseness within 48 hours after surgery in both groups;record the patients'systolic blood pressure(SBP),diastolic blood pressure(DBP),and heart rate(HR)among admission to the room(T0),after giving anesthesia drugs(T1),immediately after intubation(T2),1 min after intubation(T3),and 3 min after intubation(T4).Results The SBP of group M was lower than that of group C at T3(P＜0.05);There was no statistically significant difference in laryngoscope exposure grading between the two groups(P＞0.05).The intubation time,oropharynx mucosal bleeding and the cases of sore throat within 48 hours of group M were less than those in group C(P＜0.05).The success rate of primary intubation in group M was higher than that in group C(P＜0.05).Conclusion Inserting a video laryngoscope through the middle of the mouth can reduce oropharyngeal mucosal bleeding,improve the success rate of intubation,and reduce the incidence of postoperative pharyngeal complications.

Background and purpose:Colorectal cancer(CRC)is one of the major malignant tumors threatening human health worldwide,with long-term high incidence and mortality rate.Potassium channel modulatory factor 1(KCMF1)is a member of the E3 ubiquitin ligase family.It binds to target proteins through the RING domain and participates in the regulation of a variety of biological processes in vivo.However,the function of KCMF1 in CRC remains unclear.This study aimed to investigate the expression level of E3 ubiquitin ligase KCMF1 in colorectal tumor,and to explore the effects of KCMF1 on the proliferation of CRC cells and its underlying molecular mechanism.Methods:The The Cancer Genome Atlas(TCGA)and Genotype-Tissue Expression(GTEx)databases were used to analyze the expression level of KCMF1 in CRC tissues and adjacent tissues and the association between the KCMF1 expression and the prognosis of CRC patients.Furthermore,immunohistochemical staining was performed to detect the protein level of KCMF1 in 90 paired human CRC tissues and adjacent non-tumor tissues.Lentiviral shRNA delivery system was employed to specifically target the KCMF1 gene(shKCMF1)in HCT116 and HCT15 CRC cell lines.The effects of KCMF1 knockdown on cell proliferation,apoptosis and cell cycle distribution were assessed by methyl thiazoyl terazolium(MTT)assay,colony formation assay,Western blot and flow cytometry.Changes in the transcriptional profile in HCT116 cells upon KCMF1 knockdown were identified by RNA sequencing(RNA-Seq),and the affected signaling pathways were evaluated by bioinformatics analysis.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR),Western blot,luciferase reporter assay and cell immunofluorescence assay were utilized to validate the alteration of the affected signaling pathway.Results:The TCGA and GTEx databases and IHC results showed that the mRNA and protein expression levels of KCMF1 in CRC tissues were significantly upregulated compared with adjacent tissues(P＜0.01).KCMF1 expression level was negatively correlated with the survival time of patients with CRC(P＜0.01),and was positively associated with CRC clinical stage(P＜0.05).Compared with control cells,KCMF1 knockdown significantly inhibited the proliferation of HCT116 and HCT15 cells(P＜0.001),induced cell apoptosis(P＜0.001),and led to cell cycle arrest in G1 phase(P＜0.01).RNA-Seq analysis showed that KCMF1 was involved in the regulation of several signaling pathways,including nuclear factor-κB(NF-κB)signaling pathway.KCMF1 knockdown reduced the transcription levels of the target genes of NF-κB signaling pathway,including BCL-XL,XIAP and CIAP(P＜0.05),and suppressed the expression of phosphorylated p65 and nuclear translocation of p65(P＜0.01).Meanwhile,the activity of NF-κB reporter was reduced in tumor cells upon KCMF1 knockdown(P＜0.01).Conclusion:The expression of KCMF1 is significantly upregulated in human CRC tissues and positively associated with advanced clinical stage and poor prognosis.KCMF1 may promote the proliferation of CRC cells by activating the NF-κB signaling pathway.KCMF1 may be a potential new therapeutic target for CRC.

Although immunotherapy has made a breakthrough in the treatment of cancer,the emergence of drug resistance has limited be-nefits in most patients,and reversing immunotherapy resistance remains a hotly debated and unresolved issue.The tumor microenviron-ment(TME)comprises a large number of T lymphocytes.During the biological processes of proliferation,activation,and the effect of tumor-infiltrating lymphocytes(TILs),TILs face severe local metabolic stress,resulting in metabolic reprogramming to meet the markedly increased energy demand and biosynthesis requirements.This process of adaptive changes in the metabolic pattern is closely related to the pheno-type and function of TILs,affecting immunotherapy efficacy and mediating immunotherapy resistance.In recent years,with the study of TIL metabolism and the search for specific metabolic regulatory points,targeting the TIL metabolic pathway could restore the tumor-killing func-tion and reverse immunotherapy resistance.This review explores the relationship between the metabolic reprogramming of TILs and im-mune resistance,and provides a new strategy for reversing immune resistance.

Objective To analyze the effects of arthroscopic meniscus treatment on knee osteoarthritis on the effective rate,VAS score,HSS score and TNF-α level.Methods We selected 86 patients with knee osteoarthritis caused by meniscus injury who were treated in our hospital from June 2019 to May 2020 as the research subjects.The treatment method was selected according to the patient's wishes,with 43 patients who chose conventional conservative treatment included in the control group and 43 patients who chose arthroscopic meniscus therapy included in the study group.The treatment effectiveness,visual ana-logue scale(VAS)score,HSS knee joint score and tumor necrosis factor-α(TNF-α)index levels were compared between the two groups of patients.Results The effective rate of treatment in the study group was significantly higher than that in the control group(P＜0.05);there was no significant difference between the study group and the control group in the VAS score,HSS score,and TNF-α level before treatment(P＞0.05).After treatment,the VAS score,HSS score and TNF-α level of the group were improved.The VAS score and TNF-α level of the study group after treatment were lower than those of the control group(P＜0.05),and the HSS score was higher than the control group.Group(P＜0.05).Conclusion The arthroscopic meniscus treatment for patients with knee osteoarthritis has a significant effect,which can effectively reduce the pain and inflammation of the patients and improve the function of the knee joint.

Objective To investigate the predictive factors for acute kidney injury (AKI) in patients with acute liver failure (ALF), and to establish a new predictive model. Methods Clinical data were collected from 253 patients who were diagnosed with ALF in The First Affiliated Hospital of Zhengzhou University from January 2015 to October 2021, and according to the presence or absence of AKI, these patients were divided into non-AKI group with 170 patients and AKI group with 83 patients. Related clinical data and laboratory markers were collected. Non-normally distributed continuous data were expressed as M ( P 25 - P 75 ), and the Mann-Whitney U test was used for comparison between two groups; categorical data were expressed as cases (%), and the chi-square test was used for comparison between two groups. The binary logistic regression analysis was used to investigate the risk factors for AKI in ALF patients, and the receiver operating characteristic (ROC) curve was used to evaluate the performance of the indices obtained in predicting AKI in ALF patients. Results Compared with the non-AKI group, the AKI group had a significantly higher proportion of patients with hypertension, diabetes, hepatic encephalopathy, ascites, and pulmonary infection, significantly higher levels of white blood cell count (WBC), international normalized ratio (INR), C-reactive protein, procalcitonin (PCT), neutrophil-to-lymphocyte ratio, and Model for End-Stage Liver Disease (MELD) score, and significantly lower levels of platelet count, lymphocyte-to-monocyte ratio, and PNI (all P < 0.05). The multivariate logistic regression analysis showed that WBC (odds ratio [ OR ]=1.267, 95% confidence interval [ CI ]: 1.124-1.428, P < 0.001), INR ( OR =1.663, 95% CI : 1.205-2.293, P =0.002), PCT ( OR =1.416, 95% CI : 1.137-1.764, P =0.002), and MELD score ( OR =1.098, 95% CI : 1.029-1.172, P =0.005) were risk factors for the development of AKI in patients with ALF. The ROC curve analysis showed that the combination of WBC+INR+PCT+MELD had the largest area under the ROC curve (AUC) of 0.908 in predicting AKI in ALF patients, while WBC, INR, PCT, and MELD alone had an AUC of 0.776, 0.771, 0.746, and 0.780, respectively, in predicting AKI. Conclusion WBC, INR, PCT, and MELD score are independent influencing factors for AKI in patients with ALF, and the predictive model established based on these four indices has a relatively high predictive value.

In this study, a single base editing system was used to edit the FecB and GDF9 gene to achieve a targeted site mutation from A to G and from C to T in Ouler Tibetan sheep fibroblasts, and to test its editing efficiency. Firstly, we designed and synthesized sgRNA sequences targeting FecB and GDF9 genes of Ouler Tibetan sheep, followed by connection to epi-ABEmax and epi-BE4max plasmids to construct vectors and electrotransfer into Ouler Tibetan sheep fibroblasts. Finally, Sanger sequencing was performed to identify the target point mutation of FecB and GDF9 genes positive cells. T-A cloning was used to estimate the editing efficiency of the single base editing system. We obtained gRNA targeting FecB and GDF9 genes and constructed the vector aiming at mutating single base of FecB and GDF9 genes in Ouler Tibetan sheep. The editing efficiency for the target site of FecB gene was 39.13%, whereas the editing efficiency for the target sites (G260, G721 and G1184) of GDF9 gene were 10.52%, 26.67% and 8.00%, respectively. Achieving single base mutation in FecB and GDF9 genes may facilitate improving the reproduction traits of Ouler Tibetan sheep with multifetal lambs.
Animals ; Sheep/genetics* ; Gene Editing ; Tibet ; Mutation ; Phenotype ; Mutagenesis, Site-Directed

Objective:To analyze the clinical and histopathological features of patients with chronic hepatitis B (CHB) combined with metabolic-associated fatty liver disease (MAFLD).Methods:Clinical data of 529 cases who had liver biopsies at the First Affiliated Hospital of Zhengzhou University between January 2015 and October 2021 were collected. Among them were 290 cases with CHB, 155 cases with CHB combined with MAFLD, and 84 cases with MAFLD. Three groups of patients clinical data, including general information, biochemical indicators, FibroScan indicators, viral load, and histopathology, were analyzed. A binary logistic regression analysis was used to explore the factors influencing MAFLD in patients with CHB.Results:(1) Age, male status, proportion of hypertension and diabetes, body mass index, fasting blood glucose, γ-glutamyl transpeptidase, low-density lipoprotein, cholesterol, triglycerides, uric acid, creatinine, and the controlled attenuation parameter for hepatic steatosis were higher in CHB combined with MAFLD than in CHB patient groups. In contrast, the high-density lipoprotein, HBeAg positivity rate, viral load level, and liver fibrosis grade (S stage) were lower in CHB patients, and the differences were statistically significant ( P < 0.05). (2) Alanine aminotransferase, aspartate aminotransferase, γ-glutamyl transpeptidase, triglycerides, uric acid, creatinine, and the controlled attenuation parameter for hepatic steatosis in CHB combined with the MAFLD were lower than those in MAFLD patient groups, while high-density lipoprotein was higher than that of MAFLD patients, and the difference was statistically significant ( P < 0.05). There was no statistically significant difference in the grade of liver inflammation and fibrosis (GS stage) between the two groups ( P > 0.05). Binary multivariate logistic regression analysis showed that overweight/obesity, triglycerides, low-density lipoprotein, the controlled attenuation parameter for hepatic steatosis, and HBeAg positivity were independent influencing factors for MAFLD in CHB patients. Conclusion:Patients with CHB combined with metabolic disorders are prone to developing MAFLD, and there is a certain correlation between HBV viral factors, the degree of liver fibrosis, and the fatty degeneration of hepatocytes.

Objective:To compare the differences in the prevalence of mild micro-hepatic encephalopathy (MHE) among patients with cirrhosis by using the psychometric hepatic encephalopathy score (PHES) and the Stroop smartphone application (Encephal App) test.Methods:This prospective, multi-center, real-world study was initiated by the National Clinical Medical Research Center for Infectious Diseases and the Portal Hypertension Alliance and registered with International ClinicalTrials.gov (NCT05140837). 354 cases of cirrhosis were enrolled in 19 hospitals across the country. PHES (including digital connection tests A and B, digital symbol tests, trajectory drawing tests, and serial management tests) and the Stroop test were conducted in all of them. PHES was differentiated using standard diagnostic criteria established by the two studies in China and South Korea. The Stroop test was evaluated based on the criteria of the research and development team. The impact of different diagnostic standards or methods on the incidence of MHE in patients with cirrhosis was analyzed. Data between groups were differentiated using the t-test, Mann-Whitney U test, and χ2 test. A kappa test was used to compare the consistency between groups. Results:After PHES, the prevalence of MHE among 354 cases of cirrhosis was 78.53% and 15.25%, respectively, based on Chinese research standards and Korean research normal value standards. However, the prevalence of MHE was 56.78% based on the Stroop test, and the differences in pairwise comparisons among the three groups were statistically significant (kappa = -0.064, P < 0.001). Stratified analysis revealed that the MHE prevalence in three groups of patients with Child-Pugh classes A, B, and C was 74.14%, 83.33%, and 88.24%, respectively, according to the normal value standards of Chinese researchers, while the MHE prevalence rates in three groups of patients with Child-Pugh classes A, B, and C were 8.29%, 23.53%, and 38.24%, respectively, according to the normal value standards of Korean researchers. Furthermore, the prevalence rates of MHE in the three groups of patients with Child-Pugh grades A, B, and C were 52.68%, 58.82%, and 73.53%, respectively, according to the Stroop test standard. However, among the results of each diagnostic standard, the prevalence of MHE showed an increasing trend with an increasing Child-Pugh grade. Further comparison demonstrated that the scores obtained by the number connection test A and the number symbol test were consistent according to the normal value standards of the two studies in China and South Korea ( Z = -0.982, -1.702; P = 0.326, 0.089), while the other three sub-tests had significant differences ( P < 0.001). Conclusion:The prevalence rate of MHE in the cirrhotic population is high, but the prevalence of MHE obtained by using different diagnostic criteria or methods varies greatly. Therefore, in line with the current changes in demographics and disease spectrum, it is necessary to enroll a larger sample size of a healthy population as a control. Moreover, the establishment of more reliable diagnostic scoring criteria will serve as a basis for obtaining accurate MHE incidence and formulating diagnosis and treatment strategies in cirrhotic populations.

Non-small cell lung cancer(NSCLC) is one of the most common malignancies worldwide. Not only the complex molecular components but also the cellular heterogeneity in NSCLC tissues pose a great barrier for its clinical treatment. Recent years has witnessed the widespread application of single-cell sequencing in the studies regarding tumor heterogeneity. However, the disadvantages of single-cell sequencing technology itself could not be neglected. Spatial transcriptome(ST) technology allows in situ transcriptome sequencing of tissues to achieve high-throughput transcriptomic information of tissue cells with their spatial information available. In other words, ST makes it possible to acquire cellular composition and gene expression patterns without breaking intercellular communication network, which distinguishes itself from conventional single-cell sequencing, since mechanical separation and enzymatic digestion of tissue cells into single-cell suspension used to be inevitable during the performance of single-cell sequencing. To gain new insights into the spatial heterogeneity of NSCLC, we reviewed and summarized the latest progress in ST technology which has been applied to tumor sample analysis, especially to the field of NSCLC.