Objective To analyze the feasibility of probiotics in delaying the progression of early Alzheimer's disease(AD)based on the changes of brain-gut-microbiota axis factors.Methods A total of 100 patients with early AD who visited Shaoxing 7th People's Hospital from December 2023 to December 2024 were selected and divided into control group(40 cases)and study group(60 cases)based on different treatment methods.The control group was treated with donepezil hydrochloride,while study group was treated with donepezil hydrochloride combined with probiotic preparations.The cognitive function,daily living ability,intestinal flora,and relative expression levels of brain-gut-microbiota axis factors,interleukin(IL)-6,tumor necrosis factor-α(TNF-α),5-hydroxytryptamine(5-HT),γ-aminobutyric acid(GABA),and lipopolysaccharide(LPS)/Toll-like receptor 4(TLR4)/nuclear factor-KB(NF-κB)axis factors were compared before and after treatment in both groups.The clinical efficacy and prognosis monitoring results were also compared.Results The total effective rate and the proportion of patients with a clinical dementia rating score of 0 in study group were higher than those in control group(P＜0.05).The cognitive function and daily living ability scores in study group were higher than those in control group(P＜0.05).The copy numbers of lactobacillus and bifidobacterium in study group were higher than those in control group,while the copy numbers of enterococcus and escherichia coli were lower(P＜0.05).The relative expression levels of amyloid-β protein 1-42,IL-6,TNF-α,TLR4,and NF-κB in study group were lower than those in control group,and the levels of 5-HT and GABA were higher(P＜0.05).Conclusion Probiotics can delay the progression of early AD patients and improve clinical efficacy.It is speculated that the mechanism of action may be related to regulating the brain-gut-microbiota axis-mediated LPS/TLR4/NF-κB pathway to restore the intestinal microecology and alleviate neurological inflammation.

Objective To analyze the feasibility of probiotics in delaying the progression of early Alzheimer's disease(AD)based on the changes of brain-gut-microbiota axis factors.Methods A total of 100 patients with early AD who visited Shaoxing 7th People's Hospital from December 2023 to December 2024 were selected and divided into control group(40 cases)and study group(60 cases)based on different treatment methods.The control group was treated with donepezil hydrochloride,while study group was treated with donepezil hydrochloride combined with probiotic preparations.The cognitive function,daily living ability,intestinal flora,and relative expression levels of brain-gut-microbiota axis factors,interleukin(IL)-6,tumor necrosis factor-α(TNF-α),5-hydroxytryptamine(5-HT),γ-aminobutyric acid(GABA),and lipopolysaccharide(LPS)/Toll-like receptor 4(TLR4)/nuclear factor-KB(NF-κB)axis factors were compared before and after treatment in both groups.The clinical efficacy and prognosis monitoring results were also compared.Results The total effective rate and the proportion of patients with a clinical dementia rating score of 0 in study group were higher than those in control group(P＜0.05).The cognitive function and daily living ability scores in study group were higher than those in control group(P＜0.05).The copy numbers of lactobacillus and bifidobacterium in study group were higher than those in control group,while the copy numbers of enterococcus and escherichia coli were lower(P＜0.05).The relative expression levels of amyloid-β protein 1-42,IL-6,TNF-α,TLR4,and NF-κB in study group were lower than those in control group,and the levels of 5-HT and GABA were higher(P＜0.05).Conclusion Probiotics can delay the progression of early AD patients and improve clinical efficacy.It is speculated that the mechanism of action may be related to regulating the brain-gut-microbiota axis-mediated LPS/TLR4/NF-κB pathway to restore the intestinal microecology and alleviate neurological inflammation.

Objective:To explore the role and molecular mechanism of hepatocyte nuclear factor 4γ (HNF4γ) in proliferation and stemness of gastric cancer.Methods:A total of 102 cases of paraffin-embedded gastric cancer tissues and matched adjacent gastric tissues and 42 cases of fresh-frozen tissues derived from gastric patients who received radical gastrectomy were collected from the First Affiliated Hospital of Zhengzhou University between 2012 to 2015. The expression of HNF4γ was tested by immunohistochemical staining, quantitative real-time polymerase chain reaction (qRT-PCR). HNF4γ overexpressed (AGS-HNF4γ) and shRNA silenced (HGC27-shHNF4γ) gastric cell lines were established. The effects of HNF4γ on cell proliferation and stemness were verified by XTT, clone formation and sphere formation assay. The expression of CD44 was detected by western blot.Results:The mRNA expression level of HNF4γ in fresh-frozen gastric cancer tissue was (12.43±2.702), which was significantly higher than (3.639±1.109) in normal tissue ( P<0.001). The high protein expression rate of HNF4γ in paraffin-embedded gastric cancer tissues was 41.2% (42/102), which was significantly higher than 8.8% (9/102) in normal gastric mucosa tissue ( P< 0.001). The protein expression of HNF4γ was closely related to the tumor differentiation, infiltration depth, lymph node metastasis and tumor stage ( P<0.05). The median survival interval of patients with HNF4γ high expression was 25 months, the 3-year survival rate was 4.8% (2/42), significantly lower than 38 months and 51.7% (31/60) of patients with normal HNF4γ expression ( P<0.001). The proliferation and CD44 protein expression of AGS-HNF4γ cells were significantly higher than those of the AGS-Vector cells. The number of clone formation, sphere formation rate of AGS-HNF4γ cells were 243.5±24.5 and (83.5±3.9)%, significantly higher than 81.0±16.0 and (21.8±5.6)% of AGS-Vector cells ( P=0.030 and P=0.010, respectively). The proliferation and CD44 protein expression of HGC27-shHNF4 cells were significantly lower than those of the HGC27-vector cells. The number of clone formation, sphere formation rate of HGC27-shHNF4 cells were 26.0±1.0 and (20.8±8.4)%, significantly higher than 83.5±4.5 and (72.5±4.8)% of HGC27-vector cells ( P=0.006 and P=0.030, respectively). Conclusions:HNF4γ is upregulated in the gastric cancer tissues and related with the poor prognosis of patients with gastric cancer. Overexpression of HNF4γ promotes the proliferation and remains the stemness of gastric cancer cells by upregulating the expression of CD44.

Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and is the fourth-leading cause of cancer-related deaths worldwide. HCC is refractory to many standard cancer treatments and the prognosis is often poor, highlighting a pressing need to identify biomarkers of aggressiveness and potential targets for future treatments. Kinesin family member 2C (KIF2C) is reported to be highly expressed in several human tumors. Nevertheless, the molecular mechanisms underlying the role of KIF2C in tumor development and progression have not been investigated. In this study, we found that KIF2C expression was significantly upregulated in HCC, and that KIF2C up-regulation was associated with a poor prognosis. Utilizing both gain and loss of function assays, we showed that KIF2C promoted HCC cell proliferation, migration, invasion, and metastasis both in vitro and in vivo. Mechanistically, we identified TBC1D7 as a binding partner of KIF2C, and this interaction disrupts the formation of the TSC complex, resulting in the enhancement of mammalian target of rapamycin complex1 (mTORC1) signal transduction. Additionally, we found that KIF2C is a direct target of the Wnt/β-catenin pathway, and acts as a key factor in mediating the crosstalk between Wnt/β-catenin and mTORC1 signaling. Thus, the results of our study establish a link between Wnt/β-catenin and mTORC1 signaling, which highlights the potential of KIF2C as a therapeutic target for the treatment of HCC.
Adult ; Aged ; Animals ; Carcinoma, Hepatocellular/pathology* ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Epithelial-Mesenchymal Transition/genetics* ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Intracellular Signaling Peptides and Proteins/metabolism* ; Kinesins/metabolism* ; Liver Neoplasms/pathology* ; Male ; Mice ; Mice, Inbred BALB C ; Middle Aged ; Neoplasm Staging ; Prognosis ; Protein Binding ; RNA, Small Interfering/metabolism* ; Survival Analysis ; Tumor Burden ; Wnt Signaling Pathway ; Xenograft Model Antitumor Assays ; beta Catenin/metabolism*

Objective:To explore the role and molecular mechanism of hepatocyte nuclear factor 4γ (HNF4γ) in proliferation and stemness of gastric cancer.Methods:A total of 102 cases of paraffin-embedded gastric cancer tissues and matched adjacent gastric tissues and 42 cases of fresh-frozen tissues derived from gastric patients who received radical gastrectomy were collected from the First Affiliated Hospital of Zhengzhou University between 2012 to 2015. The expression of HNF4γ was tested by immunohistochemical staining, quantitative real-time polymerase chain reaction (qRT-PCR). HNF4γ overexpressed (AGS-HNF4γ) and shRNA silenced (HGC27-shHNF4γ) gastric cell lines were established. The effects of HNF4γ on cell proliferation and stemness were verified by XTT, clone formation and sphere formation assay. The expression of CD44 was detected by western blot.Results:The mRNA expression level of HNF4γ in fresh-frozen gastric cancer tissue was (12.43±2.702), which was significantly higher than (3.639±1.109) in normal tissue ( P<0.001). The high protein expression rate of HNF4γ in paraffin-embedded gastric cancer tissues was 41.2% (42/102), which was significantly higher than 8.8% (9/102) in normal gastric mucosa tissue ( P< 0.001). The protein expression of HNF4γ was closely related to the tumor differentiation, infiltration depth, lymph node metastasis and tumor stage ( P<0.05). The median survival interval of patients with HNF4γ high expression was 25 months, the 3-year survival rate was 4.8% (2/42), significantly lower than 38 months and 51.7% (31/60) of patients with normal HNF4γ expression ( P<0.001). The proliferation and CD44 protein expression of AGS-HNF4γ cells were significantly higher than those of the AGS-Vector cells. The number of clone formation, sphere formation rate of AGS-HNF4γ cells were 243.5±24.5 and (83.5±3.9)%, significantly higher than 81.0±16.0 and (21.8±5.6)% of AGS-Vector cells ( P=0.030 and P=0.010, respectively). The proliferation and CD44 protein expression of HGC27-shHNF4 cells were significantly lower than those of the HGC27-vector cells. The number of clone formation, sphere formation rate of HGC27-shHNF4 cells were 26.0±1.0 and (20.8±8.4)%, significantly higher than 83.5±4.5 and (72.5±4.8)% of HGC27-vector cells ( P=0.006 and P=0.030, respectively). Conclusions:HNF4γ is upregulated in the gastric cancer tissues and related with the poor prognosis of patients with gastric cancer. Overexpression of HNF4γ promotes the proliferation and remains the stemness of gastric cancer cells by upregulating the expression of CD44.

Objective: To assess the vaccine loss related to the Expanded Program on Immunization (EPI) in Xinjiang Uygur Autonomous Region so as to improve the management of vaccines. Methods: A total of 135 vaccination clinics were randomly selected, using a stratified cluster sampling method. In each clinic, data on vaccination was collected between 2016 and 2017, including the number of doses in routine immunization program and supplementary immunization activities (i.e., vaccine doses in vials that were opened for use) on polio vaccine, number of doses administered to children and the number of doses discarded (e.g., expired vaccine or broken vials that had not been opened for use), etc. Coefficient on vaccine loss was calculated with the following equation: vaccine loss coefficient=(number of vaccine doses used)/(number of vaccine doses administered). The vaccine discard rate appeared as: number of vaccine doses discarded)/number of vaccine doses used. Results: For vaccines in single-dose vials [diphtheria-tetanus-pertussis vaccine (DTaP) and trivalent oral polio virus vaccine (tOPV)], the loss coefficients appeared as 1.00 and 1.02, respectively. For vaccines in multi-dose vials [bivalent oral polio vaccine (bOPV), group A meningococcal polysaccharide vaccine (MPV-A), diphtheria-tetanus combined vaccine (DT) and bacilli Calmette-Guérin (BCG) vaccine], the loss coefficients were 1.58, 1.67, 1.68, and 3.02, respectively. The coefficients of EPI vaccine loss in urban, rural, and pastoral area vaccination clinics ranged between 1.00-2.84, 1.00-3.71, and 1.00-2.27, respectively. Loss coefficients ranged between 1.00-3.00, 1.00- 4.41, and 1.00-1.94, respectively, were seen in township clinics, village clinics, and decentralized vaccination clinics. Coefficients on larger vaccine loss were associated with longer intervals between clinic sessions and with fewer vaccinations administrations per day. Conclusions In Xinjiang, coefficients on the loss of multi-dose EPI vaccines were high. The coefficients on loss were different from the levels of region and types of clinics, and time interval between clinic sessions. Programs on refining the management and distribution of EPI vaccines, to minimize the vaccine loss were recommended.

Objective: To investigate the expression of ALC1 protein during esophageal squamous cell carcinoma (ESCC) development and progression, so as to explore its association with clinicopathological characteristics and overall survival of ESCC patients, and the effect of ALC1 overexpression on malignant biological behavior of esophageal squamous cells. Methods: Immunohistochemistry (IHC) was used to detect ALC1 protein expression in 245 primary ESCC tissues and their paired normal esophageal mucous membranes, and to determine its correlation to gender, age, tumor cell differentiation, invasion, TNM stage, lymph nodes metastasis, and overall surviv-al rate of ESCC patients. MTT assay, colony formation assay, transwell invasion, and wound healing assay were used to observe the ef-fect of ALC1 on ESCC cell proliferation, invasion, and migration. Results: The expression ratio of ALC1 in esophageal squamous cell car-cinoma was higher compared with that in their paired normal esophageal mucous membranes (41.6% vs . 21.2% , P<0.05). Upregula-tion of ALC1 was associated with ESCC invasion, TNM stage, and lymph node metastasis (P<0.05). The overall survival of ESCC patients with ALC1 overexpression was significantly lower than that in patients with downregulated ALC1 expression (P=0.002). Therefore, ALC1 may promote the proliferation, invasion, and migration of ESCC cells. Conclusions: ALC1 upregulation may play an important role in the progression and development of ESCC. Upregulation of ALC1 leads to poorer disease prognosis, and could promote the prolifera-tion, invasion, and migration of the KYSE30 ESCC cells. Therefore, ALC1 may have potential prognostic value for ESCC patients.

Objective To investigate the role of cell adhesion molecule L1 like (CALL) in the genesis and development of esophageal squamous cell carcinoma (ESCC).Methods From July 2007 to December 2010,a total of 100 patients with ESCC who received radical resection of esophageal cancer were enrolled.The ESCC tissues and corresponding tumor-adjacent normal tissues were obtained.The expression of CALl was determined by tissue microarray technology and immunohistochemical staining.The CALL over-expressed esophageal cancer cell line was established.The effects of CALL on cell migration and invasion were detected by wound-healing assay and Transwell assay,respectively.The effects of CALL on actin microfilament was analyzed by filamentous actin (F-actin) staining.Chi square test,Fisher's exact test,multivariate analysis and t test were performed for statistical analysis.Results The positive expression rate of CALL in ESCC tissues was 56 ％ (56/100),which was lower than that of tumor-adjacent normal tissues (95％,95/100),and the difference was statistically significant (x2=41.114,P＜0.01).There were statistically significant differences in CALL expression at protein level among patients with ESCC of different differentiation degree,different pathological T stage,lymph node metastasis and different TNM stage (x2=13.702,5.317,21.453,Fisher's exact test;all P＜ 0.05).The five year disease related survival rate of ESCC patients with down-regulated expression of CALL was 0(0/49),which was lower than those with normal CALL expression (25.5％,13/51),and the difference was statistically significant (x2 =43.338,P＜0.01).The median survival time of CALL expression down-regulated group was 17 months,and that of normal expressed group was 38 months.CALL expression was an independent risk factor of disease special survival rate (hazard ratio (HR) 0.353,95％ confidence interval (CI) 0.188 to 0.666,P=0.001).The results of wound-healing assay showed that the migration ability of CALL overexpressed CALL-k30 cells was lower than that of Vec-k30 cells in control group on 24 hours after wound.The results of Transwell invasion test showed the number of migrating cells penetrating CALL k30 cells attached to the inferior surface of the membrane was 44.000±13.748,which was less than that of the Vec k30 cells (154.333±25.007),and the difference was statistically significant (t=5.136,P=0.036).The results of F-actin staining demonstrated that actin filaments of CALL-k30 cells was 234.667 ± 65.118,which was lower than that of Vec-k30 cells (597.000± 119.929),and the difference was statistically significant (t=4.707,P=0.042).Conclusions CALL lowers the migration and invasion abilities of esophageal cancer cells by inhibiting F-actin microfilaments.Its abnormal expression may play an important role in the genesis,development and prognosis of ESCC.

The event-related potential (ERP) is considered as one of the most effective methods to study and analyze objectively human mental activity based on nerve electrophysiology. At present, ERP is not only used in the study of lie detection, but also in the clinical medicine for the cognitive assess-ment on patients with cerebrovascular disease, dementia or traumatic brain injury and auxiliary diagnosis of mental illness. with the further development of ERP detection technology, it would have a wider ap-plication prospect in the field of forensic medicine.

OBJECTIVETo investigate the role of type I collagen as an autocrine protein in promoting the growth lung cancer cells in a three-dimensional (3D) culture system.

METHODSImmunohistochemistry and RT-PCR were used to detect the expression of type I collagen in lung cancer specimens and 5 lung cancer cell lines. The lung cancer cell lines in 3D cultures were treated with vectors harboring short hairpin RNA (shRNA) targeting type I collagen, and the cell growth suppression was evaluated using MTT assay and colony formation assay. The lung cancer cells stimulated with supernatant from lung cancer-derived fibroblasts were tested for the expression of type I collagen mRNA.

RESULTSType I collagen expressions were detected in both the lung cancer tissues and the cell lines. In the 3D culture system, the growth of the cancer cell lines was obviously suppressed by shRNA-mediated type I collagen knockdown evidenced by lowered cell growth rate and colony formation ability. Stimulation with fibroblast culture supernatant resulted in enhanced expressions of type I collagen in the cancer cell lines.

CONCLUSIONAutocrine of type I collagen I is required for maintaining lung cancer cell growth in 3D cultures, and its expression is regulated by fibroblasts. These findings provide new insights into the role of type I collagen I in the tumor microenvironment and point a new direction for targeted therapy of tumors.

Cell Cycle ; Cell Line, Tumor ; Cell Proliferation ; Collagen Type I ; secretion ; Fibroblasts ; Humans ; Immunohistochemistry ; Lung Neoplasms ; metabolism ; RNA, Messenger ; RNA, Small Interfering