BACKGROUND: Temozolomide (TMZ) resistance is a significant challenge in treating glioblastoma (GBM). Collagen remodeling has been shown to be a critical factor for therapy resistance in other cancers. This study aimed to investigate the mechanism of TMZ chemoresistance by GBM cells reprogramming collagens. METHODS: Key extracellular matrix components, including collagens, were examined in paired primary and recurrent GBM samples as well as in TMZ-treated spontaneous and grafted GBM murine models. Human GBM cell lines (U251, TS667) and mouse primary GBM cells were used for in vitro studies. RNA-sequencing analysis, chromatin immunoprecipitation, immunoprecipitation-mass spectrometry, and co-immunoprecipitation assays were conducted to explore the mechanisms involved in collagen accumulation. A series of in vitro and in vivo experiments were designed to assess the role of the collagen regulators prolyl 4-hydroxylase subunit alpha 1 (P4HA1) and yes-associated protein (YAP) in sensitizing GBM cells to TMZ. RESULTS: This study revealed that TMZ exposure significantly elevated collagen type I (COL I) expression in both GBM patients and murine models. Collagen accumulation sustained GBM cell survival under TMZ-induced stress, contributing to enhanced TMZ resistance. Mechanistically, P4HA1 directly binded to and hydroxylated YAP, preventing ubiquitination-mediated YAP degradation. Stabilized YAP robustly drove collagen type I alpha 1 ( COL1A1) transcription, leading to increased collagen deposition. Disruption of the P4HA1-YAP axis effectively reduced COL I deposition, sensitized GBM cells to TMZ, and significantly improved mouse survival. CONCLUSION P4HA1 maintained YAP-mediated COL1A1 transcription, leading to collagen accumulation and promoting chemoresistance in GBM.
Temozolomide ; Humans ; Glioblastoma/drug therapy* ; Animals ; Mice ; Cell Line, Tumor ; Drug Resistance, Neoplasm/genetics* ; YAP-Signaling Proteins ; Hydroxylation ; Dacarbazine/pharmacology* ; Adaptor Proteins, Signal Transducing/metabolism* ; Transcription Factors/metabolism* ; Collagen/biosynthesis* ; Collagen Type I/metabolism* ; Prolyl Hydroxylases/metabolism* ; Antineoplastic Agents, Alkylating/therapeutic use*

Objective To investigate the effects of lobaplatin on proliferation and invasion of cervical cancer CaSki cells.Methods Human cervical cancer CaSki cells were randomly divided into blank control group,2,6 and 12 μg/ml lobaplatin groups by random number table method.The proliferations of the cells were detected by methyl thiazolyl tetrazolium (MTT).The morphological changes of the cells were observed by inverted microscope.The invasive abilities of the cells were detected by Transwell invasion test.The protein expressions of extracellular signal-regulated kinase (ERK) and phospho-extracellular signal-regulated kinase (p-ERK) were detected by Western blotting.Results The absorbance (A) values of blank group,2,6,12 μg/ml lobaplatin groups cultured for 24 h were 0.513 ± 0.023,0.428 ± 0.014,0.380 ± 0.012 and 0.300 ± 0.013 respectively,those of the cells cultured for 48 h were 0.831 ± 0.024,0.558 ± 0.019,0.415 ± 0.015 and 0.088 ±0.009 respectively,and those of the cells cultured for 72 h were 1.153 ±0.022,0.572 ± 0.023,0.201 ± 0.017 and 0.052 ± 0.014 respectively.The differences were statistically significant (F =12.922,P ＜ 0.001;F =10.192,P ＜ 0.001;F =11.192,P ＜ 0.001),and the differences between each two groups were statistically significant (all P ＜ 0.05).Under inverted microscope,the cells of the platinum groups were shrunken and round,the volume and quantity were reduced,the morphology was irregular,the gap was increased,and the changes were more obvious with the increase of the concentration and the culture time.The numbers of penetrating cells of the blank group,2,6,12 μg/ml lobaplatin groups were 87.27 ±9.38,71.02 ± 8.92,53.20 ± 10.02 and 21.02 ± 7.37 respectively.The difference was statistically significant (F =87.291,P ＜ 0.001),and the differences between each two groups were statistically significant (all P ＜ 0.05).The A values of ERK protein in the blank group,2,6,12 μ~ml lobaplatin groups (0.955 ± 0.021、0.953 ± 0.023、0.950 ± 0.020、0.951 ±0.022)showed no significant difference (F =2.033,P =0.783),but the A values of p-ERK protein in the four groups were 0.941 ±0.015,0.831 ±0.020,0.620 ±0.019 and 0.493 ±0.017 respectively,which showed significant difference (F =11.921,P ＜0.001),and the differences between each two groups were statistically significant (all P ＜ 0.05).Conclusion Lobaplatin can inhibit the proliferation and invasion of cervical cancer CaSki cells,which may be related to the inhibition of the expression of p-ERK protein.

Objective To evaluate the effect of levamlodipine intervention in diabetic nephropathy patients which accompanied with hypertension, using the technology of diffusion-weighted imaging (DWI) of functional magnetic resonance (fMRI).Methods A controlled prospective method was taken , and fifty diabetic nephropathy ( phase III) patients which accompanied with hypertension were randomized assigned to two groups of A ( n =26) and B ( n =24).Levamlodipine (2.5 mg qd) was taken by patients of group A and amlodipine (5 mg qd) was taken by patients of group B for 24 weeks, respectively.Two groups both took angiotensinⅡreceptor blockers (ARBs) as the first line antithypertensive agents , their urinary albumin excretion rate (UAER), serum creatinine (sCr), cystatin C (Cys C) , and DWI scanning were detected before and after intervention .The levels of UAER, apparent diffusion coeffi-cient (ADC) value were compared between two groups before and after intervention .During the 24th week, two groups'adverse reac-tion to the medicines and the levels of blood pressure were recorded in each follow-up visit.Results The levels of UAER, systolic blood pressure(SBP), and diastolic blood pressure(DBP) were Significantly lower in group A after 24-week intervention compared to baseline [42.5 (25.3~91.0)μg/min vs 49.2(29.7~96.8)μg/min,(112.6 ±6.4)mmHg vs (135.3 ±7.6)mmHg, (71.4 ± 10.7)mmHg vs (80.3 ±11.6)mmHg, P <0.05, respectively].DWI scanning showed that ADC value of renal parenchyma was sig-nificantly improved than that of baseline [(2.45 ±0.12)vs(2.17 ±0.09), P <0.05].In Group B, the level of SBP was also signifi-cantly lower than that of baseline [(121.5 ±11.6)mmHg vs (134.8 ±9.2)mmHg, P <0.05], and ADC value of renal parenchyma was significantly improved than that of baseline [(2.28 ±0.15) vs (2.14 ±0.09), P <0.05].No difference was found in DBP and UAER before and after intervention ( P >0.05).Group A had a better improvement of SBP (ΔSBP) and ADC (ΔADC) after inter-vention compared to group B ( P =0.02,0.01, respectively).The overall adverse reaction incidence was 15.4%(4/26) in group A and 41.7%(10/24)in group B, respectively (χ2 =4.27, P =0.0387).Conclusions For the diabetic nephropathy (phase III) pa-tients accompanied with hypertension , levamlodipine likely showed better effects on reducing comprehensive blood pressure and UAER , improving renal microcirculation , with less overall adverse reaction compared to amlodipine .

Objective To explore the expression level and clinical significance of multifunctional CD8 T cells in patients with tuberculosis (TB). Methods The expression levels of MTB antigen specific and non-specific multifunctional CD8 T cells among peripheral blood mononuclear cells (PBMCs) and pleural fluid mononuclear cells (PFMCs) in TB patients, latent tuberculosis infection patients (LTBI) and healthy controls (HC) were measured by flow cytometry. Results The expression level of multifunctional CD8 T cells (IL-2+IFN-γ+TNF-α+CD8 T cells) among PBMCs stimulated by non-specific MTB antigen in TB patients was (5.72 ± 4.32)%, which was significantly lower than those in HC and LTBI [(22.3 ± 15.7)%, q=7.455, P<0.001;(14.2 ± 7.72)%, q=3.110, P<0.05]. Under the stimulation by specific MTB antigen, the expression level of multifunctional CD8 T cells among PBMCs in TB patients was (0.33 ± 0.83)%, which was significantly higher than those in HC and LTBI [(0.017 ± 0.03)%, q=3.97, P<0.05;(0.019 ± 0.035)%, q=3.39, P<0.05]. In patients with tuberculous pleurisy, the expression level of multifunctional CD8 T cells among PFMCs was (0.623 ± 1.033)%, which was significantly higher than that among PBMCs [(0.034 ± 0.066)%, P<0.001]. The expression level of multifunctional CD8 T cells in TB patients was negatively correlated with HRCT score (r=-0.265 8, P=0.015 8). Conclusion The expression level of multifunctional CD8 T cells was contributed to discriminate TB patients from latent tuberculosis infection patients , and was closely related to the degree of damage in lung.

Objective To observe the effect of JWSNS serum on proliferation and apoptosis hepatic stellate cells.Methods After being added different concentrations of JWSNS (the low concentrations of JWSNS:0.78 g/ml of crude drug; the medium concentration group of JWSNS:1.56 g/ml of crude drug; the high concentration group of JWSNS:3.12 g/ml of crude drug) drug-containing serum in vitro HSC-T6 cells for 12h,24 h and 48 h respectively,detected serum HSC-T6 proliferation with MTT colorimetry method and measured HSC-T6 apoptosis with flow cytometry and TUNEL method.Results (①) After applied JWSNS on rats HSC-T6,the Cell proliferation was inhibited which showed a time-concentration dependence.The differences were significant when comparing each JWSNS group with the control group (P＜0.01).High concentration of JWSNS group showed significant difference when compared with Biejiaruangan tablets group (P＜0.05) with high concentration of JWSNS (0.399± 0.041) ％ after 48h,and Biejia-Ruangan tablets (0.429± 0.037) ％ after 48 h.② Flow cytometry analysis showed each JWSNS group and Biejiaruangan tablets group had significant increased cell apoptosis when compared with the control group (P＜0.05) after 12 h,24 h,and 48 h.JWSNS medium concentration group [12 h was (17.83±0.25)％,24 h was (26.06±0.26)％,48 h was (39.30±2.25) ％] and JWSNS high concentration group [12 h was (27.15±0.29)％,24 h was (38.96±0.51)％,48 h was (49.34± 0.77) ％] had a significant increased cell apoptosis compared to the Biejia-Ruangan tablets group [ 12 h was (8.31 ± 0.30) ％,24 h was (16.25 ± 0.25) ％,48 h was (27.12± 0.39) ％].③ TUNEL detection showed that each concentration of JWSNS group [the low concentration of JWSNS:was (25.1 ± 1.48)％,medium concentration group of JWSNS was(39.30±2.25)％,high concentration group of JWSNS was(39.30±2.25)％] had a significant increased cell apoptosis rate than Biejiaruangan tablets group (30.0± 3.92) after 48 h (P＜0.01).Conclusion JWSNS containing serum can inhibit the proliferation of HSC-T6 in vitro,promote the apoptosis

The aim of this study was to investigate the effect of Paris saponin I (PS I) on human gastric carcinoma cell growth and apoptosis and to explore the potential mechanisms. The proliferation of SGC7901 cells was monitored by the MTT cell viability assay, while the nuclear morphology of apoptotic cells was assessed by Hoechst 33258 staining. Flow cytometry was performed to analyze the cell cycle progression of propidium iodide (PI)-stained SGC7901 cells and the apoptotic rate of annexin V/PI-stained cells. Western blotting was used to examine the expression of several cell cycle proteins, including cyclin B1 and Cdk1, and the apoptosis-regulated proteins Bcl-2, Bax, cytochrome c, procaspase-9, and procaspase-3. The MTT assay demonstrated that PS I could induce significant dose- and time-dependent inhibition of SGC7901 cell proliferation. Marked morphological changes, including condensation of chromatin, nuclear fragmentation and apoptotic bodies were clearly shown on Hoechst 33258 staining. PSI treatment also resulted in the disruption of the cell cycle at G(2)/M and the induction of apoptosis. Following PSI treatment, the cell cycle-related proteins cyclin B1 and Cdk1 were down-regulated. Expression of the pro-apoptotic protein Bax was increased, while anti-apoptotic protein Bcl-2 decreased. PSI treatment resulted in elevated cytoplasmic cytochrome c and activation of the apoptotic proteases caspase-9 and caspase-3. These data indicate that PS acts as an inhibitor of proli I feration in SGC7901 cells by inducing cell cycle arrest and mitochondria-dependent apoptosis. PSI is a potential therapeutic agent against human gastric carcinoma.

The aim of this study was to investigate the effect of Paris saponin Ⅰ (PS Ⅰ ) on human gastric carcinoma cell growth and apoptosis and to explore the potential mechanisms.The proliferation of SGC7901 cells was monitored by the MTT cell viability assay,while the nuclear morphology of apoptotic cells was assessed by Hoechst 33258 staining.Flow cytometry was performed to analyze the cell cycle progression of propidium iodide (PI)-stained SGC7901 cells and the apoptotic rate of annexin V/PI-stained cells.Western blotting was used to examine the expression of several cell cycle proteins,including cyclin B1 and Cdkl,and the apoptosis-regulated proteins Bcl-2,Bax,cytochrome c,procaspase-9,and procaspase-3.The MTT assay demonstrated that PSⅠ could induce significant doseand time-dependent inhibition of SGC7901 cell proliferation.Marked morphological changes,including condensation of chromatin,nuclear fragmentation and apoptotic bodies were clearly shown on Hoechst 33258 staining.PSⅠ treatment also resulted in the disruption of the cell cycle at G2/M and the induction of apoptosis.Following PSⅠ treatment,the cell cycle-related proteins cyclin B 1 and Cdk1 were downregulated.Expression of the pro-apoptotic protein Bax was increased,while anti-apoptotic protein Bcl-2decreased.PSⅠ treatment resulted in elevated cytoplasmic cytochrome c and activation of the apoptotic proteases caspase-9 and caspase-3.These data indicate that PSⅠ acts as an inhibitor of proliferation in SGC7901 cells by inducing cell cycle arrest and mitochondria-dependent apoptosis.PSⅠ is a potential therapeutic agent against human gastric carcinoma.

Adult ; Blindness ; etiology ; Cysts ; surgery ; Humans ; Infection ; complications ; etiology ; Male ; Paranasal Sinus Diseases ; surgery ; Postoperative Complications ; Sphenoid Sinus