Objective:To investigate the impact of sarcopenia on the short-term outcomes and long-term prognosis in cervical cancer patients undergoing concurrent chemoradiotherapy (CCRT).Methods:A total of 410 cervical cancer patients who received CCRT in Henan Provincial People′s Hospital between January 2017 and December 2021 were prospectively enrolled in this study. They were divided into the sarcopenia and non-sarcopenia groups based on the body muscle content measured using bioelectrical impedance analysis. Short-term outcomes were assessed according to the Response Evaluation Criteria in Solid Tumors (RECIST), and acute adverse reactions were assessed based on the toxicity criteria of the Radiation Therapy Oncology Group (RTOG). CCRT termination or prolonged treatment associated with various acute adverse reactions were recorded. All patients were followed up with overall survival (OS) and progression-free survival (PFS) as endpoints. Finally, the survival rate was estimated and the association between sarcopenia and PFS was analyzed.Results:Among the patients, 152 (37.1%) had sarcopenia. Compared to the non-sarcopenia group, the sarcopenia group exhibited higher incidences of grade 2 or above acute adverse reactions in the lower gastrointestinal and hematological systems, CCRT termination, or prolonged treatment. In the non-sarcopenia group, 27 deaths were recorded, with an OS of 30 (18-36) months, a 3-year OS rate of 88.7%, and a 5-year OS rate of 85.6%. In the sarcopenia group, 23 deaths were found, with an OS of 24 (15-33) months, a 3-year OS rate of 83.8%, and a 5-year OS rate of 77.7%. There was no significant difference in survival curves between both groups ( P > 0.05). In the non-sarcopenia group, 52 cases of recurrence were recorded, with a PFS of 21 (12-33) months, a 3-year PFS rate of 77.9%, and a 5-year PFS rate of 71.0%. In the sarcopenia group, 41 cases of recurrence were found, with a PFS of 15 (10.5-24) months, a 3-year PFS rate of 69.0%, and a 5-year PFS rate of 56.5%. There was a significant difference in the PFS curves between both groups ( χ2 = 5.89, P = 0.015). Multivariate Cox regression analysis identified sarcopenia as an independent risk factor for PFS ( χ2 =4.33, P = 0.037). Conclusions:Sarcopenia increases the risks of acute adverse reactions and long-term recurrence in cervical cancer patients undergoing CCRT.

With various diseases ravaging internationally, the demands for recombinant adenoviral vector (Adv) vaccines have increased dramatically. To meet the demand for Adv vaccine, development of a new cell culture process is an effective strategy. Applying hyperosmotic stress in cells before virus infection could increase the yield of Adv in batch culture mode. Emerging perfusion culture can significantly increase the yield of Adv as well. Therefore, combining the hyperosmotic stress process with perfusion culture is expected to improve the yield of Adv at high cell density. In this study, a shake flask combined with a semi-perfusion culture was used as a scaled-down model for bioreactor perfusion culture. Media with osmotic pressure ranging from 300 to 405 mOsm were used to study the effect of hyperosmotic stress on cell growth and Adv production. The results showed that using a perfusion culture process with a hyperosmotic pressure medium (370 mOsm) during the cell growth phase and an isosmotic pressure medium (300 mOsm) during the virus production phase effectively increased the yield of Adv. This might be due to the increased expression of HSP70 protein during the late phases of virus replication. The Adv titer in a bioreactor with such a process reached 3.2×10¹⁰ IFU/mL, three times higher than that of the traditional perfusion culture process. More importantly, this is the first time that a strategy of combining the hyperosmotic stress process with perfusion culture is applied to the production of Adv in HEK 293 cells. It also reveals the reason why the hyperosmotic stress process increased the yield of Adv, which may facilitate the process optimization of for producing other Adv in HEK 293 cells.
Humans ; HEK293 Cells ; Genetic Vectors/genetics* ; Batch Cell Culture Techniques ; Bioreactors ; Perfusion

Objective:To explore the role and molecular mechanism of trophoblastic cell surface antigen 2 (Trop2) in the invasion and migration of ovarian cancer.Methods:Through the data mining of Cancer Cell Line Encyclopedia and TCGA database, the clinical significance of Trop2 expression was analyzed. Western blot was used to detect Trop2 protein expression in ovarian cancer cell lines including A3O, A1780 and SKOV3. SKOV3 cells were used to construct Trop2-short hairpin RNA (shRNA) cell model. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the SKOV3 mRNA expression in SKOV3-shRNA and SKOV3-NC cells. Cell counting kit-8 (CCK8) was used to detect the proliferation of SKOV3-shRNA cells and SKOV3-NC cells. Flow cytometry was used to detect cell cycle and apoptosis in two groups of cells. Transwell array was used to detecte the invasion and migration of SKOV3-shRNA cells and SKOV3-NC cells. Western blot was used to detect the protein expressions of AKT, p-AKT, β-catenin, caspase3, bcl-2, E-cadherin and vimentin.Results:Trop2 mRNA highly expressed in ovarian cancer, and was related to the tumor stage and patient survival. Compared with A3O cells, Trop2 overexpressed in A1780 and SKOV3 cells ( P<0.05). The relative expression levels of Trop2 mRNA in SKOV3-NC group and SKOV3-shRNA group were 1.18±0.24 and 0.42±0.08, with statistically significant difference ( P<0.05). The results of CCK-8 array showed that the cell viability of SKOV3-NC group was significantly higher than that of SKOV3-shRNA group ( P<0.05). The proportion of G 0/G 1 cells in SKOV3-NC and SKOV3-shRNA groups were (38.67±4.22)% and (60.24±8.17)%, respectively. G 0/G 1 arrest was observed in SKOV3-shRNA cells ( P<0.05). The apoptosis rate of SKOV3-shRNA group was (26.32±1.81)%, significantly higher than (6.54±1.32)% of SKOV3-NC group ( P<0.05). The number of migrating SKOV3 cells in the SKOV3-shRNA and SkOV3-NC groups were 1 255.83±108.44 and 1 679.71±213.92, while the number of invading cells were 242.49±52.09 and 473.54±73.11, respectively. Compared with the SKOV3-NC group, the number of migrating and invading SKOV3-shRNA group was significantly reduced (all P<0.05). The expressions of p-AKT2, Bcl-2, vimentin and β-catenin were down-regulated, and the expressions of caspase 3 and E-cadherin were up-regulated in SKOV3-shRNA cells. There was no significant change in the total protein level of AKT. Conclusions:Trop2 expression is related to ovarian cancer stage and postoperative survival. Trop2 can promote ovarian cancer cell proliferation and metastasis by activating the AKT/β-catenin signaling pathway and knockdown of Trop2 inhibits the progression of ovarian cancer.

Objective:To explore the role and molecular mechanism of trophoblastic cell surface antigen 2 (Trop2) in the invasion and migration of ovarian cancer.Methods:Through the data mining of Cancer Cell Line Encyclopedia and TCGA database, the clinical significance of Trop2 expression was analyzed. Western blot was used to detect Trop2 protein expression in ovarian cancer cell lines including A3O, A1780 and SKOV3. SKOV3 cells were used to construct Trop2-short hairpin RNA (shRNA) cell model. Quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) was used to detect the SKOV3 mRNA expression in SKOV3-shRNA and SKOV3-NC cells. Cell counting kit-8 (CCK8) was used to detect the proliferation of SKOV3-shRNA cells and SKOV3-NC cells. Flow cytometry was used to detect cell cycle and apoptosis in two groups of cells. Transwell array was used to detecte the invasion and migration of SKOV3-shRNA cells and SKOV3-NC cells. Western blot was used to detect the protein expressions of AKT, p-AKT, β-catenin, caspase3, bcl-2, E-cadherin and vimentin.Results:Trop2 mRNA highly expressed in ovarian cancer, and was related to the tumor stage and patient survival. Compared with A3O cells, Trop2 overexpressed in A1780 and SKOV3 cells ( P<0.05). The relative expression levels of Trop2 mRNA in SKOV3-NC group and SKOV3-shRNA group were 1.18±0.24 and 0.42±0.08, with statistically significant difference ( P<0.05). The results of CCK-8 array showed that the cell viability of SKOV3-NC group was significantly higher than that of SKOV3-shRNA group ( P<0.05). The proportion of G 0/G 1 cells in SKOV3-NC and SKOV3-shRNA groups were (38.67±4.22)% and (60.24±8.17)%, respectively. G 0/G 1 arrest was observed in SKOV3-shRNA cells ( P<0.05). The apoptosis rate of SKOV3-shRNA group was (26.32±1.81)%, significantly higher than (6.54±1.32)% of SKOV3-NC group ( P<0.05). The number of migrating SKOV3 cells in the SKOV3-shRNA and SkOV3-NC groups were 1 255.83±108.44 and 1 679.71±213.92, while the number of invading cells were 242.49±52.09 and 473.54±73.11, respectively. Compared with the SKOV3-NC group, the number of migrating and invading SKOV3-shRNA group was significantly reduced (all P<0.05). The expressions of p-AKT2, Bcl-2, vimentin and β-catenin were down-regulated, and the expressions of caspase 3 and E-cadherin were up-regulated in SKOV3-shRNA cells. There was no significant change in the total protein level of AKT. Conclusions:Trop2 expression is related to ovarian cancer stage and postoperative survival. Trop2 can promote ovarian cancer cell proliferation and metastasis by activating the AKT/β-catenin signaling pathway and knockdown of Trop2 inhibits the progression of ovarian cancer.

Objective To establish a non-venous bypass orthotopic liver transplantation model in Bama miniature pigs with high repeatability and stability. Methods Twelve Bama miniature pigs were randomly divided into the donor group (n=6) and recipient group (n=6). Pigs underwent non-venous bypass orthotopic liver transplantation. The time of anhepatic phase during operation was shortened, blood pressure during anhepatic phase was stably maintained, and management of anesthesia and body fluid during operation were strengthened. The operation time, anhepatic phase and survival status of the recipients were observed and recorded. The intraoperative heart rate, mean arterial pressure (MAP) and changes in arterial blood gas analysis were monitored. The perioperative liver function was evaluated. Results Among 6 Bama miniature pigs, 1 died from transplantation failure intraoperatively. The operation time of the remaining 5 pigs was (247±27) min and the time of anhepatic phase was (46±4) min. Three animals survived for more than 2 weeks. Compared with the preanhepatic phase, the heart rate of the animals was significantly faster, MAP was considerably reduced to (46±6) mmHg, blood pH value, base excess (BE) and HCO₃^- level were all significantly decreased and serum level of K⁺ was significantly elevated during the anhepatic phase (all P < 0.05). In the neohepatic phase, MAP of Bama miniature pigs was significantly increased, heart rate was dramatically slower.Blood pH value, BE, HCO₃^- level were significantly increased and serum level of K⁺ was significantly declined (all P < 0.05). During abdominal closure, MAP, blood gas indexes and serum level of K⁺ were almost recovered to those in the preanhepatic phase. Compared with preoperative levels, the levels of alanine aminotransferase(ALT), aspartate aminotransferase(AST), lactate dehydrogenase(LDH)and alkaline phosphatase(ALP)were significantly increased after operation (all P < 0.05), the change in AST was the most obvious, and it gradually decreased at postoperative 2 d. The level of γ-gutamyl transferase(GGT) did not significantly elevated. The level of total bilirubin (TB) was evidently elevated at postoperative 5 d. Compared with the preoperative levels, the levels of total protein (TP) and albumin (ALB) were significantly decreased after operation (both P < 0.05), and began to gradually increase at postoperative 1 d. Conclusions The non-venous bypass orthotopic liver transplantation model of Bama miniature pig is convenient, with highly reproducible and survival rate, which can be utilized as a standardized liver transplantation model.

Objective To investigate the application of CT image and cone beam computed tomography (CBCT) image registration based on 3D Slicer software in image-guided radiotherapy for uterine cervical neoplasms. Methods Based on 3D Slicer software and Slicer RT toolkit, 10 positioning CT images and 50 CBCT images of 10 patients with uterine cervical neoplasms in Henan Provincial People's Hospital between January 2018 and October 2018 had rigid registration and b-spline deformation registration respectively. The dice similarity coefficient (DSC) and Hausdorff distance (HD) of the bladder, rectum, femoral head, spinal cord, and body of CT-CBCT images were compared by using paired t-test before and after the registration. Results Pre-registration, rigid registration and after b-spline deformation registration of CT images and CBCT images, the DSC in the bladder (0.459±0.177, 0.528±0.184, 0.542±0.187, respectively), the rectum (0.564±0.141, 0.632±0.091, 0.684±0.097, respectively), the femoral head (0.695±0.088, 0.833± 0.030, 0.865±0.027, respectively), the spinal cord (0.587±0.119, 0.746±0.085, 0.834±0.032, respectively) and the body surface (0.922±0.013, 0.948±0.011, 0.959±0.009, respectively) showed an increased trend; HD in the bladder (12.8±7.2, 12.2±7.1, 11.7±7.3, respectively), the rectum (5.0±1.8, 4.4±1.2, 3.4±1.2, respectively), the femoral head (3.6±1.2, 1.8±0.5, 1.5 ±0.5, respectively), the spinal cord (4.0 ±1.0, 2.7 ±1.3, 1.8 ±0.5, respectively) and the body surface (6.3±2.1, 5.2±2.0, 4.3±2.0, respectively) showed a decreased trend. The differences of pairwise comparison in the same parts were statistically significant (all P < 0.05). Conclusions Both rigid registration and b-spline deformation registration of CT-CBCT images based on 3D Slicer softwarecan improve the radiotherapy accuracy of uterine cervical neoplasms, and b-spline deformation registration has more significant advantages.

A case of late-onset methylmalonic aciduria combined with hyperhomocysteinemia (cblC type) is reported. The main manifestations were the reduction of intelligence,the instability of walking,and the inability to take care of oneself,with secondary cerebral hemorrhage. The effect of treatment was good. MMACHC gene mutation detection showed exon1 deletion, indicating that delExon1 is one of the causes of late onset methylmalonic aciduria, cblC type.

Objective To study the efficacy and safety of intravesical chemotherapy combined with intravenous chemotherapy for high grade T1 (T1G3) bladder cancer after transurethral resection of bladder tumor(TURBT).Methods From January 2012 to December 2015,111 patients with high grade T1 (T1 G3) bladder cancer were retrospectively reviewed.Thirty-six patients received TURBT and intravesical chemotherapy and intravenous chemotherapy (group A),75 patients received TURBT and intravesical chemotherapy(group B).In group A,there were 28 males and 8 females,with average age 66.2 years;in group B,there were 59 males and 16 females,with mean age 67.9 years.There was no statistical difference between the two groups in age,sex,smoking history,tumor diameter,tumor number.1-year recurrence-free survival (RFS),1-year progression-free survival (PFS),intravenous chemotherapy adverse reaction were analyzed.Results All the patients were followed-up for 12 months.9 patients relapsed in group A,1-year RFS rate 75％,and the median RFS of the 9 patients was 9 (3-11) months.36 patients relapsed in group B,1-year RFS rate 52％,and the median RFS of the 36 patients was 7 (3-11) months.There was statistically significant difference between the two group(P =0.02).One patient progressed in group A,1-year PFS rate 97.2％,and the PFS was 9 months.Six patients progressed in group B,1-year PFS rate 92％,and the median PFS was 9.5(6-12) months.There was no statistically significant difference (P =0.305) between the two group.Only 1 case (3％) appeared Ⅲ° or above intravenous chemotherapy adverse reaction.Conclusions Intravesical chemotherapy combined with intravenous chemotherapy offers a better RFS rate than the intravesical chemotherapy alone for patients with T1G3 bladder cancer after TURBT,and there are very low rates of serious side effects.Intravenous chemotherapy may be considered as a new therapy strategy for T1G3 bladder cancer after TURBT.

OBJECTIVE:To investigate the efficacy and safety of Xiaojin pill combined with docetaxel,epirubicin and cyclo-phosphamide in the treatment of breast cancer in stage Ⅲ and its effect on immune functions. METHODS:102 patients with breast cancer in stage Ⅲ were randomly divided into control group (51 cases) and observation group(51 cases). All patients were given symptomatic treatments as hydration,alkalization and antiemetic treatments in the duration of chemotherapy,and were given conven-tional treatments as 0.75 mg Dexamethasome oral tablets in 12 and 6 hours before the chemotherapy lespectively. Control group was given 75 mg/m2 Docetaxel injection,d1,intravenous infusion+60 mg/m2 Epirubicin hyclrochloride for injection,d1,intravenous in-fusion+600 mg/m2 Cyclophosphamide for injection,d1,intravenous infusion;observation group was additionally given 6 g Xiaojin pill,orally,twice a day. 21 d was regarded as a treatment course. Short-term clinical efficacy,immunoglobulin,T lymphocyte sub-sets(CD4+/CD3+,CD8+/CD3+,CD4+/CD8+)and the incidence of adverse reactions before and after chemotherapy in 2 groups after 2 courses were observed. RESULTS:There were no significant differences in the short-term effective rate and incidence of adverse reactions between 2 groups (P>0.05). Before chemotherapy,there were no significant differences in the immunoglobulin and T lymphocyte subsets between 2 groups(P>0.05). After chemotherapy,immunoglobulin levels in 2 groups were significantly lower than before,the differences were statistically significant (P<0.05);but there were no significant differences between 2 groups (P>0.05);CD4+/CD3+,CD8+/CD3+,CD4+/CD8+ in control group and CD8+/CD3+ in observation group were significantly lower than before,and CD4+/CD3+ and CD4+/CD8+ in observation group were higher than control group,the differences were statistically significant (P<0.05),but there was no significant difference in the CD8+/CD3+ between 2 groups (P>0.05). CONCLUSIONS:Based on the conventional treatment,Xiaojin pill combined with docetaxel,epirubicin and cyclophosphamide shows significant effi-cacy in the treatment of breast cancer in stageⅢ,and can improve immune functions,with good safety.

PURPOSE: Allergic rhinitis (AR) is an inflammatory disorder of the upper airway. Exosomes or extracellular vesicles are nanosized vesicles of endosomal origin released from inflammatory and epithelial cells that have been implicated in allergic diseases. In this study, we characterized the microRNA (miRNA) content of exosomes in AR. METHODS: Extracellular vesicles were isolated from nasal mucus from healthy control subjects (n=10) and patients with severe AR (n=10). Vesicle RNA was analyzed by using a TaqMan microRNA assays Human Panel-Early Access kit (Applied Biosystems, Foster City, CA, USA) containing probes for 366 human miRNAs, and selected findings were validated with quantitative RT-PCR. Target prediction and pathway analysis for the differentially expressed miRNAs were performed using DIANA-mirPath. RESULTS: Twenty-one vesicle miRNAs were up-regulated and 14 miRNAs were under-regulated significantly (P<0.05) in nasal mucus from AR patients when compared to healthy controls. Bioinformatic analysis by DIANA-mirPath demonstrated that 32 KEGG biological processes were significantly enriched (P<0.05, FDR corrected) among differentially expressed vesicle miRNA signatures. Among them, the B-cell receptor signaling pathway (P=3.709E-09), the natural killer cell-mediated cytotoxicity (P=8.466E-05), the T-cell receptor signaling pathway (P=0.00075), the RIG-I-like receptor signaling pathway (P=0.00127), the Wnt signaling pathway (P=0.00130), endocytosis (P=0.00440), and salivary secretion (P=0.04660) were the most prominent pathways enriched in quantiles with differential vesicle miRNA patterns. Furthermore, miR-30-5p, miR-199b-3p, miR-874, miR-28-3p, miR-203, and miR-875-5p, involved in B-cell receptor and salivary secretion signaling pathways, were selected for validation using independent samples from 44 AR patients and 20 healthy controls. MiR-30-5p and miR-199b-3p were significantly increased in extracellular vesicles from nasal mucus when compared to healthy controls, while miR-874 and miR-28-3p were significantly down-regulated. In addition, miRNA-203 was significantly increased in AR patients, while miRNA-875-5p was found to be significantly decreased in AR patients. CONCLUSIONS: This study demonstrated that vesicle miRNA may be a regulator for the development of AR.
B-Lymphocytes ; Biological Processes ; Endocytosis ; Epithelial Cells ; Exosomes ; Humans ; MicroRNAs* ; Mucus* ; Receptors, Antigen, T-Cell ; Rhinitis* ; RNA ; Wnt Signaling Pathway