OBJECTIVE: To study the effects of Anxin Granules on dyslipidemia in rabbits caused by high fat plus high cholesterol diet. METHODS: Thirty-two healthy New Zealand male white rabbits were randomly divided into 4 groups: normal control group, untreated group, Zhibituo Tablet-treated group and Anxin Granule-treated group. Rabbits in the normal control group were fed with regular chow, while rabbits in the other three groups were fed with high fat plus high cholesterol diet. Zhibituo Tablets and Anxin Granules were administered to the rabbits in Zhibituo Tablet-treated group and Anxin Granule-treated group at a daily oral dose respectively. At the end of the 10th week, the levels of serum total cholesterol (TC), triglycerides (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apolipoprotein A(1) (ApoA1) and apolipoprotein B (ApoB) were tested in each group, and the ultrastructures of the aorta were also observed by an electron microscope. RESULTS: Anxin Granules could reduce the levels of TC, TG, LDL-C and ApoB. The results observed by electron microscope showed that, as compared with the untreated group and the Zhibituo Tablet-treated group, the atherosclerosis of aorta in the Anxin Granule-treated group was lighter. And it was found that there were few lipid droplet vacuoles in cytoplasm of the endothelial cells, and various cell organs and elastic membrane were existed, but no lipid droplet vacuoles in cytoplasm of the medial smooth muscle cells. CONCLUSION: Anxin Granules can regulate the metabolism of blood lipid and inhibit the formation of atherosclerosis caused by hyperlipidemia in rabbits.

Objective To compare the short-term effect of laparoscopic surgery with open surgery for gastrointesti﹣nal stromal tumor. Methods 26 patients, whose tumor were localized and the pathologic diagnosis were gastrointesti﹣nal stromal tumor in stomach after surgery, were randomly divided into laparoscopic surgery group (n=12) and open surgery group (n= 14). The age and gender level, disease duration, clinical symptom, tumor location, and the lesion diameter were similar between the two groups (P> 0.05). Surgery achievement, intraoperative blood loss, operative time, postoperative hospital stay, postoperative flatus time, postoperative liquid time, and incidence rates of postoper﹣ative complications were compared between the two groups. Results All the patients get successful R0 resection. The volume of intraoperative blood loss, operative time, postoperative hospital stay, postoperative flatus time, and postop﹣erative liquid time in laparoscopic surgery group were lower than open surgery group (P< 0.05). There was no sig﹣nificant difference in the incidence rate of postoperative complications between the two groups (P> 0.05). Conclusion Compared with open surgery, the short-term effect of laparoscopic surgery were better for patients whose gastrointestinal stromal tumor in stomach were localized, and it is worth of promoting clinical application.

OBJECTIVE: To study the assessed value of 64 slice spiral CT perfusion imaging (CTPI) in laryngeal squamous cell carcinoma after chemotherapy and radiotherapy. METHOD: Forty five patients diagnosed with local advanced laryngeal squamous cell carcinoma were selected. Conventional CT and CTPI were performed before treatment and at the time of radiation dose up to 40 Gy. Blood flow, blood volume, mean transit time and surface permeability were measured at the same time. According to the decrease of tumor volume in final examination, patients were divided into sensitive group and insensitive group. The tumor perfusion indexes were compared between groups. RESULT: Blood flow, blood volume, surface permeability after 40Gy treatment were lower than before treatment in both sensitive group and the insensitive group ascended(P<0. 05). The AUC of ROC of blood flow, blood volume, mean transit time and surface permeability were 0. 804, 0. 843, 0. 852 and 0. 826. The sensitivity, specificity and accuracy of blood flow was 89. 7%, 86.8% and 90. 9%. There were 100. 0%, 91. 4% and 93. 7% in blood volume; 100. 0%, 67. 7% and 88. 3% in mean transit time; 91. 2%, 69. 4% and 90. 6% in surface permeability(P<0. 01). CONCLUSION Sixty-four slice spiral CT perfusion imaging is able to assess tumor status of laryngeal squamous cell carcinoma after chemotherapy and radiotherapy effectively.
Carcinoma, Squamous Cell ; diagnosis ; drug therapy ; radiotherapy ; Head and Neck Neoplasms ; diagnosis ; drug therapy ; radiotherapy ; Humans ; Laryngeal Neoplasms ; diagnosis ; drug therapy ; radiotherapy ; Perfusion Imaging ; Sensitivity and Specificity ; Squamous Cell Carcinoma of Head and Neck ; Tomography, Spiral Computed ; Tumor Burden

Objective: To observe the effects of fecal microbiota transplantation (FMT) on acute lung injury (ALI) induced by lipopolysaccharide (LPS) in rats and to investigate the possible mechanism in order to provide new thoughts for the treatment of acute lung injury. Methods: Forty-five adult healthy male SPF level SD rats were randomly divided into three groups of normal saline (NS), LPS and FMT with 15 in each group. ALI was induced by intraperitoneal injection of rats with 5 mg/kg of LPS. The FMT group was given 10 ml/kg of fecal microbiota solution intervention (twice a day for two consecutive days) after ALI induction. Five rats in each group were randomly selected and sacrificed at 24 h, 48 h and 72 h after intervention. Pathological changes in lung tissues were examined with hematoxylin and eosin (HE) staining and pathological scores were assessed. Right lung samples were weighed to measure wet/dry (W/D) ratios. Abdominal aorta blood samples were collected for PaO₂ analysis. The levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) and interleukin 6 (IL-6) in bronchoalveolar lavage fluid (BALF) were detected by enzyme linked immunosorbent assay (ELISA). Expression of transforming growth factor-β (TGF-β) and intracellular signal transduction protein Smads (Smad3 and Smad7) at mRNA level was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). Expression of ERK1/2 and p-ERK1/2 at protein level was detected by Western blot (WB). Rat fecal samples were collected to extract DNA and the PCR products of V3 and V4 regions were sequenced by Illumina MiSeq. Bioinformatics analysis on microbiota was conducted with Illumina MiSeq based on operational taxonomic unit (OTU) clustering. Results: Compared with the NS group, the LPS group showed significantly widened alveolar septa, massive inflammatory cell infiltration and some alveolar collapse in lung tissues. Compared with the LPS group, the FMT group showed significantly alleviated inflammatory cell infiltration, reduced swelling in pulmonary interstitial tissues, and less damage to pulmonary structure. Compared with the NS group, the W/D lung ratio, PaO₂ concentration and TNF-α, IL-1 and IL-6 levels in BALF in the LPS group significantly increased at 24 h after intervention and then gradually decreased, but still higher than those in the NS group (P<0.05). The W/D lung ratio, PaO2 concentration and TNF-α, IL-1 and IL-6 levels in BALF of the FMT group were significantly lower than those of the LPS group (P<0.05). Compared with the NS group, the expression of TGF-β1 and Smad3 at mRNA level in lung tissues increased at all time points in the LPS group, while the expression of Smad7 at mRNA level decreased (P<0.05). The expression of TGF-β1 and Smad3 at mRNA level in the FMT group were significantly lower than that in the LPS group (P<0.05), while the expression of Smad7 at mRNA level was higher (P<0.05). Compared with the NS group, p-ERK expression significantly increased in the LPS group with the highest expression at 24 h and gradually decreased at 48 h and 72 h (P<0.05). The expression of p-ERK in the FMT group was significantly lower than that in the LPS group (P<0.05). Results of the intestinal microbiota sequencing revealed 1 362 OTU in the NS group, 1 443 OTU in the LPS group and 1 510 OTU in the FMT group. There were 336 OTU shared by them accounting for 26.4%. The constitution of intestinal microbiota in the LPS group was significantly different with that of normal rats and characterized by high diversity with increased Firmicutes/Bacteroidetes ratio and decreased gene abundance of intestinal microbiota. After the intervention of fecal bacteria transplantation, the constitution of intestinal microbiota in the FMT group was similar to that of the NS group, showing decreased Firmicutes/Bacteroidetes ratio and increased gene abundance of the bacterial community (P<0.05). Conclusions FMT might inhibit immune inflammation, reduce the production and secretion of inflammatory markers in the body, alleviate alveolar epithelial damage and abnormal repair through regulating intestinal microbiota and TGF-β1/Smads/ERK pathway to improve the LPS-induced endotoxic ALI in rats.

Objective:To observe the efficacy and safety of Tofacitinib in treating elderly rheumatoid arthritis(RA), in order to provide clinical evidence.Methods:In the randomized control trial, a total of 90 elderly RA patients admitted to the Department of Rheumatology of the First Affiliated Hospital of Soochow University from January 2019 to January 2021 were selected and divided into Methotrexate group(MTX group, MTX 10mg, qw, n=45)and Tofacitinib group(TOF group, oral 5mg, bid, n=45). The efficacy and safety of the two groups were evaluated at week 12.The primary endpoint was the proportion of patients meeting the American College of Rheumatology 50%(ACR50)improvement response criteria at week 12.Secondary endpoints included ACR20/70 improvement response, proportion of patients who met treat-to-target(T2T)criteria, including Disease Activity Score in 28 joints using erythrocyte sedimentation rate(DAS28-ESR), Disease Activity Score in 28 joints using C-reactive protein level(DAS28-CRP), clinical disease activity index(CDAI), and simplified disease activity index(SDAI), and patient-reported outcomes(PROs)which included changes compared to baseline in pain visual analog scale(VAS)and Health Assessment Questionnaire Disability Index(HAQ-DI)score, at week 12.Safety outcomes including drug-related adverse events, serious adverse events, dropping out due to adverse events, and deaths were assessed throughout.Results:Five patients in each group withdrew from the trial due to adverse events, and the number of patients who finally completed the observation was 40 in each group.At week 12, the ACR50 response rate was higher in TOF group than in MTX group[35%(14/40) vs.12.5%(5/40), χ2=5.591, P=0.018)], achieving the primary endpoint.When comparing TOF vs.MTX group, the ACR20 response rate[55%(22/40) vs.25%(10/40), χ2=7.500, P=0.006]and ACR70 response rate[25%(10/40) vs.7.5%(3/40), χ2=4.501, P=0.034], and proportions of indexes of disease remission including DAS28-ESR<2.6[25%(11/40) vs.7.5%(3/40), χ2=4.501, P=0.034], or DAS28-CRP<2.6[27.5%(11/40) vs.7.5%(3/40), χ2=5.541, P=0.019], or CDAI≤2.8[30%(12/40) vs.10%(4/40), χ2=5.000, P=0.025], or SDAI≤3.3[27.5%(11/40) vs.7.5%(3/40), χ2=5.541, P=0.019], and the proportions of patients with low disease activity including DAS28-ESR≤3.2[32.5%(14/40) vs.12.5%(5/40), χ2=5.591, P=0.018], or DAS28-CRP≤3.2[32.5%(14/40) vs.12.5%(5/40), χ2=5.591, P=0.018], or CDAI≤10[37.5%(15/40) vs.17.5%(7/40), χ2=4.013, P=0.045], or SDAI≤11[37.5%(15/40) vs.15%(6/40), χ2=5.230, P=0.022], as well as changes compared to baseline data in pain VAS[(26.51±8.32)scores vs.(14.16±4.39)scores, t=8.371, P<0.001]and in HAQ-DI score(0.65±0.24 vs.0.32±0.06, t=9.387, P<0.001)were all better in the TOF group than in the MTX group at week 12.During the 12-week observation period, the number of patients with infection and hyperlipidemia was higher in TOF group than in MTX group, while the number of patients with abnormal blood cell count and liver function was lower than that in MTX group, but the differences were not statistically significant(all P<0.05). Conclusions:Tofacitinib has good efficacy and safety in the elderly RA.In patients over 70 years of age who are at high risk of infection, tofacitinib should be used with caution.

Objective:To explore the expression of long non-coding RNA (lncRNA) RP13-349O20.2 in cervical cancer tissues and its impact on the migration, invasion abilities and chemotherapy sensitivity of cervical cancer cells in vitro and the possible mechanisms.Methods:The GEPIA.CANCER website (the data was updated in June 2023) was used to analyze the relationship between the expression level of RP13-349O20.2 and the overall survival of 253 cervical cancer patients. From January 2020 to August 2022, cancer tissues and paracancerous tissues (>2 cm from the tumor edge) from 40 cervical cancer patients in the Affiliated Tengzhou Central People's Hospital of Xuzhou Medical University were retrospectively collected. Human normal cervical epithelial cells H8 and human cervical cancer cell lines HCC94, C33A, Hela, HCC1106 and SiHa were used for cell experiments in vitro. Real-time fluorescence polymerase chain reaction (qRT-PCR) was used to detect the relative expression of RP13-349O20.2 in cervical cancer tissues, paracancerous tissues and each cell line. The C33A cells with the highest relative expression level of RP13-349O20.2 were transfected with small interfering RNA (siRNA) of RP13-349O20.2 and siRNA of its negative control sequence, and they were si-RP13-349O20.2 group and si-Con group, respectively. The scratch healing assay was used to detect the migration ability of C33A cells in the two groups, the Transwell assay was used to detect the invasion ability of C33A cells, and the CCK-8 method was used to detect the sensitivity of C33A cells to 5-fluorouracil. The absorbance value indicated the cell proliferation ability, the lower the absorbance value, the weaker the proliferation ability, the more sensitive to the drug. Dual-luciferase reporter gene assay was used to verify the targeting relationship between RP13-349O20.2 and miRNA-493-5p (miR-493-5p), miR-493-5p and Nectin-4. qRT-PCR was used to detect the relative expression of miR-493-5p and Nectin-4 mRNA in two groups of C33A cells, and Western blotting was used to detect the expressions of Nectin-4 protein and PI3K-AKT signaling pathway proteins in two groups of cells.Results:Analysis based on data from GEPIA.CANCER website shows that patients with low expression of RP13-349O20.2 had better overall survival than patients with high expression ( P < 0.01). The relative expression levels of RP13-349O20.2 in cervical cancer tissues and paracancerous tissues of 40 patients were 4.04±0.32 and 1.18±0.14, and the difference was statistically significant ( t = 8.29, P < 0.01). Compared with H8 cells, the expressions of RP13-349O20.2 in human cervical cancer cell lines HCC94, C33A, Hela, HCC1106 and SiHa were higher (all P < 0.01). The relative expression levels of RP13-349O20.2 in C33A cells in the si-Con group and si-RP13-349O20.2 group were 7.30±0.30 and 1.01±0.27, and the difference was statistically significant ( t = 15.62, P < 0.01). The scratch healing rates of C33A cells in the si-Con group and si-RP13-349O20.2 group were (32±9)% and (75±6)% ( t = 3.97, P < 0.01), and the numbers of invasive cells were (106±12) cells and (36±8) cells ( t = 4.79, P < 0.01). After the action of 5, 10, 20, 40 and 80 μmol/L 5-fluorouracil for 24 h, the absorbance value of C33A cells in the si-RP13-349O20.2 group was lower than that in the si-Con group. Dual-luciferase reporter gene assay confirmed that there was a targeting relationship between P13-349O20.2 and miR-493-5p ( P < 0.01), and there was a targeting relationship between miR-493-5p and Nectin-4 ( P < 0.01) . The relative expression levels of miR-493-5p in C33A cells in the si-Con group and si-RP13-349O20.2 group was 1.02±0.13 and 5.48±0.85 ( t = 5.21, P < 0.01). The relative expression levels of Nectin-4 mRNA were 5.65±0.33 and 0.99±0.34 ( t = 9.87, P < 0.01). The expression of Nectin-4 protein in C33A cells in the si-RP13-349O20.2 group was lower than that in the si-Con group ( t = 9.21, P = 0.001), and the expressions of PI3K-AKT signaling pathway proteins p-STAT3, p-PI3K, p-AKT and p-mTOR were lower than those in the si-Con group (all P < 0.01). Conclusions:The level of RP13-349O20.2 in cervical cancer tissues is high, and its high expression may indicate the poor prognosis of patients. Interfering with the expression of RP13-349O20.2 in vitro can inhibit the migration and invasion abilities of cervical cancer cells and promote the sensitivity of cervical cancer cells to 5-fluorouracil. The mechanism may be related to the miR-493-5p/Nectin-4 signaling pathway and the PI3K-AKT signaling pathway.

Objective To investigate effects of polyphylin Ⅰ on the proliferation and apoptosis of human melanoma cell line A375,and to explore their mechanisms.Methods Normal human melanocytes isolated from healthy human foreskin were divided into 6 groups to be treated with 0,1.5,3.0,6.0,9.0,12.0 mg/L polyphyllin Ⅰ respectively.A375 melanoma cells were divided into 4 groups,i.e.,control group,1.5-,3.0-,6.0-mg/L polyphyllin Ⅰ groups,to be treated with 0,1.5,3.0,6.0 mg/L polyphyllin Ⅰ,respectively.Cell counting kit-8 (CCK8) assay was performed to evaluate the effect of polyphyllin Ⅰ on the proliferation of normal human melanocytes and A375 cells.Hoechst 33258 fluorescent staining was conducted to observe the morphology of apoptotic cells,flow cytometry to estimate cell cycle phase distribution and apoptosis rate,dichloro-dihydro-fluorescein diacetate (DCFH-DA) fluorescent probe assay to detect the level of reactive oxygen species (ROS),rhodamine-123 staining to evaluate changes of mitochondrial membrane potential,spectrophotography to detect the level of ATP in A375 cells,as well as levels of lactic acid and glucose in the culture supernatant of A375 cells,and Western blot analysis to determine the protein expression of Bcl-2,Bcl-2-related X protein (Bax),cleaved-caspase-3,cyclin D1 and pyruvate kinase isozyme type M2 (PKM2).Statistical analysis was carried out by using one-way analysis of variance (ANOVA) for comparisons among groups and Student-Newman-Keuls-q (SNK-q) test for multiple comparisons.Results CCK8 assay showed that the treatment with polyphyllin Ⅰ at concentrations of 1.5,3.0,6.0 mg/L for 48 hours had no effects on the proliferation of normal human melanocytes,but significantly inhibited the proliferation of A375 cells.The survival rate of A375 cells was significantly lower in the 1.5-,3.0-,6.0-mg/L polyphyllin Ⅰ groups than in the control group (P ＜ 0.01).After the treatment with polyphyllin Ⅰ,distinct apoptotic morphology of A375 cells was observed under fluorescence microscope.Additionally,along with the increase of polyphyllin Ⅰ concentrations (0,1.5,3.0,6.0 mg/L),there were gradual increasing trends in the apoptosis rate of A375 cells (4.25％ ± 1.27％,10.03％ ± 1.49％,36.62％ ± 1.97％,44.11％ ± 2.47％ respectively,F =665.7,P ＜ 0.01),the percentage of A375 cells at G0/G1 phase (54.13％ ± 2.57％,67.35％ ± 3.79％,74.39％ ± 3.29％,82.29％ ± 3.99％ respectively,F =71.81,P ＜ 0.01),the level of ROS in A375 cells (P ＜ 0.01),the level of glucose in the culture supernatant (P ＜ 0.01),and the protein expression of Bax and cleaved-caspase-3 (both P ＜ 0.01),while gradual decreasing trends were found in the levels of mitochondrial membrane potential and ATP in A375 cells (both P ＜ 0.01),the level of lactic acid in the culture supernatant (P ＜ 0.01),and the protein expression of Cyclin D1,Bcl-2 and PKM2 (all P ＜ 0.01).Conclusion Polyphyllin Ⅰ can effectively induce A375 cell apoptosis by promoting the production of ROS in A375 cells and decreasing the mitochondrial membrane potential,and arrest A375 cells at G0/G1 phase by inhibiting the expression of PKM2 and Cyclin D1.