Background Imidacloprid is a neonicotinoid insecticide that is widely used in agricultural production, with a high detection rate in human biological samples. Previous studies have shown a high correlation between imidacloprid exposure and liver injury, but the specific mechanism is still unknown. Objective To observe potential toxic effects of HepG2 cells and its perturbation of non-targeted metabolic profile after imidacloprid exposure, and to explore possible molecular mechanisms of hepatotoxicity of imidacloprid by analyzing invovlved biological processes and signaling pathways. Methods HepG2 cell suspension was prepared and seeded in a 96-well plate, which was divided into blank control group, dimethyl sulfoxide (DMSO) solvent control group and imidacloprid exposure groups with multiple concentrations. Each group was set with 5 parallel samples. The viability of HepG2 cells viability were determined after 8 h of exposure to different concentrationsof imidacloprid (1, 2.5, 5, 7.5, 10 mmol·L⁻¹), and the dose-effect relationship was analyzed. A proper concentration (3 mmol·L⁻¹ with 80% viability) was chosen for imidacloprid exposure, non-targeted metabolomic analysis was applied to the cultivated HepG2 cells using UHPLC-Q-TOF/MS technology, the differential metabolites between groups were screened, and the bioprocess and related signaling pathways of their enrichment were annotated using the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. Results Compared to the other two groups, the survival rates of HepG2 cells in the imidacloprid exposure groups decreased. A survival rate of about 86% of HepG2 cells was found in HepG2 cells exposed to 2.5 mmol·L⁻¹ imidacloprid exposure. The non-targeted metabolomics studies showed that 61 metabolites were significantly affected in HepG2 cells after 3 mmol·L⁻¹ imidacloprid exposure, including creatine (variable importance in projection VIP=1.11, P<0.001), arginine (VIP=1.47, P=0.048), taurine (VIP=4.28, P=0.001), and α-D-glucose (VIP=1.90, P=0.006). The differential metabolites enriched in bioprocess and related signaling pathways were mainly directed to mTOR signaling pathways (P<0.001), arginine and proline metabolism (P=0.002), and galactose metabolism (P=0.015). Conclusion Imidacloprid exposure can significantly inhibit the survival rate of HepG2 cells, and interfere with the mTOR signaling pathway, arginine and proline metabolism, galactose metabolism, and so on.

Objective To explore the improvements of high-fat intake on lung injury induced by Paragonimus proliferus infection in rats, and to preliminarily explore the mechanisms underlying the role of cytochrome P450 4A1 (CYP 4A1) in the improve ments. Methods SD rats were randomly assigned into three groups, including the normal control group (n = 10), the infection and normal diet group (n = 12) and the infection and high-fat diet group (n = 12). Rats in the normal control group were fed with normal diet and without any other treatments, and animals in the infection and normal diet group were subcutaneously injected with 8 excysted metacercariae of P. proliferus via the abdominal wall, followed by feeding with normal diet, while rats in the infection and high-fat diet group were subcutaneously injected with 8 excysted metacercariae of P. proliferus via the abdominal wall, followed by feeding with high-fat diet. All rats were sacrificed 28 weeks post-infection, and serum samples and lung specimens were collected. Following hematoxylin-eosin (HE) staining of rat lung specimens, the rat lung injury was observed under an optical microscope, and alveolitis was evaluated using semi-quantitative scoring. Serum interleukin-1β (IL-1β) and tumor necrosis factor alpha (TNF-α) levels were measured using enzyme-linked immunosorbent assay (ELISA), and the cytochrome P450 4A1 (CYP 4A1) expression was quantified in rat lung specimens at transcriptional and translational levels using quantitative real-time PCR (qPCR) and Western blotting assays. Results Alveolar wall thickening, edema and inflammatory cell infiltration were alleviated 28 weeks post-infection with P. proliferus in rats in the infection and high-fat diet group relative to the infection and normal diet group, and no alveolar consolidation was seen in the infection and high-fat diet group. The semi-quantitative score of alveolitis was significantly higher in the infection and normal diet group [(2.200 ± 0.289) points] than in the normal control group [(0.300 ± 0.083) points] and the infection and high-fat diet group [(1.300 ± 0.475) points] (both P values < 0.05), and higher serum IL-1β [(151.586 ± 20.492)] pg/mL and TNF-α levels [(180.207 ± 23.379) pg/mL] were detected in the infection and normal diet group than in the normal control group [IL-1β: (103.226 ± 3.366) pg/mL; TNF-α: (144.807 ± 1.348) pg/mL] and the infection and high-fat diet group [IL-1β: (110.131 ± 12.946) pg/mL; TNF-α: (131.764 ± 27.831) pg/mL] (all P values < 0.05). In addition, lower CYP 4A1 mRNA (3.00 ± 0.81) and protein expression (0.40 ± 0.02) was quantified in lung specimens in the infection and normal diet group than in the normal control group [(5.03 ± 2.05) and (0.84 ± 0.14)] and the infection and high-fat diet group [(11.19 ± 3.51) and (0.68 ± 0.18)] (all P values < 0.05). Conclusion High-fat intake may alleviate lung injuries caused by P. proliferus infection in rats through up-regulating CYP 4A1 expression in lung tissues at both translational and transcriptional levels.

Psoriasis is considered as an inflammatory disease driven by T cells, and its pathogenesis is closely related to the imbalance of intestinal bacteria flora. It has been reported that Bacteroides fragilis could play an anti-inflammatory role by regulating the expression of cytokines in T cells. To date, there is no report using B. fragilis to treat psoriasis. In this study, we explored the therapeutic effect of B. fragilis BF839 on psoriasis. We selected 27 psoriasis patients who were treated in the Second Affiliated Hospital of Guangzhou Medical University from April to October 2019. The patients were given B. fragilis BF839 orally for 12 weeks while maintaining the original treatment. The psoriasis area and severity index (PASI) score was evaluated before and after the treatment. The rate of drug withdrawal and reduction after 12 weeks of treatment were calculated. Our results showed that the rate of 12-week trial completion was 96.3% (26/27). We used PASIN to define the proportion of people whose PASI score decreased more than or equal to N% after treatment. At 12 weeks, PASI30, PASI50, and PASI75 were 65.4%, 42.3%, and 19.2%, respectively. The PASI score was 9.1±5.9 and 5.8±4.9 before and after 12 weeks of treatment respectively, and the difference was statistically significant (P<0.01). The effective rate of the visual analog scale (VAS) score was 42.3% at 12 weeks, and the VAS score was 2.9±2.2 and 2.3±2.1 before and after 12 weeks of treatment, respectively, which had no statistically significant difference (P>0.05). The adverse reaction rate of patients was 3.8% (1/26) within 12 weeks of treatment, including 1 case of constipation, and the rate of drug withdrawal and reduction was 60.0%. The above results suggest that B. fragilis BF839 may be functional on the treatment of psoriasis by reducing the PASI score and the drug usage rate with few side effect, which deserves further study.
Anti-Inflammatory Agents ; Bacteroides fragilis ; Cytokines ; Humans ; Psoriasis/drug therapy* ; Severity of Illness Index ; Treatment Outcome

Objective: To explore the clinical efficacy and prognostic factors of first-generation and second-generation tyrosine kinase inhibitors (TKI) based regimen in the treatment of patients with BCR-ABL positive acute lymphoblastic leukemia (ALL) . Methods: Retrospectively analyze the clinical characteristics and prognostic factors of 89 patients with BCR-ABL positive ALL from April 2012 to June 2018 in our hospital, the clinical efficacy of first-generation and second-generation TKI was compared. Results: 60 patients were classified into the first-generation TKI (imatinib) group, and 29 patients were in the second-generation TKI (dasatinib) group. There were no significant differences in gender, age, WBC, hemoglobin concentration, PLT, chromosomal karyotype, the types of fusion genes, allogeneic hematopoietic stem cell transplantation (allo-HSCT) and TKI initiation time between the two groups. The first-generation and second-generation TKI groups, for which the complete remission (CR) rate at the fourth week of induction therapy was 83.3% and 89.7% (P=0.637) , respectively, and the complete molecular remission (CMR) was 48.3%and 58.6% (P=0.363) , respectively, the difference was not statistically significant. The 2-year overall survival (OS) rate of first-generation and second-generation TKI group was 34.9% and 64.0% (χ²=4.743, P=0.029) , the 2-year relapse free survival (RFS) rate was 17.2% and 55.0% (χ²=8.801, P=0.003) , respectively. Multivariate analysis showed that complete molecular remission (HR=0.281, 95%CI 0.151-0.523, P<0.001) was independent favorable prognostic factor for overall survival (OS) , complete molecular remission (HR=0.209, 95%CI 0.112-0.390, P<0.001) and second-generation TKI (HR=0.318, 95%CI 0.158-0.641, P=0.001) were independent favorable prognostic factors for RFS. Conclusion For TKI-based regimen of BCR-ABL positive ALL, second-generation TKI is superior to first-generation TKI in OS and RFS time.

Background: The mitogen-activated extracellular signal-regulated kinase 1/2 (MEK1/2) inhibitor trametinib has shown promising therapeutic effects on melanoma, but its efficacy on colorectal cancer (CRC) is limited. Synthetic lethality arises with a combination of two or more separate gene mutations that causes cell death, whereas individual mutations keep cells alive. This study aimed to identify the genes responsible for resistance to trametinib in CRC cells, using a synthetic lethal short hairpin RNA (shRNA) screening approach. Methods: We infected HT29 cells with a pooled lentiviral shRNA library and applied next-generation sequencing to identify shRNAs with reduced abundance after 8-day treatment of 20 nmol/L trametinib. HCT116 and HT29 cells were used in validation studies. Stable ring finger protein 183 (RNF183)-overexpressing cell lines were generated by pcDNA4-myc/his-RNF183 transfection. Stable RNF183-knockdown cell lines were generated by infection of lentivi-ruses that express RNF183 shRNA, and small interference RNA (siRNA) was used to knock down RNF183 transiently. Quantitative real-time PCR was used to determine the mRNA expression. Western blotting, immunohistochemical analysis, and enzyme-linked immunosorbent assay (ELISA) were used to evaluate the protein abundance. MTT assay, colony formation assay, and subcutaneous xenograft tumor growth model were used to evaluate cell proliferation. Results: In the primary screening, we found that the abundance of RNF183 shRNA was markedly reduced after treatment with trametinib. Trametinib induced the expression of RNF183, which conferred resistance to drug-induced cell growth repression and apoptotic and non-apoptotic cell deaths. Moreover, interleukin-8 (IL-8) was a downstream gene of RNF183 and was required for the function of RNF183 in facilitating cell growth. Additionally, elevated RNF183 expression partly reduced the inhibitory effect of trametinib on IL-8 expression. Finally, xenograft tumor model showed the synergism of RNF183 knockdown and trametinib in repressing the growth of CRC cells in vivo. Conclusion: The RNF183-IL-8 axis is responsible for the resistance of CRC cells to the MEK1/2 inhibitor trametinib and may serve as a candidate target for combined therapy for CRC.

Objective To measure the expressions of programmed death?1(PD?1)and programmed death ligand?1(PD?L1)on peripheral blood T lymphocytes of patients with condyloma acuminatum(CA), and to investigate their role in cellular immunity in these patients. Methods Peripheral blood samples were obtained from 30 patients with CA(CA group)and 20 healthy human controls (control group). Flow cytometry was conducted to detect the expressions of PD?1 and PD?L1 on the surfaces of peripheral blood CD4+and CD8+T lymphocytes, and to determine the counts of CD4+and CD8+T lymphocytes. Enzyme?linked immunosorbent assay(ELISA)was performed to measure the levels of serum interleukin?2(IL?2)and interferon?γ(IFN?γ). Statistical analyses were carried out to compare the above parameters between the two groups, and to assess the relationship of PD?1 and PD?L1 expressions with the counts of CD4+and CD8+T lymphocytes as well as with the serum levels of IL?2 and IFN?γ. Results There was a significant increase in the expression rates of PD?1 and PD?L1 on CD4+T lymphocytes(PD?1:9.48%± 3.31%vs. 7.12%± 2.16%, t=2.81, P<0.01;PD?L1:4.40%± 1.46%vs. 3.26%± 1.13%, t=3.16, P<0.01)and CD8+T lymphocytes(PD?1:12.52%± 3.17%vs. 9.95%± 2.17%, t=3.16, P<0.01;PD?L1:7.07%± 2.23%vs. 5.39%± 1.69%, t=2.88, P<0.01)in the CA group compared with the control group. Moreover, the CA group showed significantly lower counts of CD4+T lymphocytes(727.43 ± 138.59/μl vs. 804.25 ± 92.83/μl, t=2.17, P<0.05)and CD4/CD8 ratio(1.23±0.35 vs. 1.46 ± 0.34, t = 2.24, P < 0.05) than the control group, while no significant difference was observed in CD8 + T lymphocyte counts between the CA group and control group(613.60 ± 121.60/μl vs. 572.45 ± 103.08/μl, t=1.24, P>0.05). The levels of serum IL?2 and IFN?γwere both lower in the CA group than in the control group(t=2.12, 2.16, respectively, both P < 0.05). In the CA group, PD?1 and PD?L1 expression levels on peripheral blood CD4 + T lymphocytes were both negatively correlated with CD4+T lymphocyte counts, the CD4/CD8 ratio, as well as IL?2 and IFN?γserum levels(all P<0.05), and those on peripheral blood CD8+T lymphocytes were also negatively correlated with the CD4/CD8 ratio(all P<0.05), but uncorrelated with CD8+T lymphocyte counts(both P>0.05). Conclusion PD?1 was highly expressed on peripheral blood T lymphocytes from patients with CA, which may inhibit T lymphocyte?mediated immune response, decrease CD4+T lymphocyte counts, the CD4/CD8 ratio as well as IL?2 and IFN?γserum levels by interacting with its ligand PD?L1 and forming the PD?1/PD?L1 signaling pathway.

Objective To explore the long-term efficacy of botulinum toxin type A injections on facial beauty and its long-term safety.Methods A total of 33 beauty-seekers with botulinum toxin type A treatment for more than five years were reviewed as observation group,using digital muscle palpation meter (Myoton PRO) for the determination of the orbicularis oculi muscle and masseter muscle tension (F),the muscle characteristic parameters,such as muscle hardness (S) using a homemade facial questionnaire test for satisfactory rate of beauty from both beauty-seekers and physicians.At sametime,33 normal adults that never accepted botulinum toxin injection with matched age and gender were collected as control group.The same-sex indicators were dtermined and compared with using statistic analysis t test.Results The pairwised parameters of the same sex and site were comparied between the two groups;average F and S values in the observation group were lower than those of the control group,but no statistically significant difference were observed between the two groups (P＞0.05);in the observation group,average appearance age was 7.3 years younger than the control group,and the facial shape improved significantly.Conclusions Long-term and repeated application of botulinum toxin A is able to remove crow's feet and decrease the masseter and so the injection is safe with high satisfaction to beauty-seekers.

Objective To investigate the expression level of transforming growth factorβ1 (TGF-β1) and interleukin 17 (IL-17) in patients with allergic conjunctivitis. Methods TGF-β1 and IL-17 expression levels in allergic conjunctivitis patients hospitalized in the author's hospital and a healthy population were detected to explore the expression level in all types of patients with allergic conjunctivitis. Results TGF-β1 and IL-17 levels were significantly higher in patients with allergic conjunctivitis than in healthy population (P<0.01). Among the four clinical subtypes of allergic conjunctivi-tis, vernal keratoconjunctivitis had the highest expression level of TGF-β1 and IL-17 in spring, followed by perennial allergic conjunctivitis and seasonal allergic conjunctivitis, and atopic keratoconjunctivitis had the lowest expression level. However, the differences among the four subtypes were not significant (P>0.05). Conclusion Patients with allergic conjunctivitis have higher TGF-β1 and IL-17 expression levels than normal healthy people. TGF-β1 and IL-17 ex-pression level vary in different subtypes of allergic conjunctivitis, vernal keratoconjunctivitis has higher expression level than other types in spring.

Objective To investigate the effects of IL-13 on fibrosis in hepatic stellate cells and its molecular mechanism .Methods The effects of IL-13 on the proliferation of hepatic stellate cell line LX-2 were measured by MTT assay .The transcription level of collagen typeⅠ( COLⅠ) in LX-2 cells was detec-ted by RT-PCR.The secretion of COLⅠin LX-2 cells was measured by ELISA assay and hydroxyproline as-say.Western blot assay was used to analyze the effects of IL-13 on the phosphorylation of signal transducer and activator of transcription(STAT6).Results Compared with control group, IL-13 (10 ng/ml, 20 ng/ml and 50 ng/ml ) significantly stimulated the proliferation of LX-2 cells in a dose-dependent manner ( P<0.05).The expression of collagen typeⅠin LX-2 cells at mRNA and protein levels were significantly up-regulated by IL-13 at a concentration of 50 ng/ml (P<0.05), but not affected by IL-13 at low concentra-tions (5 ng/ml, 10 ng/ml, and 20 ng/ml) (P>0.05).The expression of phosphorylated STAT6 protein in LX-2 cells was significantly enhanced upon the stimulation with 50 ng/ml of IL-13 ( P<0.05 ) for60 min or 120 min.C onclusion IL-13 promoted the proliferation of human hepatic stellate cells and up-regulated the expression of COLⅠat mRNA and protein levels .IL-13 might promote the fibrosis in human hepatic stellate cells through activating STAT 6 phosphorylation .

Objective To evaluate the safety,tumor radical and early postoperative efficacy through comparison of laparoscopic gastric D2 radical surgery with traditional open gastric D2 radical surgery.Methods 254 patients with upper stomach cancer underwent surgery were selected,132 cases using conventional open gastrectomy (the traditional laparotomy group),122 patients underwent laparoscopic radical gastrectomy (laparoscopic surgery group).Laparoscopic surgery group with traditional open surgery group had no statistically significant differences in gender,age,tumor location,histological type and TNM staging.Results Open surgery group and laparoscopic surgery group had statistically significant differences in operative time [(235.78±31.56) min,(256.43±54.08) min,P ＜ 0.001],blood loss [(326.69±89.73) ml,(158.31±62.98) ml,P ＜ 0.001],incision length [(16.53±2.34) cm,(5.51±1.15) cm,P ＜ 0.001],gastrointestinal recovery time [(4.22±0.91) d,(3.31±0.83) d,P ＜ 0.001],first time eating liquid [(5.78±0.95) d,(5.56±0.78) d,P ＜ 0.001] and postoperative hospital stay [(12.62±2.89) d,(11.18±1.78) d,P ＜ 0.001].The total number of lymph node dissection and complications was not statistically significant.Conclusions Laparoscopic gastric D2 radical surgery is a safe,minimally invasive surgical method.Laparoscopic gastric D2 radical surgery has the same lymph node dissection and good early outcome compared with the traditional gastric D2 radical surgery,but postoperative recovery fast and less invasive.