[ ABSTRACT] AIM:To observe the inhibitory effect of madecassoside on the LPS-stimulated microglia and to inves-tigate its possible mechanism.METHODS:Microglia cells of neonatal Sprague-Dawley ( SD) rats were cultured, isolated and purified.Microglia cells were activated with lipopolysaccharide ( LPS) .The inhibitory effect of madecassoside on micro-glia was measured by MTT assay.Tumor necrosis factor alpha (TNF-α), interleukin 1β(IL-1β) were detected by ELISA. Cell cycle and apoptotic rate were evaluated by flow cytometry.The expression of TLR4 was detected by Western blotting. The expression of NF-κB was detected by RT-PCR.RESULTS: LPS induced the proliferation of microglia and release in-flammatory cytokines significantly.Compared with LPS group, madecassoside inhibited the proliferation of microglia induced by LPS in a dose dependent manner.The IC50 value of madecassoside was 10.97 nmol/L to microglia after incubation for 48 h.Madecassoside also decreased the levels of TNF-αand IL-6, increased the ratios of microglia at the G2 phase and the ap-optotic rate, decreased the expression of TLR4 and NF-κB significantly (P<0.05).CONCLUSION:Madecassoside has in-hibitory effects on the proliferation of LPS-stimulated microglia, by which the mechanism may be related to inhibition of the expression of TLR4 and NF-κB, change of cell cycle distribution and induction of microglia apoptosis.

Objective:To explore the categories and influencing factors of family caregivers′benefit finding in patients with esophageal cancer based on latent profile analysis.Methods:From May 2020 to February 2021, 255 primary family caregivers of patients with esophageal cancer in the First People′s Hospital of Changzhou and Affiliated Hospital of Jiangnan University were selected for the study using a cross-sectional survey method. Surveys were conducted by the General Information Questionnaire, the Benefit finding Scale, the Hospital Anxiety and Depression Scale, and the Event Related Rumination Inventory.Results:The benefit finding among family caregivers of patients with esophageal cancer was divided into 2 latent profile classifications, low benefit rumination group 33.3% (85/255) and high growth adaptation group 66.7% (170/255), and the results of binary Logistic regression analysis showed that caregivers with high school education or above ( OR=0.053, P<0.05) and higher deliberate rumination scores ( OR=0.778, P<0.01) had a higher degree of benefit finding; caregivers with higher intrusive rumination scores ( OR=1.163, P<0.05) and higher anxiety ( OR=1.323, P<0.01) and depression scores ( OR=1.128, P<0.05) had a lower benefit finding. Conclusions:There is heterogeneity in the caregiver′s benefit finding of patients with esophageal cancer. Health professionals and nursing staff should develop targeted psychological support and interventions to improve the caregiver ′s benefit finding according to the different sub-types of caregivers.

Objective To investigate the impact of micro ribonucleic acid -214 (miR-214) on the proliferation of hepatocellular carcinoma (HCC) and its mechanism. Methods MHCC97L cells were respectively transfected using miR-214 mimics and negative control mimics to establish M214 group, negative control (NC214) group. And untransfected control (Ctrl) group was established. The contents of miR-214 of MHCC97L cells in three groups were detected by fluorescence quantitative polymerase chain reaction (PCR). Cell counting kit (CCK)-8 assay was used to define the cell proliferation inhibition rate. Plate cloning formation assay was used to define the cell clonality. Cell apoptosis rate was detected by flow cytometery. The expressions of protein c-myc, Bax and B-cell lymphoma (Bcl)-2 were detected by Western blot assay. The comparison of three groups was conducted using one way analysis of variance and pairwise comparison using LSD-t test. Results The miR-214 content of MHCC97L cells in M214 group was (2 536±7) times of that in Ctrl group, where significant difference was observed (LSD-t=58.75, P<0.05). The cell proliferation inhibition rates were (15.33±0.62) %, (24.07±0.75) %, (41.02±0.91) % at the 24, 48, 72 h in M214 group, and significant difference was observed compared with that in Ctrl group (LSD-t =14.64, 19.87, 31.86; P<0.05). The clone formation quantity was (5.3±0.7) /well in M214 group, which was significantly lower compared with that in Ctrl group [(37.1±1.0)/well] (LSD-t =-12.51, P<0.05). The cell apoptosis rate was (42.1±3.2)% in M214 group, which was significantly higher compared with that in Ctrl group [(7.0±0.7) % ] (LSD-t=35.66, P<0.05). In M214 group, the expressions of protein c-myc, Bcl-2 decreased and Bax increased. Conclusion The miR-214 can inhibit the proliferation of human HCC cells through down-regulating the protein c-myc expression and Bcl-2/Bax ratio.

ObjectiveTo investigate the role of protein kinase C-epsilon (PKCε) gene silenced by small interfering ribonucleic acid (siRNA) in inhibiting the development of cholangiocellular carcinoma and its mechanism.MethodsHuman cholangiocellular carcinoma QBC939 cells were respectively transfected using PKCε-siRNA and negative control (NC)-siRNA to establish PKC group and NC group. And untransfected control (CTRL) group was established. Cell counting kit (CCK)-8 assay was used to define the cell proliferation inhibition rate. Cell apoptosis rate was detected by flow cytometery. The expressions of protein PKCε and survivin were detected by Western blot. The comparison of three groups was conducted using one way analysis of variance and pairwise comparison using LSD-t test.ResultsThe cell proliferation inhibition rates at 24, 48, 72 h [(7.52±0.33)%, (15.28±0.20)%, (37.12±0.45)% ] increased gradually in PKC group, where significant difference was observed compared with those in CTRL group (LSD-t=15.37, 27.12, 35.05;P<0.05). The cell apoptosis rate was (56.9±6.1)% in PKC group, which was significantly higher compared with that in CTRL group [(12.5±1.3) % ] (LSD-t=28.55,P<0.05). The expressions of protein PKCε and survivin decreased in PKC group compared with those in NC group and CTRL group.ConclusionPKCε gene silenced by siRNA may inhibit the development of cholangiocellular carcinoma through down-regulating the expression of protein survivin and promoting cell apoptosis.

Objective To investigate the impact of purinergic receptors P2Y12 gene inhibited by small interference RNA (siRNA) on the angiogenesis of hepatocellular carcinoma (HCC). Methods Human HCC cell line MHCC97H was infected using P2Y12-siRNA lentiviral expression vector and empty negative control vector to establish siRNA group and control group. The expression of P2Y12 protein in two groups was detected by Western blot assay. angiogenesis assay was used to deifne the angiogenesis potential of HCC cell. The microvessel density (MVD) of nude mice subcutaneous tumors was observed. The experimental data between two groups were compared using t test. Results The average relative expression of P2Y12 protein were 0.07±0.01 and 0.26±0.02 respectively in siRNA group and control group. The expression of P2Y12 protein in siRNA group decreased significantly compared with that in control group (t=-35.64, P<0.05). No continuous closed small tubular structure was observed in the human umbilical vein endothelial cell (HUVEC) stimulated by the supernatant of siRNA group, while large number of closed tubules was observed in the HUVEC stimulated by the supernatant of control group. A few microvessels of nude mice subcutaneous tumors were observed in siRNA group. The MVD was 5±1, which was signiifcantly lower than that in control group (23±6) (t=-17.01, P<0.05). Conclusion Purinergic receptors P2Y12 gene inhibited by siRNA can suppress the angiogenesis of HCC.

OBJECTIVE：To prov ide referen ce for clinical safe and rational use of belimumab by mining the risk signals of adverse drug event （ADE）. METHODS ：ADE reports related to belimumab were collected from FDA adverse event reporting system（FAERS）from the first quarter of 2015 to the first quarter of 2021. The reporting odds ratio （ROR）method and the Medicines and Healthcare Products Regulatory Agency （MHRA）method were adopted to mine the ADE risk signals related to belimumab，setting the threshold as the number of reports ＞3 and the lower limit of 95％ CI ＞1（ROR method ）and the proportional reporting ratio （PRR）＞2，and χ2 ＞4（MHRA method ）. ADEs were counted and classified by using the preferred system organ class （SOC）and preferred term （PT）of Medical Dictionary for Regulatory Activities （MedDRA）. RESULTS & CONCLUSIONS：A total of 3 529 ADE reports with belimumab as the primary suspicious drug were screened ，in which female patients（90.31％）were much more than male patients （6.15％）；age distribution was concentrated in 18-59 years old （41.80％）. There were 1 234 cases（34.97％）of severe ADE reports ，mainly involving hospital or prolonged hospital stay. Most of the reporters were consumers or other non-medical professionals （81.84％）. North America reported the most （70.39％）and the main reporting country was the United States （2 029 reports）. A total of 180 PTs were mined from 3 529 reports，in addition to PTs associated with primary disease （systemic lupus erythematosus ，pain，arthralgia，pyrexia，weight decreased ，swelling，oropharyngeal pain ， etc.），PTs related to medication error （product dose omission ，inappropriate schedule of product administration ，underdose， product availability issue ，etc.）and PTs related to infection （influenza，urinary tract infection ，infection，sinusitis，etc.）were reported in a large number of cases. Twenty-six SOCs were involved ，the top 10 SOC in ADE reports were all kinds of injuries ， poisoning and surgical complications （2 225 reports），infections and infectious diseases （1 247 reports），general disorders and administration site conditions （1 196 reports），musculoskeletal and connective tissue disorders （1 195 reports），surgical and medical procedures（515 reports），etc. PTs in SOC in the first place （all kinds of injuries ，poisoning and surgical complications ）of ADE reports were all related to medication error ；herpes zoster ，kidney infection and cellulitis in SOC in the second place （infections and infectious diseases ）of ADE reports were not included in the drug instruction of belimumab ；most PTs in SOCs such as various nervous system diseases ，immune system diseases ，mental diseases ，benign，malignant and unknown tumors （including cystic and polypoid）which were taken attention in clinic were not included in the drug instruction of belimumab. It is suggested to avoid medication errors as far as possible in clinical use of belimumab ，and to guard against adverse reactions such as herpes zoster ， kidney infection ，cellulitis and various nervous system diseases ，immune system diseases and mental diseases. In addition ，the patients with malignant tumor or related history should use belizumab carefully.