Epileptic Seizure after Use of Moxifloxacin in Man with Legionella longbeachae Pneumonia.

Legionellosis caused by Legionella longbeachae is diagnosed mainly by PCR. We report a case of L. longbeachae infection in mainland China, which was diagnosed by metagenomic next-generation sequencing, in a man who developed an epileptic seizure after using moxifloxacin. Metagenomic next-generation sequencing may be a useful tool to detect Legionella spp.

Six new furostanol glycosides from Smilax glauco-china and their cytotoxic activity.

Six new steroidal saponins, namely glauco-chinaosides A-F, and one known compound were isolated from the tubers of Smilax glauco-china. Their structures were elucidated by a combination of spectroscopic analysis and hydrolysis followed by spectral and chromatographic analysis. Compounds 1-7 were tested in vitro for their cytotoxic activities against four human tumor cell lines (SH-SY5Y, SGC-7901, HCT-116, and Lovo). Compounds 1, 2, and 5 exhibited cytotoxic activity against SGC-7901, with IC50 values of 2.7, 11.5, and 6.8 µM, respectively.

Expression of immediate-early genes in the dorsal cochlear nucleus in salicylate-induced tinnitus.

Spontaneous neuronal activity in dorsal cochlear nucleus (DCN) may be involved in the physiological processes underlying salicylate-induced tinnitus. As a neuronal activity marker, immediate-early gene (IEG) expression, especially activity-dependent cytoskeletal protein (Arc/Arg3.1) and the early growth response gene-1 (Egr-1), appears to be highly correlated with sensory-evoked neuronal activity. However, their relationships with tinnitus induced by salicylate have rarely been reported in the DCN. In this study, we assessed the effect of acute and chronic salicylate treatment on the expression of N-methyl D-aspartate receptor subunit 2B (NR2B), Arg3.1, and Egr-1. We also observed ultrastructural alterations in the DCN synapses in an animal model of tinnitus. Levels of mRNA and protein expression of NR2B and Arg3.1 were increased in rats that were chronically administered salicylate (200 mg/kg, twice daily for 3, 7, or 14 days). These levels returned to baseline 14 days after cessation of treatment. However, no significant changes were observed in Egr-1 gene expression in any groups. Furthermore, rats subjected to long-term salicylate administration showed more presynaptic vesicles, thicker and longer postsynaptic densities, and increased synaptic interface curvature. Alterations of Arg3.1 and NR2B may be responsible for the changes in the synaptic ultrastructure. These changes confirm that salicylate can cause neural plasticity changes at the DCN level.

[Proteomic analysis of gastric mucosa in chronic gastritis rats of Pi-Wei damp-heat syndrome treated by sanren decoction: an experimental study].

OBJECTIVE: To study the expressions of gastric mucosal proteins in chronic gastritis (CG) rats of Pi-Wei damp-heat syndrome (PWDHS), to investigate the pathogenesis correlated to CG rats of PWDHS, to observe the differential expressions of gastric mucosal proteins in CG rats of PWDHS, and to investigate the mechanisms of Sanren Decoction (SD) for treating CG rats of PWDHS. METHODS: Totally 36 male SD rats were adaptable fed for 3 days and randomly divided into 3 groups, i.e., the normal control group, the CG of PWDHS rat model group (abbreviated as the model group), and the SD treatment group, 12 in each group. The CG of PWDHS rat model was prepared by composite factors. The gastric mucosal protein was separated using two-dimensional gel electrophoresis technique, and stained by Coomassie brilliant blue. The protein spots expressed differently were analyzed by PDquest 8.0 software. The protein spots expressed differently was identified by MALDI-TOF/TOF-MS. RESULTS: The protein spots were 1 025 +/- 3 9, 994 +/- 51, 1 087 +/- 33 in the normal control group, the model group, and the SD treatment group respectively detected from two-dimensional gel electrophoresis profiles. Compared with the normal control group, there were 74 protein spots differentially expressed in the model group, 30 spots up-regulated and 44 spots down-regulated. Compared with the model group, there were 75 protein spots differentially expressed in the SD treatment group, 49 spots up-regulated and 26 spots down-regulated. Five protein spots differentially expressed were successfully identified, i.e., heat shock protein 72 (HSP72), heat shock protein 60 (HSP60), protein disulfide-isomerase (PDI), malate dehydrogenase (MDH), and unnamed protein. CONCLUSIONS: The pathogenesis of CG of PWDHS may be correlated to energy metabolism disturbance and stress. The mechanisms of SD for treating it may possibly adjust differential expressions of gastric mucosal proteins.

[Double suicide gene therapy with RNAi targeting to STAT3 inhibits the growth of colorectal carcinoma cells in vitro].

OBJECTIVE: The aim of this study was to construct a recombinant plasmid carrying fusion suicide gene CDglyTK and RNA interference eukaryotic expressing vector targeting to STAT3, and to investigate the effect of double suicide gene combined with RNAi targeting to STAT3 on HCT116 and HUVEC cells in vitro. METHODS: The CD and TK were cloned by polymerase chain reaction (PCR), and fusion gene CDglyTK was inserted into plasmid pEGFP after DNA sequence analysis, enzyme digestion and ligation. The recombinant plasmid was analyzed by PCR amplification and electrophoresis and enzyme digestion. DNA sequences containing small hairpin structure targeting to STAT3 were synthesized and inserted into the vector. The CDglyTK gene expressions in HCT116 and HUVEC cells were examined by reverse transcription-polymerase chain reaction (RT-PCR) after transfection of HCT116 and HUVEC cells. The inhibitory effect of RNA interference vector targeting to STAT3 was analyzed by RT-PCR and Western blot. The effects of 5-FC and GCV on HCT116 and HUVEC cells transfected with the recombinant plasmids were detected by MTT staining. RESULTS: The results of restriction enzyme digestion and PCR amplification and electrophoresis showed that the recombinant pEGFP/CDglyTK was constructed correctly. The mRNA expression of gene CDglyTK was detected in HCT116 and HUVEC cells which transfected with the recombinant plasmid. The results of RT-PCR and Western blot showed that the RNA interference expression vector targeting to STAT3 effectively inhibited the expression of STAT3 in HCT116 cells. The results of MTT test showed that the inhibition ratio of group pEGFP/CDglyTK was (63.72 ± 0.64)%, significantly higher than that of control group (P < 0.05). The inhibition rate of group pEGFP/STAT3 siRNA was (47.02 ± 0.39)%, which was lower than that of group pEGFP/CDglyTK (P < 0.05), and higher than that of control group (P < 0.05). The inhibition rate of group pEGFP/CdglyTK + pEGFP/STAT3 siRNA was (85.10 ± 0.17)%, significantly higher than those of groups pEGFP/CDglyTK and group pEGFP/STAT3 siRNA (P < 0.05). Meanwhile, in HUVEC cells, the inhibition rate of group pEGFP/CDglyTK was (70.24 ± 0.33)%, significantly higher than that of the control group (P < 0.05). The inhibition rate of group pEGFP/STAT3 siRNA was (46.32 ± 0.15)%, significantly lower than that of group pEGFP/CdglyTK (P < 0.05), and higher than that of the control group (P < 0.05). The inhibition rate of group pEGFP/CdglyTK+pEGFP/STAT3 siRNA was (87.10 ± 0.24)%, significantly higher than those of groups pEGFP/CDglyTK and pEGFP/STAT3 siRNA(P < 0.05). CONCLUSION: The recombinant plasmids pEGFP-CDglyTK and pEGFP/STAT3 siRNA have inhibitory effect on HCT116 and HUVEC cells. The killing effects of double suicide gene combined with RNAi targeting to STAT3 are much better than those of single gene therapy.

Inflammation and DNA methylation coregulate the CtBP-PCAF-c-MYC transcriptional complex to activate the expression of a long non-coding RNA CASC2 in acute pancreatitis.

Long non-coding RNAs (lncRNAs) are emerging as important regulators involved in the pathogenesis of many diseases. However, it is still unknown if they contribute to the occurrence of acute pancreatitis (AP). Here, we identified a lncRNA CASC2 (Cancer Susceptibility Candidate 2) was significantly upregulated in the pancreatic tissues from AP patients. Knockdown or overexpression of CASC2 in vitro could specifically repress or induce the expression of two proinflammatory cytokines including IL6 (Interleukin 6) and IL17, respectively. Changing the expression levels of several transcription factors that were predicted to bind to the promoter of CASC2, we found c-MYC could specifically regulate the expression of CASC2. Using immunoprecipitation, mass spectrometry, and co-immunoprecipitation assays, we proved that c-MYC assembled a transcriptional complex with PCAF (p300/CBP-associated Factor) and CtBP1/2 (C-terminal Binding Protein 1 and 2), terming as the CtBP-PCAF-c-MYC (CPM) complex. Further investigation revealed that CtBPs were amplified in the pancreatic tissues from AP patients and they functioned as coactivators to induce the expression of CASC2 and thus led to the upregulation of IL6 and IL17. Moreover, we identified that decreased DNA methylation levels in the promoters of CtBPs and inflammatory stimuli coactivated the expression of CtBPs. Collectively, we identified a new signaling pathway in which DNA methylation and inflammatory stimuli coregulate the CPM complex to activate CASC2 expression, whose induction further activates the expression of IL6 and IL17, eventually aggravating inflammation response and causing the pathology of AP.

Septin 9 Methylation in Nasopharyngeal Swabs: A Potential Minimally Invasive Biomarker for the Early Detection of Nasopharyngeal Carcinoma.

Nasopharyngeal carcinoma (NPC) is highly prevalent in Southeast Asia, and an unfavorable outcome is usually attributed to advanced stage NPC. Current methods for the early diagnosis of NPC have limitations in clinical practice. The aim of this study was to investigate the diagnostic ability of Septin 9 methylation for NPC. A quantitative methylation-sensitive PCR (qMS-PCR) assay was developed to measure the methylation status and levels of Septin 9 in nasopharyngeal tissues and paired swabs from patients with NPC, chronic nasopharyngitis, and healthy donors. Methylated Septin 9 was detected in 92% (23/25) of NPC tissues and 25% (4/16) of nasopharyngitis controls (p < 0.05). High-frequency hypermethylation with decreased mRNA expression of Septin 9 in NPC was also identified. Further, Septin 9 methylation was identified in 90.5% (19/21) of NPC biopsies and 71.4% (15/21) of paired swabs, indicating a good concordance between the two sample types. In addition, methylated Septin 9 was found in 16 (72.7%) nasal swabs from 22 NPC patients, 2 of 19 (10.5%) nasopharyngitis, but not in any of the healthy controls (p < 0.01). The methylation score in nasal swabs of the NPC group was also significantly higher than that of non-NPC controls (p < 0.001). Moreover, receiver operating characteristic (ROC) curve analysis showed an area under the curve (AUC) of 0.882 of Septin 9 methylation tests to discriminate NPC from non-NPC subjects. Our study demonstrated that frequent methylation of Septin 9 was present in NPC. Its detection in nasopharyngeal swabs may provide a minimally invasive and informative method for identifying early NPC cases.

[Rhubarb induced change of tumor necrosis factor-alpha level in guinea pig model of melanosis coli and its significance].

OBJECTIVE: To study the induced change of tumor necrosis factor alpha (TNF-alpha) level in serum and colon tissue of guinea pig model with melanosis coli and its significance induced by rhubarb (RB). METHODS: One hundred and twelve guinea pigs of clean grade were randomly divided into four groups: the 16 in the normal group (untreated) and the 3 RB groups (32 in each) treated with low (3 g/kg d), medium (6 g/kg d), and high (12 g/kg d) dose of rhubarb respectively, administered by gastrogavage for 60 successive days. All guinea pigs were sacrificed at the terminal of the experiment and their blood serum and colon tissue were taken for detecting TNF-alpha level and TNF-alpha mRNA expression qualitatively and quantitatively using ELISA and RT-PCR. RESULTS: Compared with the normal group, serum and colonic tissue levels of TNF-alpha and TNF-alpha mRNA expression in the RB groups were higher significantly (P<0.01), while no significant difference was found among the later three groups (P>0.05). CONCLUSION: RB could induce change of TNF-alpha level in serum and colon tissue of guinea pig with melanosis coli.

The Polymorphism rs17525495 of LTA4H Is Associated with Susceptibility of Crohn's Disease instead of Intestinal Tuberculosis in a Chinese Han Population.

BACKGROUND: Because of the similarity of intestinal tuberculosis and Crohn's disease in disease phenotype, differential diagnosis has always been a clinical problem. Arachidonic acid metabolites play an important role in the inflammatory response of intestinal tuberculosis and Crohn's disease. Recent studies have shown that the polymorphism locus in the promoter region of LTA4H gene affects LTB4 expression level and the susceptibility to extrapulmonary tuberculosis. Thus, we identified a total of 148 patients with intestinal tuberculosis, 145 with Crohn's disease, and 700 normal controls in this study. METHODS: All the study participants were local Han people from Jiangxi Province in the past eleven years. DNA was extracted from the paraffin-embedded specimens or the whole blood. The LTA4H promoter SNP (rs17525495) was genotyped with TaqMan assay. RESULTS: The T-alleles frequency was not significantly increased in patients with intestinal tuberculosis compared with healthy control group (p=0.630; OR=1.07; 95%CI=0.81-1.41), while patients with Crohn's disease have significantly increased T allele frequency compared with healthy population (p=0.032; OR=1.34; 95%CI=1.03-1.75). During treatment, the presence of the T allele significantly increased the proportion of Crohn's patients requiring glucocorticoids (p<0.05). CONCLUSIONS: The T allele of LTA4H gene SNP (rs17525495) is a risk factor for Crohn's disease instead of intestinal tuberculosis. More importantly, there may be a potential association of the different genotypes of rs17525495 with the treatment efficacy of 5-ASA and glucocorticoids in patients with Crohn's disease. The association between LTA4H polymorphism and drugs therapeutic effects might contribute to the practice of precision medicine and the prediction of clinical outcomes.

Interleukin-22 receptor 1 is expressed in multinucleated giant cells: A study on intestinal tuberculosis and Crohn's disease.

BACKGROUND: It is challenging to distinguish intestinal tuberculosis from Crohn's disease due to dynamic changes in epidemiology and similar clinical characteristics. Recent studies have shown that polymorphisms in genes involved in the interleukin (IL)-23/IL-17 axis may affect intestinal mucosal immunity by affecting the differentiation of Th17 cells. AIM: To investigate the specific single-nucleotide polymorphisms (SNPs) in genes involved in the IL-23/IL-17 axis and possible pathways that affect susceptibility to intestinal tuberculosis and Crohn's disease. METHODS: We analysed 133 patients with intestinal tuberculosis, 128 with Crohn's disease, and 500 normal controls. DNA was extracted from paraffin-embedded specimens or whole blood. Four SNPs in the IL23/Th17 axis (IL22 rs2227473, IL1ß rs1143627, TGFß rs4803455, and IL17 rs8193036) were genotyped with TaqMan assays. The transcriptional activity levels of different genotypes of rs2227473 were detected by dual luciferase reporter gene assay. The expression of IL-22R1 in different intestinal diseases was detected by immunohistochemistry. RESULTS: The A allele frequency of rs2227473 (P = 0.030, odds ratio = 0.60, 95% confidence interval: 0.37-0.95) showed an abnormal distribution between intestinal tuberculosis and healthy controls. The presence of the A allele was associated with a higher IL-22 transcriptional activity (P < 0.05). In addition, IL-22R1 was expressed in intestinal lymphoid tissues, especially under conditions of intestinal tuberculosis, and highly expressed in macrophage-derived Langhans giant cells. The results of immunohistochemistry showed that the expression of IL-22R1 in patients with Crohn's disease and intestinal tuberculosis was significantly higher than that in patients with intestinal polyps and colon cancer (P < 0.01). CONCLUSION: High IL-22 expression seems to be a protective factor for intestinal tuberculosis. IL-22R1 is expressed in Langhans giant cells, suggesting that the IL-22/IL-22R1 system links adaptive and innate immunity.

Preparation and characterization of novel nanocomposite films formed from silk fibroin and nano-TiO2.

This paper describes the synthesis and characterization of new regenerated silk fibroin (SF)/nano-TiO(2) composite films. The preparation method, based on the sol-gel technique using butyl titanate as oxide precursor, could avoid reagglomeration of the prepared nanoparticles. Samples were characterized mainly by X-ray diffraction (XRD), ultra-violet (UV) spectroscopy, atomic force microscopy (AFM), Fourier transform infrared (FT-IR) spectroscopy, and thermogravimetric analysis (TGA). The UV and AFM results indicated that TiO(2) nanoparticles could be well dispersed inside the SF film, and the size of TiO(2) was about 80nm. The XRD and FT-IR analysis implied that the formation of nano-TiO(2) particles may induce the conformational transition of silk fibroin to a typical Silk II structure partly with the increasing of crystallinity in the composite films. Compared to the pure SF films, the mechanical and thermal properties of composite films were improved, and the solubility in water was decreased due to the conformational transition of silk fibroin to Silk II structure.

[Protective role of Pim-3 gene in intestinal mucosa damaged by burn or lipopolysaccharide].

OBJECTIVE: To explore the protective role of Pim-3 gene in intestinal mucosa damaged by burn or lipopolysaccharide (LPS). METHODS: (1) Ninety Wistar mice were randomly divided into 3 equal groups: Group A, undergoing 30% grade III burning on the back; Group B, undergoing intraperitoneal injection of LPS; and Group C, undergoing intraperitoneal injection of normal saline. 3, 6, 12, 24, and 48 hours after the treatment 5 rats from each group were killed with their small intestine tissues taken out. RT-PCR was used to detect the mRNA expression of Pim-3, a serine/threonine kinase, occludin, intercellular adhesion molecule (ICAM)-1, and Western blotting was used to detect the protein expression of Pim-3 and occludin. (2) Intestinal endothelial cells (IECs) of newborn Wistar rats were collected, cultured, and divided into 6 groups: Group (1), treated with LPS (endotoxin), Group (2), transfected with blank plasmid pEGFP-N(2) and treated with LPS, Group (3), transfected with recombinant pEGFP-N(2)/Pim-3and treated with LPS, Group (4), as normal control group, Group (5), transfected with blank plasmid pEGFP-N(2), and Group (6), transfected with recombinant plasmid pEGFP-N(2)/Pim-3. Six hours later RT-PCR was used to detect the mRNA expression of Pim-3, ICAM-1, and occludin. The apoptosis of the cells was examined by flow cytometry. RESULTS: (1) In Groups A and B the mRNA expression of Pim-3 began to increase 3 h later, peaked 6 h later, and then gradually decreased. The Pim-3 mRNA expression of Group C, however, remained always at a low level. The ICAM-1 mRNA expression levels of Groups A and B were constantly up-regulated 6 h later, all significantly higher than those of Group C (all P < 0.01). The occludin mRNA expression levels of Groups A and B began to increase 3 hours later, and peaked 12 hours later, all significantly higher than those of Group C (all P < 0.05). (2) The mRNA expression levels of Pim-3 and occludin of Group (6) was significantly higher than those of Groups (4) and (5) (both P < 0.05). However, there was no significant difference in ICAM-1 mRNA expression among Groups (4), (5), and (6). The mRNA expression levels of Pim-3, occluding, and ICAM-1 of Groups (3) were all significantly higher than those of Groups (1) and (2) (all P < 0.05). The mRNA expression levels of ICAM-1 of Groups (1), (2), and (3) were all significantly lower than those of Groups (4), (5), and (6) (all P < 0.05). The apoptotic rates of groups (4), (5), and (6) were all very low. The apoptotic rates of Groups (1) and (2) were 41.3% and 44.80% respectively, both significantly higher than that of Group (3) (36.03%, both P < 0.05). CONCLUSION: Both burning and LPS stimulate endogenous Pim-3 gene expression in small intestine which lasts a short time. Pim-3 gene not only suppresses the apoptosis of IECs, but also strengthens the occludin expression and inhibits the intestinal tract inflammatory reaction induced by LPS. LPS induces ICAM-1 expression, which can be inhibited by Pim-3.

Effect of drug treatment on hyperplastic gastric polyps infected with Helicobacter pylori: a randomized, controlled trial.

AIM: To study the effects of drug treatment on hyperplastic gastric polyps infected with Helicobacter pylori (H pylori). METHODS: Forty-eight patients with hyperplastic gastric polyps (3-10 mm in diameter) infected with H pylori were randomly assigned to a treatment group (n = 24) which received proton-pump inhibitor (omeprazole or lansoprazole), clarithromycin, bismuth citrate and tinidazole, and a control group (n = 24) which received protective agent of gastric mucosa (tepretone). Patients underwent endoscopy and H pylori examination regularly before enrollment and 1-12 mo after treatment. RESULTS: Twenty-two patients in the treatment group and 21 in the control group completed the entire test protocol. In the treatment group, polyps disappeared 1-12 mo (average, 6.5+/-1.1 mo) after the treatment in 15 of 22 patients (68.2%) and H pylori infection was eradicated in 19 of the 22 patients (86.4%). However, 12 months after the study, no change in polyps or H pylori status was seen in any controls ((b)P<0.01). CONCLUSION: Most hyperplastic gastric polyps disappear after eradication of H pylori.

[Comparative study on Jinghua Weikang Capsule and famotidine in treating duodenal ulcer].

OBJECTIVE: To compare the effect and security of Jinghua Weikang Capsule (JWC) and famotidine in treating duodenal ulcer. METHODS: Two hundred patients with duodenal ulcer were randomly divided into the treated group treated with JWC, 160 mg 3 times per day and the control group treated with famotidine 20 mg twice per day, both by orally taking before meal for 4 weeks, 100 cases in each group. Changes of symptom score, adverse reaction, helicobacter pylori (Hp) infection and endoscopic figure before and after treatment were observed. RESULTS: The general remission rate and the ulcer healing rate in the treated group were equal to those in the control group respectively (both P > 0.05). Symptoms including belly ache, sour regurgitation and abdominal distension were ameliorated after therapy in both groups (all P < 0.05). The effect in relieving anorexia and eructation and Hp eradication rate were significantly higher in the treated group than those in the control group (all P< 0.05), while the incidence rate of adverse reaction in the treated group was remarkably lower than that in the control group (P < 0.05). CONCLUSION: JWC is an effective and safe remedy in treating duodenal ulcer, especially for symptom amelioration and Hp eradication, so it is worthy of expanding clinically.

[Effects of acupuncture on the gastric motivity in patients with functional dyspepsia].

OBJECTIVE: To observe the effects of the acupuncture on the gastric motivity in patients with functional dyspepsia (FD). METHODS: Ninety FD patients were randomly divided into 3 groups: the treated group, the control groups, 1 and 2, 30 patients in each group. Patients in the treated group received acupuncture treatment, those in the control group 1 and group 2 orally took Cisapride and Marzulene-s granule respectively. Changes in scores of symptoms, electrogastrographic figure, gastric emptying time estimated by B-ultrasonic examination and plasma level of motilin in the 3 groups before and after treatment were observed. RESULTS: Symptoms were obviously improved in the treated and control group 1 after treatment, which were better than those in the control group 2 (P < 0.01). Gastro-electric frequency and rhythm in the treated group and control group 1 were improved more significantly than those in before treatment (P < 0.01). Gastric emptying time and plasma motilin improved in the treated group and the control group 1 after treatment, when compared with before treatment, the difference was significant respectively (P < 0.05). CONCLUSION: Acupuncture could significantly improve the gastric motivity of FD patients.

Overexpression of pim-3 and protective role in lipopolysaccharide-stimulated hepatic stellate cells.

AIM: To investigate pim-3 expression in hepatic stellate cells (HSCs) stimulated by lipopolysaccharide (LPS), and its protective effect on HSCs. METHODS: Rat HSC-T6 cells were stimulated by LPS. The effect of LPS on proliferation and apoptosis of HSC-T6 cells was investigated by methyl thiazoyltetrazolium (MTT) assay and flow cytometry after annexin V-fluorescein isothiocyanate/propidium iodide double staining. pim-3 mRNA and protein were detected by reverse transcriptase polymerase chain reaction and Western blotting at 48 h when HSC-T6 cells were stimulated with 1 µg/mL LPS for 0, 3, 6, 12, 24 and 48 h. The cells without stimulation served as controls. To study the effect of pim-3 kinase on HSC-T6 cells, si-pim3 (siRNA against pim-3) was transfected into HSC-T6 cells. HSC-T6 cells were subjected to different treatments, including LPS, si-pim3, or si-pim3 plus LPS, and control cells were untreated. Protein expression of pim-3 was detected at 48 h after treatment, and cell proliferation at 24 and 48 h by MTT assay. Apoptosis was detected by flow cytometry, and confirmed with caspase-3 activity assay. RESULTS: LPS promoted HSC-T6 cell proliferation and protected against apoptosis. Significantly delayed upregulation of pim-3 expression induced by LPS occurred at 24 and 48 h for mRNA expression (pim-3/ß-actin RNA, 24 or 48 h vs 0 h, 0.81 ± 0.20 or 0.78 ± 0.21 vs 0.42 ± 0.13, P < 0.05), and occurred at 12 h and peaked at 24 and 48 h for protein expression (pim-3/GAPDH protein, 12, or 24 or 48 h vs 0 h, 0.68 ± 0.12, 1.47 ± 0.25 or 1.51 ± 0.23 vs 0.34 ± 0.04, P < 0.01). pim-3 protein was ablated by si-pim3 and upregulated by LPS in HSC-T6 cells at 48 h after treatment (pim-3/GAPDH: si-pim3, si-pim3 plus LPS or LPS vs control, 0.11 ± 0.05, 0.12 ± 0.05 or 1.08 ± 0.02 vs 0.39 ± 0.03, P < 0.01). Ablation of pim-3 by si-pim3 in HSC-T6 cells partly abolished proliferation (OD at 24 h, si-pim3 group or si-pim3 plus LPS vs control, 0.2987 ± 0.050 or 0.4063 ± 0.051 vs 0.5267 ± 0.030, P < 0.01; at 48 h 0.4634 ± 0.056 or 0.5433 ± 0.031 vs 0.8435 ± 0.028, P < 0.01; si-pim3 group vs si-pim3 plus LPS, P < 0.01 at 24 h and P < 0.05 at 48 h), and overexpression of pim-3 in the LPS group increased cell proliferation (OD: LPS vs control, at 24 h, 0.7435 ± 0.028 vs 0.5267 ± 0.030, P < 0.01; at 48 h, 1.2136 ± 0.048 vs 0.8435 ± 0.028, P < 0.01). Ablation of pim3 with si-pim3 in HSC-T6 cells aggravated apoptosis (si-pim3 or si-pim3 plus LPS vs control, 42.3% ± 1.1% or 40.6% ± 1.3% vs 16.8% ± 3.3%, P < 0.01; si-pim3 vs si-pim3 plus LPS, P > 0.05), and overexpression of pim-3 in the LPS group attenuated apoptosis (LPS vs control, 7.32% ± 2.1% vs 16.8% ± 3.3%, P < 0.05). These results were confirmed by caspase-3 activity assay. CONCLUSION: Overexpression of pim-3 plays a protective role in LPS-stimulated HSC-T6 cells.

Alkaloids from Menispermum dauricum.

The alkaloids, dechloroacutumidine and 1-epidechloroacutumine, together with three known alkaloids, acutumidine, acutumine, and dechloroacutumine, were isolated from the rhizomes of Menispermum dauricum and their structures established by spectral and chemical methods. The cytotoxicity of each compound against the growth of human cell lines was studied, and acutumine selectively inhibited T-cell growth.

Serotonin excites arcuate neurons directly but inhibits them through intercalated GABAergic neurons.

Effects of serotonin (5-HT) on spontaneous discharges of single hypothalamic arcuate neurons were observed in rat brain slices by extracellular recordings. The results showed that (1) of 149 neurons selected randomly and tested for 5-HT application, 33 (22.2%) were excited, 82 (55.0%) were inhibited, and 34 (22.8%) showed biphasic responses or failed to respond; (2) substitution of low Ca(2+)-high Mg(2+) artificial cerebrospinal fluid (aCSF) for normal aCSF abolished the 5-HT-induced inhibitory effect but failed to affect the 5-HT-induced excitatory effect; (3) cyproheptadine, a non-selective 5-HT receptor antagonist, could block either the 5-HT-induced excitatory or inhibitory effects in all neurons tested; and (4) bicuculline, a GABA(A)-receptor antagonist, blocked the 5-HT-induced inhibitory effect. These results imply (1) 5-HT excites arcuate neurons through a mechanism that is insensitive to the decreased extracellular Ca(2+), suggesting a direct postsynaptic action of 5-HT on the 5-HT-receptors located in the membrane of the neurons recorded; and (2) 5-HT might elicit the inhibitory effect through a Ca(2+)-sensitive release of GABA from intercalated GABAergic local neurons that are excited first by 5-HT.