Luteolin promotes neuronogenesis in hippocampus of chronic unpredictable mild stress rats and primary hippocampus of fetal rats.

OBJECTIVE: To investigate the effects of luteolin on chronic unpredictable mild stress (CUMS)-induced depressive rats and corticosterone (CORT)-induced depressive primary hippocampal neurons, and to elucidate the mechanism behind the action. METHODS: The antidepressant mechanism of luteolin was studied by using CUMS rat model and primary hippocampal neurons in fetal rats. In vivo, novelty suppressed feeding, open-field and sucrose preference tests as well as Morris water maze were evaluated. The content of brain derived neurotrophic factor (BDNF), 5-hydroxytryptamine (5-HT), norepinephrine (NE), and dopamine (DA) in serum were detected by enzyme-linked immunosorbent assay. The mechanisms of luteolin were explored based on neurotrophin and hippocampal neurogenesis, and proliferation. Survival of the septo-temporal axis in hippocampus was assayed using the 5-bromo-2-deoxyuridine (BrdU), the expression of BDNF, neurotrophin-3 (NT-3), and nerve growth factor (NGF) in hippocampus dentate gyrus region were measured by Western-blotting. In vitro, BDNF, NT-3, tropomyosin receptor kinase B (TrkB), and phosphorylated cyclic adenosine monophosphate responsive element binding protein (p-CREB) were detected through the high content analysis (HCA) to investigate neurotrophin and apoptosis. RESULTS: Induction of CUMS in rats induced depressive symptoms, while luteolin significantly enhanced sucrose consumption, decreased feeding latency, increased locomotor activity, escape latency, distance of target quadrant and regulated the content of depressive-like biomarkers. Histology analysis revealed that luteolin increased the abundance of new born neurons that had been labeled with BrdU, BrdU + neuronal nuclear antigen, and BrdU + doublecortin in septo-temporal axis of S2 (mid-septal) and T3 (mid-temporal). Moreover, expression of BDNF, NT-3, and NGF increased significantly in the septo-temporal axis of S2 and T3. HCA showed increased expression of BDNF, NT-3, TrkB and p-CREB in primary hippocampal neurons. CONCLUSION: The results provided direct evidence that luteolin has an antidepressant effect and could effectively promote the regeneration of the septotemporal axis nerve and hippocampal neuronutrition, which suggested that the antidepressant effect of luteolin may be related to hippocampal neurogenesis.

Effects of Jianpi Yichang Powder on Expressions of IL-1β and IL-18 of NLRP3 Signaling Pathway in Ulcerative Colitis Rats / 中国中医药信息杂志

Objective To explore the effects of Jianpi Yichang Powder on the expressions of interleukin(IL)-1β and IL-18 of NLRP3 signaling pathway in ulcerative colitis(UC)model rats.Methods Ten rats were randomly selected from 40 SD rats as the normal group,and the other rats freely drank 5%dextran sulfate solution for 7 days to replicate UC rats model.The model rats were randomly divided into model group,sulfasalazine group and Jianpi Yichang Powder group,with 10 rats in each group.Jianpi Yichang Powder group and sulfasalazine group were given corresponding liquid medicine for gavage,and the normal and model groups were given equivalent volume distilled water for gavage for consecutive 14 d.The general status was observed,and the disease activity index(DAI)was scored,the contents of NLRP3,apoptosis-associated spotted proteins(ASC),and Caspase-1 in serum were detected by ELISA,the expressions of IL-1β and IL-18 protein and mRNA in colon tissue were detected by immunohistochemistry,Western blot and RT-PCR respectively.Results Compared with the normal group,the general status of the rats in model group was relatively worse,and DAI score significantly increased(P<0.01),the contents of NLRP3,ASC and Caspase-l in serum were significantly increased(P<0.01),the expressions of IL-1β and IL-18 protein and mRNA in colon tissue were significantly increased(P<0.01).Compared with the model group,the general status of the rats in Jianpi Yichang Powder group and sulfasalazine group were significantly improved,DAI score significantly decreased(P<0.01),the contents of NLRP3,ASC and Caspase-l in serum significantly reduced(P<0.05,P<0.01),and the expressions of IL-1β and IL-18 protein and mRNA in colon tissue significantly decreased(P<0.05,P<0.01).Conclusion Jianpi Yichang Powder can inhibit IL-1β and IL-18 expression of NLRP3 signaling pathway to reduce colon immune inflammatory damage,thus play a role in treating UC.

Effects of Regulation of SHH/Gli1 Signaling Pathway by Baishile Capsules on Hippocampal Neurogenesis Depression Model Rats / 中国中医药信息杂志

Objective To explore the effects and mechanism of Baishile Capsules regulating SHH/Gli1 signaling pathway on hippocampal neurogenesis of depression model rats.Methods Totally 32 SD rats were randomly divided into control group,model group,fluoxetine(5.4 mg/kg)group and Baishile Capsules(2.88 g/kg)group,with 8 rats in each group.A depression rat model was established using chronic unpredictable mild stress and single cage feeding method.The model was established and administered simultaneously for 21 consecutive days.Depression-like behavior in rats were evaluated by sucrose preference experiment and open field experiment,ELISA was used to detect brain derived neurotrophic factor(BDNF)contents in rat serum and hippocampal tissue,the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus was observed by immunofluorescence,immunofluorescence and Western blot were used to detect the fluorescence intensity and protein expression of SHH,Gli1,Smo,Ptch in hippocampal tissue.Results Compared with the control group,the degree of sucrose preference significantly decreased in the model group(P<0.01),the number of horizontal and vertical movements significantly decreased(P<0.01),the contents of BDNF in serum and hippocampal tissue significantly decreased(P<0.05),the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus significantly decreased(P<0.01),and the fluorescence intensity and protein expression of SHH,Gli1,Smo,Ptch in hippocampal tissue significantly decreased(P<0.01,P<0.05).Compared with the model group,the degree of sucrose preference and the number of horizontal and vertical movements in fluoxetine group and Baishile Capsule group increased significantly(P<0.05,P<0.01),the contents of BDNF in serum and hippocampal tissue significantly increased(P<0.05,P<0.01),and the number of BrdU,BrdU/DCX,BrdU/NeuN positive cells in dentate gyrus of the hippocampus significantly increased(P<0.01,P<0.05),the fluorescence intensity and protein expressions of SHH,Gli1,Smo,Ptch in hippocampal tissue significantly increased(P<0.01,P<0.05).Conclusion Baishile Capsule can promote the hippocampus neurogenesis in depression model rats by regulating SHH/Gli1 signaling pathway,and play an antidepressant role.

Research Progress in the Treatment of Ulcerative Colitis with Sishen Pills / 中国中医药信息杂志

Sishen Pills is a classic prescription for the treatment of spleen and kidney diarrhea,which has the effect of warming the kidney and the spleen,astringent intestine and antidiarrheal.In modern clinical application,the modified prescriptions based on Sishen Pills,combined with other treatments of TCM and integrated traditional Chinese and Western medicine are often used to treat ulcerative colitis with spleen-kidney yang deficiency syndrome,and the curative effect is remarkable.Experimental pharmacological studies have shown that Sishen Pills may achieve the purpose of ulcerative colitis by regulating the expression of related signaling pathway proteins,regulating inflammatory factors,inhibiting inflammatory response,regulating autophagy,regulating intestinal flora,improving intestinal mucosal permeability,repairing intestinal mucosal barrier,regulating cellular energy metabolism,anti-oxidative stress,regulating cellular immune function,etc.In this article,the research status of Sishen Pills in the treatment of ulcerative colitis was sorted out and summarized,in order to provide reference for further study of its mechanism and clinical application.

Effect of Jianpi Yichang Powder on NLRP3/ASC/Caspase-l Signaling Pathway in Rats with Ulcerative Colitis / 中国实验方剂学杂志

ObjectiveTo explore the effect of Jianpi Yichang power on the nucleotide-binding oligomerization domain-like receptor protein 3 （NLRP3） inflammasome signaling pathway in a rat model of ulcerative colitis （UC）. MethodSixty Sprague-Dawley rats were randomly divided into a normal group （n=10） and an experimental group （n=50）. The experimental group received 5% dextran sulfate sodium （DSS） solution freely for 7 days to induce UC， and then they were further randomly divided into model group， sulfasalazine （0.3 g·kg-1） group， and high-， medium-， and low-dose Jianpi Yichang power groups （54.4， 27.2， 13.6 g·kg-1） for continuous treatment of 14 days. The general condition of the rats was observed and recorded daily， and the disease activity index （DAI） was scored before and after treatment. Enzyme-linked immunosorbent assay （ELISA） was used to detect the levels of interleukin-1β （IL-1β） and interleukin-18 （IL-18） in the serum of rats in each group. Hematoxylin-eosin （HE） staining was performed to observe the histopathological changes in the colon tissue. Immunohistochemistry， Western blot， and Real-time polymerase chain reaction （Real-time PCR） were used to detect the positive protein expression， protein expression， and mRNA expression of NLRP3， apoptosis-associated speck-like protein containing a CARD （ASC）， and cysteine aspartate-special proteases-1（Caspase-1） in the colon tissue. ResultCompared with the condition in the normal group， the general condition of rats in the model group was relatively poor， with increased DAI scores （P<0.01）， pathological changes in the colon， increased levels of IL-1β and IL-18 in the serum （P<0.01）， and enhanced positive protein expression， protein expression， and mRNA expression of NLRP3， ASC， and Caspase-1 in the colon tissue （P<0.01）. Compared with the condition in the model group， the general condition of rats in the Jianpi Yichang power groups at various doses improved significantly， with reduced DAI scores （P<0.05， P<0.01）， alleviated pathological changes in the colon as revealed by HE staining， and reduced protein expression levels of NLRP3 and Caspase-1 in the colon tissue （P<0.05， P<0.01）. The serum levels of IL-1β and IL-18， and ASC protein expression in the colon， as well as the mRNA expression levels of NLRP3， ASC， and Caspase-1， decreased in the high- and medium-dose Jianpi Yichang power groups （P<0.05， P<0.01）. The positive protein expression levels of NLRP3， ASC， and Caspase-1 were reduced in the high-dose Jianpi Yichang power group （P<0.01）. The positive protein expression levels of ASC and Caspase-1 were reduced in the medium-dose Jianpi Yichang power group （P<0.05）. The mRNA expression level of ASC was reduced in the low-dose Jianpi Yichang power group （P<0.05）. ConclusionJianpi Yichang power can reduce colon immune inflammatory damage by regulating the NLRP3 inflammasome signaling pathway， thereby exerting a role in treating UC.

Nucleocapsid 203 mutations enhance SARS-CoV-2 immune evasion

Previous work indicated that the nucleocapsid 203 mutation increase the virulence and transmission of the SARS-CoV-2 Alpha variant. However, Delta later outcompeted Alpha and other lineages, promoting a new wave of infections. Delta also possesses a nucleocapsid 203 mutation, R203M. Large-scale epidemiological analyses suggest a synergistic effect of the 203 mutation and the spike L452R mutation, associated with Delta expansion. Viral competition experiments demonstrate the synergistic effect in fitness and infectivity. More importantly, we found that the combination of R203M and L452R brings in a 3.2-fold decrease in neutralizing titers to the neutralizing serum relative to L452R-only virus. R203M/L452R show an increased fitness after the initiation of global vaccination programmes, possibly associated with the enhanced immune evasion. Another rapidly emerging variant Omicron also bears the 203 mutation. Thus, we proposed that nucleocapsid mutations play an essential role for the rise and predominance of variants in concern.

Unconventional secretion of unglycosylated ORF8 is critical for the cytokine storm during SARS-CoV-2 infection

Coronavirus disease 2019 is a respiratory infectious disease caused by the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Evidence on the pathogenesis of SARS-CoV-2 is accumulating rapidly. In addition to structural proteins such as Spike and Envelope, the functional roles of non-structural and accessory proteins in regulating viral life cycle and host immune responses remain to be understood. Here, we show that open reading frame 8 (ORF8) acts as messenger for inter-cellular communication between alveolar epithelial cells and macrophages during SARS-CoV-2 infection. Mechanistically, ORF8 is a secretory protein that can be secreted by infected epithelial cells via both conventional and unconventional secretory pathways. The unconventionally secreted ORF8 recognizes the IL17RA receptor of macrophages and induces cytokine release. However, conventionally secreted ORF8 cannot bind to IL17RA due to N-linked glycosylation. Furthermore, we found that Yip1 interacting factor homolog B (YIF1B) is a channel protein that translocates unglycosylated ORF8 into vesicles for unconventional secretion. Blocking the unconventional secretion of ORF8 via a YIF1B knockout in hACE2 mice attenuates inflammation and yields delayed mortality following SARS-CoV-2 challenge.

Nucleocapsid mutation R203K/G204R increases the infectivity, fitness and virulence of SARS-CoV-2

In addition to the mutations on the spike protein (S), co-occurring mutations on nucleocapsid (N) protein are also emerging in SARS-CoV-2 world widely. Mutations R203K/G204R on N, carried by high transmissibility SARS-CoV-2 lineages including B.1.1.7 and P.1, has a rapid spread in the pandemic during the past year. In this study, we performed comprehensive population genomic analyses and virology experiment concerning on the evolution, causation and virology consequence of R203K/G204R mutations. The global incidence frequency (IF) of 203K/204R has rose up from nearly zero to 76% to date with a shrinking from August to November in 2020 but bounced later. Our results show that the emergence of B.1.1.7 is associated with the second growth of R203K/G204R mutants. We identified positive selection evidences that support the adaptiveness of 203K/204R variants. The R203K/G204R mutant virus was created and compared with the native virus. The virus competition experiments show that 203K/204R variants possess a replication advantage over the preceding R203/G204 variants, possibly in relation to the ribonucleocapsid (RNP) assemble during the virus replication. Moreover, the 203K/204R virus increased the infectivity in a human lung cell line and induced an enhanced damage to blood vessel of infected hamsters lungs. In consistence, we observed a positive association between the increased severity of COVID-19 and the IF of 203K/204R from in silicon analysis of global clinical and epidemic data. In combination with the informatics and virology experiment, our work suggested the contribution of 203K/204R to the increased transmission and virulence of the SARS-CoV-2. In addition to mutations on the S protein, the mutations on the N protein are also important to virus spread during the pandemic.

Efficacy of angioplasty for tunneled cuffed catheter placement in hemodialysis patients with superior vena cava occlusion / 中华肾脏病杂志

Objective:To evaluate the efficacy of angioplasty on percutaneous superior vena cava occlusion in hemodialysis patients with tunnel-cuffed catheter (TCC) under digital subtraction angiography (DSA) guidance.Methods:A total of 62 hemodialysis patients with TCC in the First Affiliated Hospital of Sun Yat-sen University from December 2017 to June 2020 were enrolled retrospectively. According to the patency of the superior vena cava, the patients were divided into experiment group ( n=20) and control group ( n=42) in this study. Hemodialysis patients with superior vena cava occlusion in the experiment group received angioplasty, including balloon angioplasty, stenting and sharp recanalization, and catheterization with TCC under DSA guidance, while hemodialysis patients without superior vena cava occlusion in the control group only underwent catheterization with TCC under DSA guidance. The 1-year TCC patency rate, postoperative TCC blood flow and treatment-related complications between the two groups were compared. Results:In the experiment group, a total of 11 patients were treated only by percutaneous transluminal angioplasty, while 9 patients were treated combined percutaneous transluminal angioplasty with stent placement. In addition, 3 patients underwent sharp recanalization of superior vena cava occlusion. A total of 9 stents and 29 balloons were used. The course of dialysis in experiment group was longer than that in control group ( P<0.05). There were no significant differences in the 1-year TCC patency rate (85.0% vs 95.2%, P>0.05), postoperative TCC blood flow [(257.83±16.55) ml/min vs (251.90±18.79) ml/min, P>0.05] and incidence of treatment-related complications (grade 1-2, 30.0% vs 35.7%, P>0.05) between the two groups, respectively. Patients in the two groups had none of serious operation-related complications, and only some patients had mild clinical manifestations, such as postoperative pain and bleeding at the puncture point. Conclusions:For patients with longer duration of hemodialysis and superior vena cava stenosis and occlusion treated with angioplasty, the clinical effect of TCC within one year is equivalent to that of hemodialysis patients without angioplasty.

Distribution of pathogen species and antibiotic resistance of pathogens from intravenous catheter-related bloodstream infections in pediatric intensive care unit / 中华实用儿科临床杂志

Objective To investigate the clinical characteristics,distribution and drug sensitivity of pathogens causing intravenous catheter-related bloodstream infections (CRBSIs) in pediatric intensive care unit (PICU) so as to use antibiotics reasonably.Methods All patients with CRBSIs in PICU of Guangdong General Hospital from September 2009 to September 2014 were investigated and the drug resistance profiles of pathogens causing CRBSIs were also analyzed retrospectively.Results Between 2009 and 2014,there were totally 10 834 catheter days and 23 episodes of CRBSIs with an incidence of 2.1 infections per 1 000 catheter days.Catheter indwell time ＜ 7 days in 9 cases (39.1％),8 to 14 days in 10 cases (43.5％),14 to 21 days in 4 cases (17.4％).There were 13 strains (56.6％) of gram-positive bacteria,5 strains (21.7％) of gram-negative bacteria and 5 strains (21.7％) of fungi.The main pathogens causing CRBSIs were coagulase negative Staphylococci (7 strains,30.4％),Staphylococcus aureus (3 strains,13.0％),Candida albicans(3 strains,13.0％),Candida parapsilosis(2 strains,8.7％),and Enterobacter cloacae (2 strains,8.7 ％).The susceptibility to Vancomycin,Linezolid and Teicoplanin of coagulase negative Staphylococ cus such as S.epidermidis and to Imipenem,Piperacillin/Tazobactam,Cefoperazone/ Sulbactam and Amikacin of gram-positive bacteria arrived at 100.0％,respectively.The candida were 100％ susceptible to Amphotericin B,5-Flucytosine,Fluconazole and Voriconazole.Twenty-one cases (91.3％) received antibiotic treatment versus no antibiotic in 2 cases (8.7％).The average number of antibiotic kinds administered on the patients with fungal infection was 4.4,bacteria were 1.4.Ten cases (43.5％) treatment with 1 kind of antibiotic,4 cases (17.4％) with 2,4 cases (17.4％) with 3,5 cases (21.7％) with more than 3.Twenty-two cases (95.7％) cured and 1 case died (4.3％).Conclusions The major species of pathogen causing CRBSIs was coagulase negative staphylococci in PICU.It is critical for clinicians to guard against fungal infection because of prolonged catheter indwelling time and more antibiotics administered before indwelling catheter.It is effective way to prevent the CRBSIs by reasonably using antibiotics and shortening the time of catheter indwelling.Monitoring CRBSIs pathogenic bacteria distribution and drug susceptibility helps reasonable administration of antibiotics in the earlier time.

Effects of Buyang Huanwu decoction on expressions of nuclear factor-κBp65 and its inhibitor in rats with focal cerebral ischemia injury / 中国中西医结合急救杂志

Objective To explore the effects of Buyang Huanwu decoction(BYHWD)on expressions of nuclear factor-κB p65(NF-κBp65)and its inhibitor( I-κB)in signal transduction of NF-κB in brain tissue of rats with focal cerebral ischemia injury. Methods 180 Sprague-Dawley(SD)rats were randomly divided into normal group,sham-operated group,model group,pynolidine dithiocarbamate(PDTC)group,minocycline(MC)group and BYHWD treatment group,each group 30 rats. The rats of PDTC group were given PDTC 100 mg?kg-1?d-1 by intra-peritoneal injection. In MC group,MC was given by filling the stomach,the dose was 2.35 g?kg-1?d-1,the drug solution was prepared by adding the distilled water,and the total volume of drug solution to fill the stomach was kept at the same volume in various groups,thus the concentration of the drug was different. In BYHWD group,BYHWD was given,the dose was reduced to 5 g?kg-1?d-1 according to the body surface area dose conversion formula about people and animals. In sham-operated group and model group,the distilled water was given in the same volume as other drug solution. The protein expression levels of NF-κBp65 and I-κB in ischemic tissues were examined by using immunohistochemical method on the time points 7,14 and 21 days after treatment in each group. Results Compared with model group, the cell numbers with expression of NF-κBp65 in PDTC group,MC group and BYHWD group were significantly decreased along with the prolongation of therapy time,the decrease in number was more and more,until 21 days,it reached the valley level(cell/400 times HP:44.00±6.91,45.33±6.55,18.67±2.14 vs. 126.00±5.78,all P0.05). After treatment for 7 days,the number of cells with positive expression of I-κB protein in BYHWD group was less than that in MC group(cell/400 times HP:55.00±3.40 vs. 72.50±4.29,P0.05),and after treatment for 21 days,the number in BYHWD group was significantly higher than that in MC group(88.83±4.95 vs. 71.17±7.16, P<0.05). Conclusion BYHWD can regulate the expressions of inflammatory cytokine I-κB and NF-κB in signal transduction of NF-κB in ischemic brain tissue to inhibit the inflammatory reaction,thus it has the protective effect on cerebral ischemia.

Effect of Simotang on gastrointestinal motility and expression of vasoactive intestinal peptide in chronical stressed mice / 中华行为医学与脑科学杂志

Objective To explore effect of Simotang on gastrointestinal motility and expression of vasoactive intestinal peptide(VIP) in the hypothalamus, spinal cord and duodenum of chronical stressed mice. Methods Mice were randomly divided into normal, stress and Simotang group( n= 10 in each group), and given a variety of unpredictable chronic mild stress. After 21 days gastric emptying and intestinal propulsion function were measured,the expression of VIP was detected by immunohistochemistry and RT-PCR. Results Compared with mice in normal group( (49.81 ± 8.56)%; (51.02 ± 5.11 )% ), chronic stress increased gastric residual rate( (61.53 ±8.71 ) %; P ＜ 0.05 ) and reduced small intestine propulsion rate ( ( 31.79 ± 2.38 ) %; P ＜ 0. 05 ). There were differences in expressions of VIP positive cells and mRNA in duodenum( (8.8 ± 1.1 )/mm2 and(0. 58 ±0.03) ),hypothalamus ( ( 12.9 ± 1.5 )/mm2 and (0.81 ± 0. 07 ) ) and spinal cord ( ( 12.1 ± 1. 2)/mm2 and (0.76 ± 0.02) )in chronic stress group compared with normal group( (6.5 ± 0. 9)/mm2 and (0.43 ± 0. 04);( 10.8 ± 1.3 )/mm2and (0.57 ± 0.03 ); (9.3 ± 1.5 )/mm2 and (0.53 ± 0. 02 ) respectively). There was not difference in gastric residual rate (52.93 ± 9.15 )%, small intestine rate(48.98 ± 4.38 )% and expressions of VIP positive cells and mRNA in duodenum ( (6.7 ± 0.9)/mm2 and (0.48 ± 0. 05 ) ), hypothalamus ( ( 10. 6 ± 1.4 )/mm2 and ( 0. 61 ± 0. 05 ) )and spinal cord ( (9. 1 ± 1.3)/mm2 and(0.55 ± 0.05 ) ) in Simotang group compared with those in normal group (P ＞ 0.05 ), but there were decreased compared with those in chronic stress group (P ＜ 0.05 ). Conclusion Simotang can regulate expressions of VIP in duodenum, hypothalamusand spinal cord in chronically stressed mice.

Analysis 22 cases of operative treatment of hip dislocation.with aretabular fracture / 中国基层医药

Objective To discuss the early stage treatment of reduction and the curative effect in coxa dislo- cation with acetabular fracture. Methods According to the different type of coxa dislocation with aeetsbular frac- ture,22 patients were given the distinctive operation and drawn reduction. As to 22 cases combination with acetabu- lar fracture and basis of difference type of the fracture were selected the method of suitable operative entrance or in- ner fixation. In this gronp,22 patients were followed up about 6 month to 3 years(average 15.3 months) after opera- tion. According to the standard of America orthopedic surgery study were commented on curative effect. Results Excellent 15 cases,well 4 casea,pass 2 cases,bad 1 case. Total fine rate were 86% .Conclusion Each kinds type of the hip dislocation cases should needed emergency reduction. The operation treatments were selected first method to settlement the bone shatter displacement in acetabular fracture or unsuitable joint and unsteady joint.