Objective To investigate the effect of prunella vulgaris extract on inflammation,macrophage phenotype,and phagocytic ability in septic mice,and analyze whether Toll like receptor 4(TLR4)/nuclear factor-κB(NF-κB)signaling pathway involved in its mechanism.Methods C57BL/6 mice were divided into the control group,the model group and the prunella vulgaris extract low(25 mg/kg),medium(50 mg/kg)and high(100 mg/kg)dose groups.Except for the control group,all other groups of mice were injected intraperitoneally with lipopolysaccharide(LPS)to prepare sepsis model.Each group was given corresponding medication by gavage.After 24 hours of administration,serum tumor necrosis factor-α(TNF-α),interleukin(IL)-1β,high mobility group protein B1(HMGB1),IL-10 levels,the proportion of M1 type(CD11b+F4/80+)and M2 type(CD206+F4/80+)macrophages in peritoneal macrophages,the phagocytotic capacity of macrophages,the expression of inducible nitric oxide synthase(iNOS)messenger RNA(mRNA)and arginase 1(Arg1)mRNA in peritoneal macrophages and expression levels of TLR4,NF-κB p65 and their phosphorylated proteins in macrophages were detected.Results Compared with the control group,serum TNF-α,IL-1β,HMGB1,proportion of M1 type macrophages in abdominal cavity,mean fluorescence intensity and phagocytotic capacity of macrophages,iNOS mRNA,TLR4,phosphorylated NF-κB p65(p-NF-κB p65)/NF-κB p65 protein expression were increased in the model group(P＜0.05).IL-10,proportion of M2 type macrophages and Arg1 mRNA expression were decreased(P＜0.05).Compared with the model group,serum TNF-α,IL-1β,HMGB1,proportion of M1 type macrophages in abdominal cavity,iNOS mRNA,TLR4,p-NF-κB p65/NF-κB p65 protein expression were decreased successively in the prunella vulgaris extract low,medium and high dose groups(P＜0.05).IL-10,proportion of M2 macrophages,mean fluorescence intensity and phagocytotic capacity of macrophages and Arg1 mRNA expression were increased successively(P＜0.05).Conclusion By inhibiting TLR4/NF-κB pathway,prunella vulgaris extract may inhibit the polarization of peritoneal macrophages into M1 type and promote their polarization to M2 type,enhance macrophage phagocytic ability and alleviate LPS induced inflammatory response in septic mice.

Objective:To observe the effect of immunofluorescence double staining for foamy macrophages and Mycobacterium tuberculosis (MTB) in paraffin-embedded tissue of clinical tuberculous wound, in comparison with three routine staining methods. Methods:The experimental method was used. From April 2019 to May 2020, 10 patients with tuberculous wound (5 males and 5 females, aged 28-77 years) meeting the inclusion criteria were treated in the Department of Burns and Plastic & Wound Repair Surgery of Xiang′an Hospital of Xiamen University. The paraffin-embedded wound tissue were collected during extended debridement and preserved in the Department of Pathology of this hospital. Forty paraffin sections were made from the wound tissue of each patient. Hematoxylin-eosin (HE) staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining were performed respectively, with 10 sections in each method. The section rejection rate of four staining methods were calculated. The recognition and detection of wound granuloma tissue in the four staining methods were observed and counted, and the recognition and detection of foamy macrophages in the wound tissue stained with four methods were observed. The MTB detection in the wound granuloma tissue and non-granuloma tissue in the four staining methods were compared. The subtyping and distribution of foamy macrophages and detection rate of MTB in the wound granuloma tissue and non-granuloma tissue, the morphologic clarity of foamy macrophages, as well as the non-specific staining rate and the loss rate of positive reaction of MTB and foamy macrophages by Ziehl-Neelsen and immunohistochemical double staining were compared with those of immunofluorescence double staining. Data were statistically analyzed with Fisher′s exact probability test, one-way analysis of variance, independent sample t test and Wilcoxon signed rank test. Results:The section rejection rate of HE staining, immunohistochemical staining, Ziehl-Neelsen and immunohistochemical double staining, and immunofluorescence double staining were 3% (3/100), 1% (1/100), 6% (6/100), and 2% (2/100), respectively. There was no statistically significant difference among the four groups ( P=0.26). All the four staining methods could identify granuloma tissue, and the number of granuloma structures was similar ( F=1.284, P=0.28). All the four staining methods were able to identify foamy macrophages in the wound tissue, which was detected in each section. No MTB was observed in the wound granuloma tissue or non-granuloma tissue by HE staining or immunohistochemical staining. MTB was observed distributing in the wound granuloma tissue and non-granuloma tissue by Ziehl-Neelsen and immunohistochemical double staining and immunofluorescence double staining, and most MTB distributed in the wound granuloma tissue. Ziehl-Neelsen and immunohistochemical double staining could not distinguish foamy macrophages engulfed MTB from that non-engulfed MTB. Immunofluorescence double staining showed that foamy macrophages engulfed MTB mostly distributed in the wound granuloma tissue, and the foamy macrophages non-engulfed MTB mostly distributed in the wound non-granuloma tissue. The detection rates of MTB in wound granuloma and non-granuloma tissue in immunofluorescence double staining were (89.00±0.08)% and (82.67±0.05)%, respectively, which were significantly higher than (54.56±0.14)% and (44.44±0.13)% in Ziehl-Neelsen and immunohistochemical double staining ( t=-12.495, -7.961, P<0.01). Compared with that of Ziehl-Neelsen and immunohistochemical double staining, immunofluorescence double staining showed better foamy macrophages clarity in wound tissue ( Z=-3.162, P<0.01). The nonspecific staining rate and positive reaction loss rate of MTB and foamy macrophages in wound tissue of immunofluorescence double staining were (9.11±0.07)% and (9.22±0.07)%, respectively, which were significantly lower than (20.67±0.06)% and (44.00±0.12)% of Ziehl-Neelsen and immunohistochemical double staining ( t=4.569, 15.519, P<0.01). Conclusions:Compared with HE staining, immunohistochemical staining, and Ziehl-Neelsen and immunohistochemical double staining, the immunofluorescence double staining is easy to operate, giving clear and intuitive images. It allows accurate imaging co-localization of MTB and foamy macrophages in paraffin-embedded tissue of clinical tuberculous wound.

OBJECTIVETo investigate the relationship of serum metalloproteinase with the severity of liver fibrosis and inflammation.

METHODSA total of 88 patients with HBV-related liver fibrosis and early cirrhosis were enrolled from six hospitals. Serum fibrosis markers including hyaluronic acid (HA), IV collagen (IV-C), aminoterminal propeptide of type III procollagen (PIIIP), laminin (LN), matrix metalloproteinases (MMP) 1, 2, 9 and tissue inhibitors of metalloproteinase (TIMP) 1, 2 levels were determined. Liver biopsies were assessed according to a modified Scheuer and Chevallier's scoring system.

RESULTSSerum TIMP1 (r=0.540) and MMP2 (r=0.314) were correlated positively with the degree of hepatic fibrosis, whereas serum MMP1 (r=-0.495) was correlated negatively. By receiver operating curve analysis (ROC), the sensitivity to distinguish the fibrosis stage 2 from stage 1 was 90.5% and the specificity was 52.0% if the cut-off value of MMP1 was 13.96 ng/ml, and the sensitivity was 91.6% and the specificity was 64.0% if the cut-off value of TIMP1 was 76.84 ng/ml. The sensitivity to distinguish cirrhosis (stage 4) from fibrosis (stage 3) was 70.7% and specificity was 80.9% if the cut-off value of MMP1 was 6.86 ng/ml, and the sensitivity was 60.5% and the specificity was 92.3% if the cut-off value of TIMP1 was 210.04 ng/ml.

CONCLUSIONSerum TIMP1, MMP1, MMP2 levels and TIMP1/MMP1 ratio could be used as serum fibrosis markers.

Adult ; Biomarkers ; blood ; Female ; Hepatitis B, Chronic ; blood ; complications ; Humans ; Liver Cirrhosis ; blood ; virology ; Male ; Matrix Metalloproteinase 1 ; blood ; Matrix Metalloproteinase 2 ; blood ; Middle Aged ; Tissue Inhibitor of Metalloproteinase-1 ; blood

OBJECTIVESTo further assess the clinical antifibrotic efficacy of Cpd 861 on chronic hepatitis B related fibrosis and early cirrhosis using a randomized, double blind, and placebo controlled clinical trial.

METHODSTotal 136 patients with HBV-related fibrosis and early cirrhosis were allocated randomly into Cpd 861 treatment group and placebo group for 24 weeks treatment. Serum fibrosis markers including hyaluronic acid (HA), IV collagen (IV-C), amino terminal propeptide of type III procollagen (PIIIP), and laminin (LN) and serum MMP1, 2, 9, TIMP1, 2 level were determined before and after 24 weeks treatment. Liver biopsies before and after 24 weeks of treatment were assessed according to modified Scheuer and Chevallier's scoring system.

RESULTSTotal 52 patients in Cpd 861 treatment group and 50 patients in placebo-controlled group completed the 6 months. ALT level decreased from 68.2 U/L+/-68.6 U/L to 45.9 U/L+/-26.1 U/L, AST level decreased from 60.4 U/L+/-62.6 U/L to 46.7 U/L+/-39.0 U/L (P < 0.05) after 24 weeks treatment, whereas there was no significant change in placebo group (ALT: 65.3 U/L+/-48.3 U/L to 85.4 U/L+/-115.5 U/L; AST: 60.4 U/L+/-44.6 U/L to 77.6 U/L+/-89.6 U/L, P > 0.05). Serum fibrosis markers, including HA, IV-C, PIIIP, and LN were decreased after treatment, but there is no statistically significant compared with placebo group. Compared with placebo group, serum TIMP1 and MMP9 level decreased significantly (TIMP1 172.0 ng/ml+/-79.6 ng/ml vs 133.5 ng/ml+/-66.8 ng/ml; MMP9 116.1 ng/ml+/-88.2 ng/ml vs 80.4 ng/ml+/-79.0 ng/ml), and the ratio of TIMP1/MMP1 (48.3+/-96.3 vs 19.9+/-28.0) were also decreased after 861 treatment. In patients treated with Cpd 861, hepatic inflammatory score (from 14.0+/-6.0 to 10.2+/-6.1), fibrosis score (from 11.9+/-6.5 to 8.2+/-4.5), and relative content of collagen (from 18.9%+/-9.5% to 14.9%+/-8.4%) decreased significantly. In contrast, there was no significant change in placebo group. The reversal (fibrosis score decrease > or = 2) rate of fibrosis in Cpd 861 group was 38.9% in S2, 53.3% in S3 (precirrhotic) and 78.6% in S4 (cirrhosis), significantly higher than those in placebo group (14.3%, 25.0%, and 41.7%, respectively). The overall reversal rate was 52.0% in Cpd 861 group, and 20.0% in placebo group (P < 0.05). No serious adverse effects were observed during Cpd 861 treatment.

CONCLUSIONLiver fibrosis and early cirrhosis due to HBV infection in man could be definitely reversed by herbal remedy Cpd 861.

Adolescent ; Adult ; Aged ; Collagen Type IV ; blood ; Double-Blind Method ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Hepatitis B, Chronic ; complications ; drug therapy ; Humans ; Hyaluronic Acid ; blood ; Liver ; pathology ; Liver Cirrhosis ; blood ; drug therapy ; etiology ; Liver Function Tests ; Male ; Middle Aged ; Phytotherapy

OBJECTIVETo investigate the bonding properties and interface characterization of a domestic 3mol yttrium-stabilized tetragonal zirconium polycrystal (3Y-TZP) framework fired on with 4 different veneering ceramics for zirconia.

METHODS4 different commercial veneering ceramics for zirconia (VITA VM9, SHOFU VINTAGE ZR, IPS e.max Ceram, Cercon ceram kiss) were sintered on 3Y-TZP rectangulars (15 mmx5 mmx5 mm) according to the manufacturers' instructions for shear bond strength test, a metal-ceramic system(Ni-Cr alloy/VITA VMK95) was fabricated in the same type as a control group. Two bilayered specimens (3Y-TZP/VITA VM9, Ni-Cr/VMK95) were prepared for scanning electron microscope (SEM) and energy distribution spectrum (EDS).

RESULTSThe values of shear bond strength test were (18.83 +/- 1.77) MPa for 3Y-TZP/VITA VM9, (23.83 +/- 7.05) MPa for 3Y-TZP/SHOFU VINTAGE ZR, (17.87 +/- 2.30) MPa for 3Y-TZP/IPS e.max Ceram, (22.26 +/- 7.45) MPa for 3Y-TZP/Cercon ceram kiss, (20.55 +/- 5.13) MPa for Ni-Cr alloy/VITA VMK95. There was no statistically significant between all-ceramic groups and the control group (P > 0.05). The failure modes in all-ceramic groups showed predominately adhesive at the interface. SEM showed the 3Y-TZP/VITA VM9 contacted tightly at the interface, while EDS detected Si element diffused into 3Y-TZP material.

CONCLUSIONThe results indicate that domestic 3Y-TZP has a good interface compatibility with 4 commercial veneering ceramics, as a dental framework material, it can satisfy the clinical requirements.

Ceramics ; Dental Bonding ; Dental Porcelain ; Dental Stress Analysis ; Dental Veneers ; Materials Testing ; Yttrium ; Zirconium

Animals ; Humans ; Leptin ; genetics ; physiology ; Liver Cirrhosis ; etiology ; RNA, Messenger ; analysis ; Receptors, Cell Surface ; analysis ; physiology ; Receptors, Leptin ; Transforming Growth Factor beta ; genetics ; Transforming Growth Factor beta1

OBJECTIVETo study the targeted point and mechanism of the function of the blood-activating and stasis-removing Chinese drugs, Paeoniae Radix 801(PR801) in its cardiovascular protective effects and its specific binding with endothelin 1 (ET-1) as well as the dynamics of the two's interactive function by means of using affinity biosensors: IAsys Plus and quartz crystal microbalance (IAQCM).

METHODSET-1 was immobilized on the surfaces of IAQCM by using the new surface modification methods. The PR801 in the solution was detected by modified substrates and the specific binding between PR801 and ET-1 was studied.

RESULTSThe curves went up or down after adding PR801. There is specific binding between PR801 and ET-1. The bound mass were 0.458 ng/mm(2) and 133.54 ng/cm(2), respectively. There exists relatively good stability with these two methods.

CONCLUSIONThe affinity biosensors: IAQCM can be used to study the interaction mechanism between PR801 and ET-1, providing a new way to study the interaction mechanism of TCM. PR801 can bind ET-1 specifically in the experiments. Therefore, ET-1 is another target that PR801 can bind specifically besides thromboxane A(2).

Biosensing Techniques ; standards ; Blood Circulation ; drug effects ; Drugs, Chinese Herbal ; metabolism ; therapeutic use ; Endothelin-1 ; metabolism ; Humans ; Quartz

OBJECTIVETo investigate the effect of ketogenic diet (KD) on the clinical and electroencephalogram features in children with pharmacoresistant epileptic encephalopathy.

METHODThirty-one children (19 boys, 12 girls) aged 7 months to 7 years (mean 2 years 5 month) with epilepsy refractory to conventional antiepileptic drugs (AEDs) were included in this study. In addition to their original AED treatment, the children were assigned to different ketogenic diets based on their age. The prospective electro-clinical assessment was performed prior to the KD and then one week, one month and again 3 months after the initiation of therapy, respectively.

RESULTThe reduction of seizure frequency in 52%, 68% and 71% of all patients exceeded 50% one week, one month and three months after KD treatment respectively. KD is particularly effective in myoclonic astatic epilepsy (MAE; Doose Syndrome) and West syndrome with 100% and 81.25% of the patients having a greater than 50% seizure reduction, respectively. After 3 months of KD treatment, more than 2/3 patients experienced a reduction in interictal epileptiform discharges (IEDs) and improvement in EEG background.

CONCLUSIONThe clinical and electroencephalographic improvement confirms that KD is beneficial in children with refractory epilepsy.

Anticonvulsants ; therapeutic use ; Brain ; diagnostic imaging ; physiopathology ; Child ; Child, Preschool ; Diet, Ketogenic ; methods ; Dietary Fats ; administration & dosage ; Electroencephalography ; Epilepsy ; diagnosis ; diet therapy ; drug therapy ; Female ; Humans ; Infant ; Intellectual Disability ; diet therapy ; drug therapy ; Lennox Gastaut Syndrome ; Male ; Radiography ; Retrospective Studies ; Spasms, Infantile ; diet therapy ; drug therapy ; Syndrome ; Time Factors ; Treatment Outcome

OBJECTIVETo study the effect of astragalus injection on U937 leukemia cells proliferation and apoptosis and relevant molecular mechanisms.

METHODSLeukemia cell line U937 cells were treated with different concentrations of astragalus (62.5, 125, 250, 500, 1 000 μg/mL). The U937 cells without astragalus treatment were used as the control group. The ability of cell proliferation was measured by MTT method. Flow cytometry was used to explore cell apoptosis. The cell morphology changes were observed under a fluorescent microscope by dyeing Hoechst33258. mRNA expression of c-myc and p27 in U937 cells which was exposed in 1 000 μg/mL astragalus after 0, 12, 24 and 48 hours was detected by reverse transcription polymerase chain reaction (RT-PCR).

RESULTSVarious concentrations of astragalus injection inhibited U937 cell proliferation effectively compared with the control group (P<0.05). They also induced U937 cells apoptosis and the apoptosis rate reached to (63 ± 4)% in the 1 000 μg/mL astragalus treatment group. mRNA expression level of c-myc was gradually declined and p27 mRNA expression was gradually increased with astragalus treatment time (P<0.01).

CONCLUSIONSAstragalus injection may inhibit proliferation and induce apoptosis of leukemia cell line U937 in vitro. This contributes to down-regulation of c-myc expression and up-regulation of p27 expression.

Apoptosis ; drug effects ; Astragalus Plant ; Cell Proliferation ; drug effects ; Cyclin-Dependent Kinase Inhibitor p27 ; genetics ; Genes, myc ; Humans ; Injections ; U937 Cells

OBJECTIVETo evaluate the efficacy and safety of pegylated interferon alpha 2a (PEG-IFN alpha-2a) in treating patients with chronic hepatitis B.

METHODSeventy-two patients with chronic hepatitis B were assigned to a PEG-IFN alpha-2a (experimental) group (n=42) and an interferon alpha (control) group (n=30) randomly. Each patient in the experimental group received 180 microg PEG-IFN alpha-2a every week. Each patient in the control group received 500 MU interferon alpha every day. All the patients were treated for 48 weeks, and then were followed for another 48 weeks with no treatment.

RESULTSAt the end of the 12th week, the rate of HBeAg negative cases was 30% in the PEG-IFN alpha-2a group, which was much higher than in the control group (x2 = 4.162, P < 0.05). The values of HBeAg and the log value of HBV DNA in the PEG-IFN alpha-2a group were much lower than the values before the treatment (t = 2.689, t = 4.080, P <0.01), but there was no difference between before and after treatment in the control group ( t = 1.229, t = 1.009, P > 0.05). At the end of the 24th week, the rate of HBeAg negative cases in the PEG-IFN alpha-2a group was much higher than that in the control group (x2=6.190, P < 0.05). The value of HBeAg and the log value of HBV DNA in the PEG-IFN alpha-2a group were much lower than in the control group (t=2.215, t=2.122, P < 0.05). At the end of the 48th week, besides the reduction mentioned above, the rate of cases with HBeAg/antiHBe seroconversion and normalization of ALT and complete responsiveness in the PEG-IFN alpha-2a group were all much higher than those in the control group (x2=5.771, x2=5.617, x2=5.308, P < 0.05). At the end of 48 weeks with no treatment, all the parameters mentioned above in the PEG-IFN alpha-2a group were much better than those in the control group and they remained so, but they were different in the control group (x2=11.943, t=3.439, t=6.111, x2=9.930, x2=9.522, x2=7.920, P < 0.01). Nine patients in the PEG-IFN alpha-2a group had liver biopsies before their treatment and also at the end of their treatment. The expressions of HBsAg and HBcAg were decreased at the end of the treatment. The rate of expression of HBsAg in the liver tissues before the treatment was 88.9% but only 22.2% at the end of the treatment (x2=8.001, P < 0.01). The rate of expression of HBcAg in the livers before treatment was 66.7% but only 33.3% at the end of the treatment. Before and at the end of the PEG-IFN alpha-2a treatment, there were no significant changes in the degrees of inflammation and fibrosis and the quantity of collagen in the liver tissues. Three patients in the PEG-IFN alpha-2a group (10%) were HbsAg negative. Two of them were found so at the end of 32 weeks with treatment and one patient was found at the end of 24 weeks with no treatment, but there were no HBsAg negative patients in the control group. The adverse reactions that occurred in the PEG-IFN alpha-2a and in the control groups were similar.

CONCLUSIONPEG-IFN alpha-2a was effective in inhibiting HBV replication. The effect of PEG-IFN alpha-2a was lasting. PEG-IFN alpha-2a was well tolerated during our treatment.

Adolescent ; Adult ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B, Chronic ; drug therapy ; Humans ; Interferon-alpha ; therapeutic use ; Male ; Middle Aged ; Polyethylene Glycols ; therapeutic use ; Recombinant Proteins ; Young Adult