OBJECTIVETo study the effect of electroacupuncture on nitric oxide synthase (NOS) in rats with cerebral ischemia-reperfusion injury.

METHODSFocal cerebral ischemia-reperfusion model was established using modified intravascular suture technique. The NO content in the brain tissue was detected by nitrite reduction and the expressions of nNOS and iNOS were detected by immunohistochemistry. Eighty rats in this experiment were divided into the normal group, the cerebral ischemia-reperfusion injury model group (as the model group), the cerebral ischemia-reperfusion injury + electroacupuncture group (as the acupuncture group), and the cerebral ischemia-reperfusion injury + phosphatidylinositol 3 kinase (PI3-K) inhibitor group (as the inhibitor group). Each group consisted of twenty rats. Five microL PI3-K inhibitor LY294002 (400 microL) was slowly injected at the lateral cerebral ventricle of rats in the inhibitor group at a constant speed using microinjector according to Konig Klippel atlas of the stereotaxis instrument. Shuigou (DU26) and Chengjiang (RN24) were selected to determine levels of NO and NOS.

RESULTSAfter 24-h ischemia-reperfusion, the NO levels of the hippocampus and the cerebral cortex increased abnormally, and the expressions of nNOS and iNOS increased, showing significant difference when compared with those of the normal group (P<0.05). By electroacupuncture at Shuigou (DU26) and Chengjiang (RN24), the ischemic cerebral ischemia-reperfusion injury neuron loss was inhibited. Meanwhile, the high levels of NO, nNOS and iNOS in the cerebral cortex and the hippocampus were significantly inhibited (P<0.05). The abnormally increased expressions of nNOS and iNOS were reversed, showing significant difference when compared with the model group (P<0.05). But when compared with the normal group, there was no significant difference (P>0.05). The effects of electroacupuncture reversed the abnormally increased NO levels of the hippocampus and the cerebral cortex and expressions of nNOS and iNOS after LY294002 oppressed anti-PI3K to block the TrkA acceptor circuit. The NO levels of the hippocampus and the cerebral cortex and expressions of nNOS and iNOS increased again, showing significant difference when compared with the acupuncture group (P<0.05).

CONCLUSIONSAcupuncture fought against cerebral ischemia and reperfusion in the loss of neurons, at the same time, the abnormal regulation of NOS had reverse effect partly through TrkA/PI3K mediated signal transduction pathway.

Animals ; Brain Ischemia ; metabolism ; Electroacupuncture ; Nitric Oxide ; metabolism ; Nitric Oxide Synthase Type I ; metabolism ; Nitric Oxide Synthase Type II ; metabolism ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; Signal Transduction

Objective To investigate the capability of empathy for pain in migraineurs without aura.Methods Thirty migraineurs without aura and thirty matched healthy controls were recruited.Picturecued Empathy for pain paradigm was used to compare the capability of empathy in the migraine group with that in the control group.Results Compared with the control group,the migraine group had diminished ability to discriminate painful from nonpainful pictures,and the discrimination accuracy was significantly reduced ((2.55±0.61) vs (2.88±0.38);t=-2.505,P=0.01).In the task laterality,there was no difference in discrimination accuracy between two groups(P＞0.05).The rating scores of patients were evidently smaller than those of control group ((3.01±0.52) vs (3.37±0.47);t=-2.827,P=0.006).Pearson correlation analysis showed that the age of migraineurs was negatively correlated with the discrimination accuracy(r=-0.393,P =0.031),and there was no correlation between migraineurs' educational years,disease course,severity,Mini-mental State Examination,Hamilton Anxiety Scale,Hamilton Depression Scale,Verbal Fluency Test,Stroop Test and the idex of empathy for pain (all P＞0.05).Conclusion Migraineurs without aura have deficiency in the capability of empathy for pain.

OBJECTIVETo investigate anticancer effects of 5-fluorouracil (5-FU) combined with CL, extract of Rosa roxburghii Tratt on human endometrial adenocarcinoma cell line (JEC).

METHODSJEC cells cultured in vitro in the logarithmic growth phase were seeded in the culture plate and divided into the control group (RPMI 1640), the positive group (10(-4) mol/L 5-FU), the CL groups (at the dose of 0.01, 0.1, 1, 10, and 100 microg/mL), and the CL (0.01, 0.1, 1, 10, and 100 microg/mL) combined with 5-FU groups. Effects of 5-FU combined with CL on JEC cell growth were drawn and measured by MTT and growth curves. Effects of CL combined with 5-FU on the JEC cell differentiation was analyzed by detecting the reduction capability of nitrobenzene thiocyanate (NBT) and lactate dehydrogenase (LDH) contents in the cultured medium. Effects of CL combined with 5-FU on the JEC cell apoptosis and cell proliferation cycle were detected by acridine orange (AO)/ethidium bromide (EB) fluorescent staining and flow cytometry (FCM).

RESULTSThe proliferation inhibitory effect of CL combined with 5-FU on JEC cells was enhanced when compared with that of CL or 5-FU alone (P<0.05). The percentages of NBT positive JEC cells and apoptotic JEC cells increased in the 5-FU combined with CL groups when compared with 5-FU group or the CL group alone (P<0.05). The LDH concentration of the JEC cell culture supernate decreased in 5-FU combined with CL groups (P<0.05). Furthermore, the percentage of G0-G1 phase JEC cells treated by 5-FU combined with CL was higher than that of 5-FU or CL alone (P<0.05).

CONCLUSIONCL could enhance anticancer effects of 5-FU. Its mechanisms might be correlated with reinforcing the cytotoxicity of 5-FU, inducing cell differentiation and apoptosis, and inhibiting cell proliferation and division.

Adenocarcinoma ; pathology ; Cell Differentiation ; drug effects ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Endometrial Neoplasms ; pathology ; Female ; Fluorouracil ; pharmacology ; Herb-Drug Interactions ; Humans ; Plant Extracts ; pharmacology ; Rosa ; chemistry

Child ; Histiocytosis, Sinus ; diagnosis ; drug therapy ; Humans ; Lymphatic Diseases ; diagnosis ; drug therapy ; Male

The chemical investigation on Ganoderma philippii led to the isolation of sixteen compounds by silica gel and Sephadex LH-20 column chromatography. On the basis of spectroscopic data analyses, their structures were elucidated as 2, 5-dihydroxyacetophenone (1), methyl gentisate (2), (S) -dimethyl malate (3), muurola-4, 10 (14) -dien-11beta-ol (4), dihydroepicubenol (5), 5-hydroxymethylfuran carboxaldehyde (6), ergosta-7, 22E-dien-3beta-ol (7), ergosta-7, 22E-dien-3-one (8), ergosta-7, 22E-diene-2beta, 3alpha, 9alpha-triol (9), 6/beta-methoxyergo-sta-7, 22E-dien-3beta, 5alpha-diol (10), ergosta-4, 6, 8(14), 22E-tetraen-3-one (11), ergosta4, 6, 8-(14), 22E-etetraen-3beta-ol (12), 5alpha, 8alpha-epidioxy-ergosta-6, 22E-dien-3beta-ol (13), 7alpha-methoxy-5alpha, 6alpha-epoxyergosta-8-(14), 22E-dien-3beta-ol (14), ergosta-8, 22E-diene-3beta, 5alpha, 6beta, 7alpha-tetraol (15), and ergosta-5, 23-dien-3beta-ol, acetate (16). All the compounds were obtained from this fungus for the first time, and compounds 4 and 5 were isolated from the Ganoderma genus for the first time.
Ganoderma ; chemistry ; Medicine, Chinese Traditional ; Organic Chemicals ; analysis ; isolation & purification

Objective The invasion and metastasis of colon cancer often leads to treatment failure and mortality in patients . Our research is to investigate the influence of FoxM 1 to malignant human colon cancer line . Methods In two human colon cancer lines, the protein and mRNA expression levels of FoxM 1 were analyzed with the application of RT-PCR and Western blot , from which high-expressed HT-29 and low-expressed HCT-116 were determined.The expression of FoxM1 was down-regulated by RNA interfering in HT-29 and up-regulated by constructing overexpression transgenic line in HCT-116.The proliferation of the above cells was assayed by healing method;while the metastasis and invasion ability were examined by Transwell chamber assay . Results Two colon cancer lines were selected with high-expression or low-expression of FoxM1 separately named HT-29 and HCT-116.Application of PEX-2-FoxM1 raised after HCT-116 cells express FoxM1, cell scratches in HCT-116 experimetal group ([70.92 ±1.48]%) compared with HCT-116 control group([16.92 ±4.05]%)and HCT-116 blank control group([16.66 ±2.63]%) will markedly enhance its capabil-ity of healing (P<0.05), Transwell Chambers in membrane cells in HCT-116 experimetal group (186.0 ±6.8) compared with HCT-116 control group(42.0 ±2.0) and HCT-116 blank control grou (37.0 ± 2.2)was increased (P<0.05).On the other hand, the applied pG-PH-shFoxM1 can reduce FoxM1 expression in HT-29 cell, cell scrat-ches healing ability in HT-29 experimetal group ( [ 10 .37 ± 3.86]%) compared with HT-29 control group([34.63 ±2.35]%)and HT-29 blank control group([67.36 ±2.61]%) decreased significantly (P<0.05), Transwell Chambers in membrane cells in HT-29 experimetal group (53.0 ±1.8)compared with HT-29 control group(95.0 ±2.2)and HT-29 blank control grou(118.0 ±4.0) was also reduced (P<0.05). Conclusion The expression of FoxM1 is in close relation to the invasion and metastasis of CRC .The fact that the siRNA interfering FoxM1 could effectively inhibit the proliferation, metastasis and invasion, suggesting FoxM1 could po-tentially be a new molecular target for inhibiting the proliferation of human colon cancer line .

OBJECTIVETo investigate anticancer effects and potential mechanisms of CL, extract of Rosa roxburghii.

METHODIn vitro anticancer effect was observed in Ehrlich's ascites carcinoma (EAC) mice model. Cell toxicity of CL on human endometrial adenocarcinoma cell line JEC (JEC) cells was measured by MTT reduction test and growth curves drawing by trypan blue dye exclusion method. Lactate dehydrogenase (LDH) concentration of cultured medium was detected by auto-biochemistry-meter. Cell differentiation was showed by detection of NBT reduction ability. Apoptosis was showed by AO/EB fluorescent staining and flow cytometer detection. Cell proliferation cycle was detected by flow cytometer.

RESULTComparing with the negative group, life span of EAC mice treated with CL was prolonged (P <0.05), and thymus index and spleen index of them were raised (P <0.05). The inhibitory effect of CL on JEC cells was in concentration-and time-dependent manner. IC50 of CL on JEC cells was 0.05 microg mL(-1) in 96 hours. Growth curves showed right-shift with CL concentration increasing. The number of NBT positive JEC cells increased and the LDH concentration of cultured medium declined with CL increasing. Apoptosis of JEC cells with CL treated was induced in concentration-dependent manner, apoptotic percentage of CL 10 microg mL(-1) on JEC cells was 25.59% in 24 hours. CL arrested JEC cells in G2-M phase (P <0.05).

CONCLUSIONCL has certainly anticancer effects in vivo and in vitro. Anticancer effect of CL in vivo was in relation to enhancing immune function of EAC mice; anticancer mechanisms of CL on JEC cells may be its direct cytotoxic effect, inducing cell apoptosis and inhibiting cell segmentation.

Adenocarcinoma ; metabolism ; pathology ; Animals ; Antineoplastic Agents, Phytogenic ; administration & dosage ; pharmacology ; Apoptosis ; drug effects ; Carcinoma, Ehrlich Tumor ; pathology ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Dose-Response Relationship, Drug ; Drugs, Chinese Herbal ; administration & dosage ; pharmacology ; Endometrial Neoplasms ; metabolism ; pathology ; Female ; Humans ; L-Lactate Dehydrogenase ; metabolism ; Mice ; Plants, Medicinal ; chemistry ; Random Allocation ; Rosa ; chemistry

OBJECTIVETo evaluate the prognostic implication of the killer-immunoglobulin like receptor (KIR) ligand mismatch in HLA mismatched hematopoietic stem cell transplantation (HSCT).

METHODSNinety-four leukemia patients undergoing unmanipulated HLA-mismatched/haploidentical blood and marrow HSCT enrolled this study.

RESULTSMultivariate analysis showed that both KIR ligand mismatch (HR 2.833, CI, 1.286 - 6.241, P = 0.01) and doses of T cells (HR 3.059, CI, 1.292 - 7.246, P = 0.011) were independent risk factors for the acute graft versus host disease (aGVHD). In addition, compared to those without KIR ligand mismatch, patients with KIR ligand mismatch had the more adverse effect of 'high' dose T cells (> 1.48 x 10(5)/kg) on aGVHD (100% vs 63.3%, P = 0.036), and had more incidence of aGVHD with HLA-C mismatch (80.0% vs 57.4%, P = 0.056). Since multivariate analysis demonstrated that high risk leukemia was the only predictor for transplant related mortality (TRM), relapse and overall survival (OS), the effect of KIR ligand mismatch on prognosis in standard and high risk patients was further analyzed. The differences in TRM (50.0% vs 7.6%, P = 0.005) and OS (50.0% vs 88.4%, P = 0.014) between patients with and without KIR ligand mismatch were most striking for standard risk patients.

CONCLUSIONKIR ligand mismatch is a poor prognosis factor for patients underwent HLA mismatched HSCT, and is a useful parameter for donor selection.

Adolescent ; Adult ; Child ; Child, Preschool ; Female ; Graft vs Host Disease ; prevention & control ; HLA Antigens ; immunology ; Hematopoietic Stem Cell Transplantation ; Humans ; Male ; Middle Aged ; Multivariate Analysis ; Prognosis ; Receptors, KIR ; immunology ; Retrospective Studies ; Young Adult

Objective:To analyze the clinical characteristics and causes of death of 80 dead cases with confirmed coronavirus disease 2019 (COVID-19).Methods:The clinical data of 80 dead patients with COVID-19 who were admitted to Renmin Hospital of Wuhan University from January 11 to February 11, 2020 were retrospectively analyzed.The laboratory examination indexes (including white blood cells, lymphocytes, procalcitonin (PCT), lactic acid, D-dimmer, fibrinogen degradation products, N-terminal pro-brain natriuretic peptide (N-proBNP), ultra sensitive-troponin I, lactate dehydrogenase (LDH) and CD4 + T lymphocyte) of the patients at the time of admission were compared with the indexes at the last time before death. Statistical analysis was conducted by using paired t test or Wilcoxon′s signed rank test. Results:The median age was 72 years old of the 80 patients, and 78.75%(63/80) of them were older than 60 years. Thirty-six cases (45.00%) were severe and 44(55.00%) were critical at admission. Fifty-eight cases (72.50%) had underlying diseases. The common underlying diseases were hypertension, diabetes mellitus, coronary atherosclerotic heart disease, and chronic obstructive pulmonary disease. Comparing the patients′ first laboratory tests at admission with those before death, white blood cells increased (8.01(4.86, 12.29)×10 9/L vs 12.55(8.25, 17.66)×10 9/L), lymphocytes decreased (0.70(0.46, 0.88)×10 9/L vs 0.54(0.39, 0.75)×10 9/L), PCT increased (0.20(0.11, 0.74) μg/L vs 1.00(0.20, 1.99) μg/L), lactic acid increased (2.10(1.40, 3.10) mmol/L vs 3.10(2.60, 4.10) mmol/L), D-dimmer increased (4.33(0.97, 18.98) mg/L vs 15.29(5.17, 53.44) mg/L), fibrinogen degradation products increased (15.90(3.58, 76.60) mg/L vs 63.14(21.23, 110.67) mg/L), N-proBNP increased (1 078.00(347.35, 2 996.50) ng/L vs 3 439.50(1 576.00, 9 281.50) ng/L), ultra-sensitive troponin I increased (0.08(0.03, 0.17) μg/L vs 0.33(0.14, 2.47) μg/L), LDH increased (397.00(327.00, 523.50) U/L vs 624.00(481.00, 854.00) U/L) and CD4 + T lymphocyte decreased (137.00(104.00, 168.00)/μL vs 97.00(67.00, 128.00)/μL). The differences between the two groups were all statistically significant ( W=238.00, 1 053.50, 150.00, 152.00, 192.00, 190.00, 108.00, 57.00, 53.00 and 40.00, respectively, all P<0.05). All patients received antiviral and respiratory-support therapy and the main cause of death was respiratory failure caused by intractable hypoxemia and multiple organ failure. Among them, seven cases died in one day hospitalization, and 66 cases died in seven days hospitalization. Conclusions:Elderly patients with a variety of chronic underlying diseases have poor prognosis. It′s essential to pay more attention and deal with the above clinical characteristics at an early stage to improve the outcome of the COVID-19 patients.

OBJECTIVETo study some physicochemical properties of Angelica sinensis polysaccharide-iron complex (APC).

METHODBased on the qualitatively identified reactions of iron (III), the qualitatively identified reactions of APC were found out by comparing hydroxide. The content of iron (III) in APC was determined with iodometry. The stability of APC under physiological pH conditions was judged by titrating APC with sodium hydroxide. The deoxidization of APC was tested with colorimetric analysis.

RESULTAPC showed the qualitatively identified reactions of iron (III). The content of iron (III) in APC ranged from 10% to 40%, and the water-solubility of APC was related to the content of iron (III). The complex was stable at physiological pH from 3 to 12, without precipitation and dissociation. At 37 degrees C, Fe (III) in the complex was completely reduced to Fe (III) by ascorbic acid in about 6 hours.

CONCLUSIONAPC can be qualitatively identified by using the qualitatively identified reactions of iron (III). When its iron (III) content is within 20%-25%, APC has a better ability to dissolve in water. And the complex is stable under physiological pH conditions.

Angelica sinensis ; chemistry ; Chemical Phenomena ; Chemistry, Physical ; Drug Stability ; Ferric Compounds ; administration & dosage ; chemical synthesis ; chemistry ; Hydrogen-Ion Concentration ; Iron ; analysis ; Plants, Medicinal ; chemistry ; Polysaccharides ; administration & dosage ; isolation & purification ; Solubility