This paper clarified the scientific connotation of the changes in cold and heat properties of Arisaematis Rhizoma and Arisaema Cum Bile through investigating the changes of substance and energy metabolism after drug intervention in the rats with normal and cold/heat syndrome, so as to improve the method of evaluating the drug properties of Chinese medicine. After one week of adaptive feeding, healthy male SD rats were randomly divided into three parts: normal rats, heat syndrome rat models, and cold syndrome rat models. Through ice water bath and oral euthyrox(120 μg·kg~(-1)), the models of cold syndrome and heat syndrome were induced, respectively. The models were made at 9:00 am. and administrated by gavage at 3:00 pm. every day. All administration groups were administrated with Arisaematis Rhizoma and Arisaema Cum Bile decoction, respectively, and the blank group was given the same dose of normal saline. After continuous administration for 15 d, the rats were anesthetized by chloral hydrate, blood was taken from abdominal aorta, and the hearts and livers were removed and stored at-80 ℃. The changes in the body weight and anal temperature of rats during administration were detected, and the liver coefficient of rats was detected after removing the liver. Enzyme-linked immunosorbent assay(ELISA) was adopted to detect the expression level of the indexes related to substance and energy metabolism in liver and heart of rat, and Western blot was used to detect the expression of key proteins in AMPK/mTOR signaling pathway for further verification. The results showed that Arisaematis Rhizoma enhanced the expression level of enzymes related to substance and energy metabolism in the normal and cold and heat syndrome rat models, and increased anal temperature, which exhibited warm(hot) drug property. Arisaema Cum Bile inhibited the level of substance and energy metabolism in rats, and reduced anal temperature, which showed cold(cool) drug property. Chinese Pharmacopoeia has recorded "Arisaematis Rhizoma has warm property and Arisaema Cum Bile has cool property", which is consistent with the phenomenon in this study. Therefore, it is feasible to evaluate the drug properties of Chinese medicine based on the substance and energy metabolism of normal and cold/heat syndrome model rats, which completes the method of evaluating drug properties of Chinese medicine.
AMP-Activated Protein Kinases ; Animals ; Arisaema/chemistry* ; Bile ; Chloral Hydrate ; Cold-Shock Response/drug effects* ; Drugs, Chinese Herbal/therapeutic use* ; Energy Metabolism ; Heat Stroke/therapy* ; Hot Temperature ; Male ; Rats ; Rats, Sprague-Dawley ; Saline Solution ; Syndrome ; TOR Serine-Threonine Kinases ; Thyroxine ; Water

OBJECTIVETo investigate the protective effect of prostaglandin E1 (PGE-1) against brain injury induced by hyperoxia in neonatal rats and observe the changes in the expression of glucose-regulated protein 78 (GRP78) and cyclophosphamide, doxorubicin, vincristine, and prednisone (CHOP), and to provide a theoretical basis for the clinical application of PGE-1 in the treatment of neonatal brain injury induced by hyperoxia.

METHODSSixty neonatal Wistar rats were randomly divided into air control group, hyperoxic brain injury model group, and hyperoxic brain injury+PGE-1 group. All rats except those in the air control group were treated to establish a hyperoxic brain injury model. From the first day of modeling, the rats in the hyperoxia brain injury+PGE-1 group were intraperitoneally injected with PGE-1 2 μg/kg daily for 7 consecutive days, while the other two groups were treated with normal saline instead. The water content of brain tissue was measured; the pathological changes of brain tissue were evaluated by hematoxylin-eosin staining; the apoptosis of brain cells was assessed by nuclear staining combined with TUNEL staining; the protein expression of GRP78 and CHOP in brain tissue was measured by Western blot.

RESULTSThe water content of brain tissue in the hyperoxic brain injury model group was significantly higher than that in the hyperoxic brain injury+PGE-1 group and air control group (P<0.05); the water content of brain tissue in the hyperoxic brain injury+PGE-1 group was significantly higher than that in the air control group (P<0.05). The pathological section of brain tissue showed inflammatory cell infiltration and mild cerebrovascular edema in the brain parenchyma in the hyperoxic brain injury model group; the periparenchymal inflammation and edema in the hyperoxic brain injury+PGE-1 group were milder than those in the hyperoxic brain injury model group. The apoptosis index of brain tissue in the hyperoxic brain injury model group was significantly higher than that in the hyperoxic brain injury+PGE-1 group and air control group (P<0.05); the apoptosis index of brain tissue in the hyperoxic brain injury+PGE-1 group was significantly higher than that in the air control group (P<0.05). The protein expression of GRP78 and CHOP in brain tissue was significantly higher in the hyperoxic brain injury model group than in the hyperoxic brain injury+PGE-1 group and air control group (P<0.05); the protein expression of GRP78 and CHOP was significantly higher in the hyperoxic brain injury+PGE-1 group than in the air control group (P<0.05).

CONCLUSIONSPGE-1 has a protective effect against hyperoxia-induced brain injury in neonatal rats, which may be related to the inhibition of cell apoptosis by down-regulating the expression of GRP78 and CHOP.

Alprostadil ; therapeutic use ; Animals ; Animals, Newborn ; Apoptosis ; drug effects ; Brain ; pathology ; Brain Injuries ; metabolism ; pathology ; prevention & control ; Heat-Shock Proteins ; analysis ; Hyperoxia ; complications ; Neuroprotective Agents ; therapeutic use ; Rats ; Rats, Wistar ; Transcription Factor CHOP ; analysis

OBJECTIVETo observe the effect of Cornus Officinalis total glycosides (COTG) and Cornus polysaccharides (CP) on myocardial mitochondria and expression levels of glycogen synthase kinase-3β (GSK-3β) of acute myocardial infarction (AMI) rats.

METHODSThe AMI rat model was established by ligating the left anterior descending branch of coronary artery. Rats were divided into 5 groups according to random digit table, i.e., the sham-operation group, the model group, the COTG prevention group, the CP treatment group, the COTG treatment group, 12 in each group. Normal saline was administered to rats in the normal control group and the model group by gastrogavage. Corresponding medication was respectively administered to rats in the rest 3 groups by gastrogavage. The cardiac function was detected by echocardiography and hemodynamics. The infarct size was determined by Masson trichrome staining. The expression of mitochondrial biogenesis genes such as a subunit of peroxisome proliferators-activated receptor-γ coactivator-1 (PGC-1α), PGC-1β, nuclear respiratory factor-1 (NRF-1), and GSK-3P mRNA were detected by Real-time PCR.

RESULTSCompared with the sham-operation group, the myocardial infarction size increased, cardiac function decreased, the expression of PGC-1α, PGC-1β, and NRF-1 mRNA decreased, and the expression of GSK-3β mRNA increased (all P <0. 05). Compared with the model group, myocardial infarction sizes were reduced, cardiac function was improved, the expression of NRF-1 mRNA was elevated in the COTG prevention group, the CP treatment group, the COTG treatment group; the expression of the PGC-1α and PGC-1β mRNA was elevated in the COTG prevention group and the CP treatment group; the expression of GSK-3β mRNA was reduced in the CP treatment group (all P <0. 05). Compared with the CP prevention group, fractional shortening (FS) and aortic systolic blood pressure (SBP) increased in the CP treatment group; ejection fraction (EF) decreased in the CP treatment group; the expression of PGC-1α, PGC-1β, NRF-1 mRNA were reduced in the the CP treatment group and the COTG treatment group; the expression of GSK-3β mRNA decreased in the CP treatment group (all P <0. 05). Compared with the COTG treatment group, FS, EF, left ventricular end systolic pressure (LVESP), SBP, and the expression of GSK-3β mRNA were reduced in the CP treatment group (P <0. 05).

CONCLUSIONSCOTG and CP could improve cardiac function, reduce the myocardial infarction area, and promote biogenesis of myocardial mitochondria. Their protective effects on the mitochondria of cadiocytes might be achieved by GSK-3β signalina pathway.

Animals ; Cornus ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Glycogen Synthase Kinase 3 ; Glycogen Synthase Kinase 3 beta ; Glycosides ; Heat-Shock Proteins ; Mitochondria, Heart ; physiology ; Myocardial Infarction ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Polysaccharides ; Protective Agents ; pharmacology ; therapeutic use ; RNA, Messenger ; Rats ; Rats, Sprague-Dawley ; Transcription Factors

OBJECTIVETo observe the effect of Chinese drugs for strengthening Pi, harmonizing Wei, and dispersing blood stasis (CDSPHWDBS) on the expression of gastric mucosal heat shock protein 70 (HSP70) in chronic atrophic gastritis (CAG) patients.

METHODSA total of 100 CAG patients were assigned to the control group and the treatment group by random digit table, 50 in each group. Patients in the control group took Folic Acid Tablet 10 mg each time, 3 times per day. Those in the treatment group took CDSPHWDBS, 100 mL each time, once per day. The treatment course was 6 months for all. Clinical symptoms and signs, endoscopic and histopathological changes were observed before and after treatment in the two groups. The expression of gastric mucosal HSP70 in CAG patients was determined using SP immunohistochemistry. Data were collected by HPIAS-1 000 pathological graphic analysis system, and its expression semi-quantitatively analyzed.

RESULTSThe total effective rate of clinical Chinese medical symptoms and signs was 88. 0% (44/50 cases) in the treatment group and 56. 0% (28/50 cases) in the control group, with significant difference between the two groups (P <0. 01). The improvement rate of endoscopic manifestations such as congestion and edema, erosion, bile regurgitation, pale gastric mucosa, exposed blood vessels, particles proliferation in the treatment group were superior to those in the control group (P <0. 05). The total effective rate of atrophy was 80. 0% (40/50 cases) in the treatment group and 54. 0% (27/50 cases) in the control group, with significant difference between the two groups (P<0. 01). The effective rate of intestinal metaplasia was 75. 0% (12/16 cases) in the treatment group and 33.3% (5/15 cases) in the control group, with significant difference between the two groups (P < 0. 05). The optical density value of gastric mucosal HSP70 was significantly elevated in the two groups after treatment (both P <0. 05). It was higher in the treatment group than in the control group after treatment with significant difference (P <0. 01).

CONCLUSIONCDSPHWDBS had obvious effect in treatment of CAG and could improve pathological changes of precancerous lesions possibly by promoting the expression of gastric mucosal HSP70 in CAG patients.

Drugs, Chinese Herbal ; therapeutic use ; Gastric Mucosa ; metabolism ; Gastritis, Atrophic ; drug therapy ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Immunohistochemistry ; Medicine, East Asian Traditional

OBJECTIVETo investigate the protective effect of ulinastatin (UTI) against acute lung injury induced by heatstroke in mice.

METHODSSixty C57/BL6 mice were randomly divided into 6 groups, with 10 mice in each: control group, heatstroke group, UTI pretreatment group, saline pretreatment group, UTI post-treatment group, saline post-treatment group. The control mice were housed at a controlled room temperature of (22∓1) degrees; celsius, and the other groups were placed inside a temperature and humidity controlled chamber pre-set at 37 degrees; celsius and 60%. The two UTI groups were intraperitoneally injected with UTI at 5×10(4) U/kg 10 min before or after heat stress, and the two saline groups were given then equal amounts of saline in the same manner. The core body temperature of mice was monitored by a mercury thermometer every 30 min in the first 1.5 h during heating. The core temperature was measured, then every 15 min until it reached 42.7 degrees; celsius, which was taken as the onset of heatstroke. The animals were allowed to recover passively at ambient temperature for 6 h. The lung histopathological changes, protein concentration in BALF, lung wet/dry weight ratios, lung water content, and pulmonary microvascular permeability were assayed after 6 h of recovery at 37 degrees;celsius.

RESULTSCompared with the control group, the heatstroke model group and two saline groups displayed more severe lung damage and pathological morphology changes, and the lung wet/dry weight ratio, protein concentration in BALF, lung water content and pulmonary microvascular permeability were also significantly increased. These effects were significantly alleviated in UTI treated group. Pretreat ment with UTI significantly prolonged the time to Tc≥42.7 degrees; celsius but had no effect on lung injury induced by heatstroke.

CONCLUSIONUTI can reduce the pulmonary edema and inflammatory exudation in acute lung injury caused by heatstroke.

Acute Lung Injury ; drug therapy ; physiopathology ; Animals ; Body Temperature ; Bronchoalveolar Lavage Fluid ; chemistry ; Edema ; prevention & control ; Glycoproteins ; therapeutic use ; Heat Stroke ; physiopathology ; Lung ; pathology ; Mice ; Mice, Inbred C57BL

BACKGROUNDTo develop a vaccine-based immunotherapy for sarcoma, we evaluated a mixture of heat shock proteins (mHSPs) as a vaccine for sarcoma treatment in a mouse model. Heat shock protein/peptides (HSP/Ps) are autoimmune factors that can induce both adaptive and innate immune responses; HSP/Ps isolated from tumors can induce antitumor immune activity when used as vaccines.

METHODSIn this study, we evaluated the effects of mHSP/Ps on prophylactic antitumor immunity. We extracted mHSP/Ps, including HSP60, HSP70, GP96, and HSP110, from the mouse sarcoma cell lines S180 and MCA207 using chromatography. The immunity induced by mHSP/Ps was assessed using flow cytometry, ELISPOT, lactate dehydrogenase release, and enzyme-linked immunosorbent assay.

RESULTSOf S180 sarcoma-bearing mice immunized with mHSP/Ps isolated from S180 cells, 41.2% showed tumor regression and long-term survival, with a tumor growth inhibition rate of 82.3% at 30 days. Of MCA207 sarcoma-bearing mice immunized with mHSP/Ps isolated from MCA207 cells, 50% showed tumor regression and long-term survival with a tumor growth inhibition rate of 79.3%. All control mice died within 40 days. The proportions of natural killer cells, CD8+, and interferon-γ-secreting cells and tumor-specific cytotoxic T-lymphocyte activity were increased in the immunized group.

CONCLUSIONSVaccination with a polyvalent mHSP/P cancer vaccine can induce an immunological response and a marked antitumor response to autologous tumors. This mHSP/P vaccine exerted greater antitumor effects than did HSP70, HSP60, or tumor lysates alone.

Animals ; Cancer Vaccines ; therapeutic use ; Female ; Heat-Shock Proteins ; administration & dosage ; Immunotherapy ; methods ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Peptides ; administration & dosage ; Sarcoma, Experimental ; prevention & control ; Vaccination

OBJECTIVETo investigate the protective effect of H2S pretreatment after cerebral schemia/reperfusion injury and its mechanisms in rats.

METHODSThe rat model of global cerebral ischemia/reperfusion injury was established by bilateral common carotid arteries occlusion combined with hemorrhagic hypotension.30 rats were randomly divided into four groups(1)sham group(n=5),in which rats received sham surgery only,with their bilateral vertebral artery and bilateral common carotid artery exposed but without ischemia treatment;(2)global cerebral ischemia/reperfusion model group(IR group,n=5),in which the global cerebral ischemia was induced by 10-min occlusion of bilateral common carotid arteries combined with hypotension;(3)H2S pretreatment group(n=15),in which H2S(12,24,48 Μmol/kg)was intraperitoneally injected before operation;(4)NaCl pretreatment group(n=5),in which the rats were intraperitoneally injected with saline 30 minutes before operation.The activities of superoxide dismutase(SOD)and the levels of malondialdeehyde(MDA)in brain were measured by spectrophotometry.Brain water content was detected.The expression of heat shock protein 70(HSP70) in the hippocampus was determined by Western blotting.

RESULTSThe SOD activities were significant increased in groups pretreated with 12Μmol/kg H2S(P=0.042),24Μmol/kg H2S(P=0.002),and 48Μmol/kg H2S(P=0.000),and the SOD activity was significantly lower in the ischemia group than in the Sham group(P=0.003).The MDA activities in the 24Μmol/kg group(P=0.026)and the 48Μmol/kg group(P=0.015)groups were significantly lower than in the IR group.The brain water content was decreased in H2S pretreatment group(24Μmol/kg and 48 Μmol/kg)compared with IR group(P=0.018,P=0.008),and it was also significantly higher in the IR group than in the sham group(P=0.009).The expression of HSP70 were decreased in H2S pretreatment group(24 Μmol/kg)compared with the IR group(P=0.000),and the expression of HSP70 were significantly higher in the IR group than in HSP70 group(P=0.000).The expression of HSP70 also significantly differed between NaCl group and HSP70 group(P=0.000).

CONCLUSIONH2S has protective effects on cerebral ischemia and reperfusion,which may be achieved by improving SOD activity,removing oxygen free radicals,inhibiting lipid peroxidation,and down-regulating the expression of HSP70 in the hippocampus.

Animals ; Brain Ischemia ; metabolism ; Free Radicals ; metabolism ; HSP70 Heat-Shock Proteins ; metabolism ; Hippocampus ; metabolism ; Hydrogen Sulfide ; administration & dosage ; therapeutic use ; Male ; Random Allocation ; Rats ; Reperfusion Injury ; metabolism ; prevention & control ; Superoxide Dismutase ; metabolism

OBJECTIVETo investigate the clinical value of the expression of glucose regulated protein 78 (GRP78) for assessment of severity, chemoresistance and prognosis in patients with gastric adenocarcinoma ( GC) .

METHODSA cohort of 237 patients with gastric cancer was included in this study. 160 patients of them were treated by D2 radical gastrectomy and adjuvant chemotherapy. The GRP78 expression was detected by immunohistochemistry and 80 patients of them were tested in vitro for cancer chemosensitivity by ATP-tumor chemosensitivity assay (ATP-TCA). In addition, the relationships were analyzed between GRP78 and age, gender, tumor differentiation, invasion, disease stage, lymph node metastasis and chemoresistance as well as disease-free survival (DFS).

RESULTSThe positive rate of GRP78 expression in the gastric adenocarcinoma was 68.8% before the initiation of chemotherapy. The positive GRP78 expression was significantly correlated with tumor invasion depth, poor differentiation, TNM stages, and lymph node metastasis (all P < 0.05), not correlated with gender and age, and high GRP78 expression was associated with the chemoresistance of the gastric cancer cells to chemotherapeutic agents. Negative GRP78 expression was associated with higher sensitivity to both drugs and regimens. The DFS of GRP78-positive group and GRP78-negative group was (53.6 ± 0.9) months and (38.3 ± 0.8) months, respectively (P = 0.041). Interestingly, subgroup analysis revealed that the DFS in GRP78-negative and-positive patients treated with taxane-containing chemotherapy was (58.6 ± 2.6) months and (49.1 ± 2.7) months, respectively, but the difference was statistically not significant (P = 0.111). In contrast, in the subset of GRP78-negative and- positive patients treated with taxane-containing regimens, the DFS was (45.5 ± 1.9) months and (35.1 ± 2.2) months, respectively, showing a significant difference (P = 0.038). In the group of patients with positive GRP78 expression, the patients treated with taxane-containing chemotherapy had a longer DFS [(49.1 ± 2.7) months] than those without that treatment [(35.1 ± 2.2) months], showing a significant difference (P = 0.017). Univariate analysis revealed that DFS was correlated with histological grade, GRP78 expression and lymph node metastasis (all P < 0.05). Multivariate analysis showed that GRP78 expression and TNM staging were independent influencing factors for gastric cancer (both P < 0.05).

CONCLUSIONSThe results of our study suggest that GRP78 may be a novel biomarker for assessment of malignant degree and prediction of chemoresistance in gastric cancer, and may be helpful to chemotherapy planning and prognosis prediction in patients with gastric cancer.

Adenocarcinoma ; drug therapy ; metabolism ; pathology ; surgery ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Biomarkers, Tumor ; metabolism ; Bridged-Ring Compounds ; administration & dosage ; Chemotherapy, Adjuvant ; Disease-Free Survival ; Drug Resistance, Neoplasm ; Female ; Gastrectomy ; Heat-Shock Proteins ; metabolism ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Grading ; Neoplasm Invasiveness ; Neoplasm Staging ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; surgery ; Taxoids ; administration & dosage

This study was aimed to investigate the sensitive index for predicting the sensitivity of patients with multiple myeloma (MM) to glucocorticoid (GC). MM1.S, MM1.R cell lines and 10 MM patients sensitive or non-sensitive to GC were enrolled in this study. The expression of heat shock protein 90 (HSP90) and P23 in peripheral blood mononuclear cells (PBMNC) of MM patients was detected by Western blot, and its relation with GC resistance was explored. The results showed that there were no differences in HSP90, GR and IκB-α expressions among MM1.S, MM1.R, PBMNC of normal persons and MM patients, while there was apparent difference in P23; along with increasing of dexamethasone dose, the survival rate of MM1.S decreased, but that of MM1.R did not change so obviously. Among 7 cases with P23 low expression, combination treatment scheme with GC was effective in 6 cases (86%), uneffective in 1 cases (14%); while among 3 cases with P23 high expression, 1 cases (33%) showed effective to combination treatment with GC, 2 cases (67%) showed uneffective (P < 0.05). It is concluded that the P23 expression may predict the sensitivity of MM patients to GC and may be used as index for guiding clinical treatment.
Aged ; Aged, 80 and over ; Female ; Glucocorticoids ; pharmacology ; therapeutic use ; HSP90 Heat-Shock Proteins ; metabolism ; Humans ; Male ; Middle Aged ; Molecular Chaperones ; metabolism ; Multiple Myeloma ; diagnosis ; drug therapy ; metabolism ; Phosphoproteins ; metabolism ; Tumor Cells, Cultured

Heat-shock protein gp96 associates with antigenic peptides derived from tumor and virus. Exogenous gp96-peptide complexes are taken up by antigen-presenting cells through interaction with its receptor CD91 on the cell surface, and cross-present antigenic peptides to MHC class I molecules by a peptide relay line in the endoplasmic reticulum for specific T-cell activation. Meanwhile, gp96 has been shown to initiate innate immune responses through interaction with toll-like receptor 2 and toll-like receptor 4. Recent studies have shown a gp96-mediated immune balance between CTL and Tregs. With the further understanding of counteracting immunosuppressive mechanisms in gp96-induced cellular immune responses, and establishment of high level production of recombinant gp96 by the yeast, gp96 appears to be a promising candidate for designing effective therapeutic vaccines against tumor and infectious diseases.
Animals ; Communicable Diseases ; therapy ; Heat-Shock Proteins ; immunology ; therapeutic use ; Humans ; Immunotherapy, Active ; methods ; Neoplasms ; therapy ; T-Lymphocytes, Regulatory ; immunology