Fifty-one male rats (body weights:200—350gm) were raised with laboratory diet, 39 of them were injected once a week with CCl_4 (0.015cc/100gm body weight) in li- quid paraffin through the mesenteric veins and 12 rats without treatment as controls. Rats with 1,2,3,and 4 injections were killed 7 days afterwards.Pieces of liver tissue were taken from the left anterior lobe of the rats from both experimental and control groups.Paraffin sections were prepared from materials fixed in:(1)Susa or Stieve's for Heidenhain's Azan staining;(2)Regaud's for Bensley's acid fuchsin-methyl green staining of mitochondria;(3)cold Carnoy's for methyl green-pyronin staining of ribo- nucleic acid (RNA),with control sections extracted with 20% perchloric acid or 0.17 M NaCl at 56℃ for 2 hrs.;(4)Carnoy's for Yakovlev's ((?)) and Barrnett- Seligman's methods for protein-bound SH-group,with control sections blocked with 0.1M monoiodoacetic acid;(5)Formol-alcohol for Hotchkiss's periodic acid-Schiff's (PAS) reaction of glycogen,with control sections treated with salivary digestion;(6)cold ace- tone for alkaline phosphatase (ALP);and (7) 10% neutral formalin for frozen sections and Sudan Ⅲ testing for fats. After CCl_4-intoxication,during the development of cirrhosis,most of the parenchymal cells degenerated and finally became necrotic.Free fat droplets and hyperplastic fibrous tissues accumulated in the necrotic regions.The liver lobules thus were destroyed and blood sinuses were engorged with blood cells.Surviving cells showed compensative hyperplasia and were isolated into nodules by the developing fibrous tissues.Cirrhosis developed in all the rats treated with CCl_4. The parenchymal cells showed a progressive changes during the development of cirrhosis.There was an increase of fat droplets and a decrease of the number of mito- chondria in the cytoplasm.The mitochondria fragmented into granules or fused in a mass.There was a progressive decline in the content of RNA and glycogen,but an increase of the ALP activity.Those cells showing compensative hyperplasia enlarged in size,and their number of mitochondria,RNA and glycogen content progressively in- creased. Small-sized cells appeared in the necrotic regions among the fat droplets,necrotic cells and fibrous tissues.They decreased in number as the fibrous tissue developed. These cells contained mitochondria and showed positive reactions of PAS,RNA and ALP.Their origin was discussed and their relation to the formation of collagen fibers needs further investigation.

Background and purpose:Recent researches have shown that Secreted Protien Acidic and Rich in Cysteine (SPARC) was closely related to tumor genesis, tumor progression and tumor metastasis. SPARC was highly expressed in malignant melanoma, glioma, meningioma, bladder cancer, lung cancer and prostate cancer, etc. In this study we investigated SPARC expression in hepatocellar carcinoma (HCC) and its signifi cance. Methods:RT-PCR was used to detect SPARC mRNA expression in cancer tissue samples and their adjacent liver tissue samples from 62 patients with hepatocellar carcinoma and 30 normal liver tissue samples, respectively. And the differential protein expression of SPARC between these groups was analyzed by immunohistochemistry (IHC). Results:SPARC mRNA was highly expressed in HCC(14.0?3.6) and in the adjacent liver tissue (6.8?1.8); compared with low expression of 2.7?0.9 in normal liver tissue, there were signifi cant differences among the three groups (p=0.000). SPARC positively stained was found in 54 of 62 patients with HCC and 4 of 30 normal liver tissue, there was significant difference between these two groups (P=0.000). SPARC immunohistochemical score was 21.5?4.8 in the carcinoma group; 11.3?3.6 in paracarcinoma group and 5.7?1.8 in the normal group, there were also significant differences among the three groups (P=0.000). The expression of SPARC protein was significantly upregulated with the progress of Enmondson pathological classification. There was obviously differences between Ⅰvs Ⅱ(P=0.029), and Ⅱ vs Ⅲ Ⅳ(P=0.008). There was more SPARC expression in the patients with metastasis of HCC (26/27, 96.3%) than that without metastasis(23/35, 65.7%)(P=0.004). Conclusion:SPARC mRNA expression and its protein were related to HCC histological differentiation and metastatic lymph node; SPARC is helpful to clinical evaluation of HCC.

50 adult albino rats were divided randomly into 3 groups, i, e. partial hepatectomy group, sham-operation group and normal control group. Specimens of tail portion of pancreas were taken at 1, 2, 3, 6 and 14 days after operation respectively. Paraffin sections were made and stained with immunohistochemical PAP method to demonstrate insulin-containing B cells,C-peptide-containing B cells,glucagon-containing A cells and somatostatin-containing .D cells. The changes in hormone secretion activity of B, A and D cells have been observed during liver regeneration period. Based on the different amount of immunoreactive product, the endocrine activity of islet cells can be divided into 3 grades: +, ++, and +++. Percentage of each grade of islet cells was calculated and taken as the parameter of endocrine activity of islet cells. Insulin release activity of B cells increased after 1-3 days of operation, and insulin biosynthesis and/or storage enhanced 3-6 days after. Glucagon release activity of A cells raised gradually during 1-3 days after operation, and somatostatin released activity of D cells declined at the same time. After 6 days, endocrine activity of A cells returned to normal level, but release of somatostatin of D cells elevated. Endocrine activity of islet cells all recovered to normal level 14 days later. The changes of endocrine activity of B cells and A cells were closely related to and parallel with regeneration process of liver. The changes of D cells suggest that it probably be a reflection of regulation action on endocrine activities of B cells and A cells.

50 adult rats divided into 3 groups, i. e. partial hepatectomy, sham-operation and normal control groups. The submandibular glands (SMG) were removed at different intervals (1-14 days) after operation, and stained with histological and histochemical method. On the 1-3 days after operation, especially on 3 days, some changes were found in granular convoluted tubule (GCT) cells of SMG. The secretory granules in the GCT cells were decreased in number. The reactions of PAS, -SS- group, PTAH were weaker than those of the sham-operation group, but the reactions of content in tubule system of SMG were raised. The reaction of SDH was stronger than those of the sham-operation group and the activities of NE and AcP decreased. After 7 days all the changes gradually restored, except the activities of ASDH increased and NE decreased continuously. After 14 days, all the changes became nearly normal. In addition, the reactions of RNA, G6Pase and 5-Nase in GCT cells of hepatectomized rats did not show apparent changes. The results suggested that, the changes of SMG GCT cells following partial hepatectomy seem, to be closely related with regenerative process of liver.

Objective To investigate the indications and methods of retaining only portal vein for the hilar cholangiocarcinoma radical resection to improve the therapeutic efficacy. Methods Six patients with hilar cholangiocarcinoma underwent left hepatectomy and extrahepatic bile duct excision. Tumor invaded hepatic artery was resected and fight hepatic duct-jejunum anastomosis was made during July 2006 to December 2007. Results There was no mortality. Bile leakage developed in one case and was cured by drainage. No liver failure developed postoperatively. All 6 cases were followed up for 10 - 23 months, and all were still alive. Conclusions Retaining only portal vein for the treatment of hilar cholangiocarcinoma radical resection under definite indications is safe.

Background and purpose: The previous work of this study has showed that the treatment of liver cancer cells with emodin could induce endoplasmic reticulum (ER) stress and apoptosis. Given the cross-talk between ER stress and autophagy, this study aimed to investigate whether blockage of autophagy, a defense mechanism against environmental stress, could improve the killing effect of emodin on liver cancer cells. Methods: The CYTO-ID auto-phagy detection kit and Western blot were used to determine autophagy in liver cancer cells. After combined treatment with chloroquine (CQ) and emodin, cancer cell survival was analyzed by ATPlite assay and clonogenic assay. Apoptosis was detected by both flow cytometry analysis and Western blot. Results: Autophagy could be induced in liver cancer cells after treatment with emodin. Inhibition of autophagy significantly increased growth-inhibitory effect of emodin on both HepG2 and Huh7 cancer cells. The combination treatment with CQ and emodin promoted remarkable apoptosis in liver cancer cells, evidenced by the increase in the percentage of cells in sub-G1 phase and the higher expression lever of cleaved caspase-3. Conclusion: Therapeutically targeting autophagy is capable of enhancing cytotoxicity of emodin in liver cancer cell lines.

OBJECTIVETo analyze isofraxidin and its metabolites in rat plasma, bile, urine and feces after the oral administration of A. senticosus extracts.

METHODAfter the oral administration of 325 mg x kg(-1) of A. senticosus extracts, rat blood samples were collected at 1, 1.5, 2, 4, 6 h and bile samples were collected during 0-12 h. The above samples were prepared by SPE. Their urine and feces samples were collected during 0-12 h and 12-24 h. These samples were analyzed by UPLC-Q-TOF-MS and processed using Metabolynx XS.

RESULTM0 was detected in rat plasma, bile, urine and feces, isofraxidin glucuronide (M1) was detected in rat plasma and bile, which was first reported as the metabolite of isofraxidin.

CONCLUSIONIsofraxidin can be metabolized in the form of glucuronidation in vivo and evacuated in the form of isofraxidin.

Animals ; Chromatography, High Pressure Liquid ; Coumarins ; blood ; metabolism ; pharmacokinetics ; Drugs, Chinese Herbal ; metabolism ; pharmacokinetics ; Eleutherococcus ; chemistry ; Male ; Mass Spectrometry ; Rats ; Rats, Wistar ; Statistics as Topic

OBJECTIVE: To investigate and reveal the underlying mechanism of the effect of total saponins from Dioscoreae nipponica Makino (TSDN) on the arachidonic acid pathway in monosodium urate (MSU) crystal-induced M1-polarized macrophages. METHODS: M1 polarization of RAW264.7 cells were induced by 1 µ g/mL lipopolysaccharide (LPS). The methylthiazolyldiphenyl-tetrazolium bromide method was then used to screen the concentration of TSDN. MSU (500 µ g/mL) was used to induce the gouty arthritis model. Afterwards, 10 µ g/L TSDN and 8 µ mol/L celecoxib, which was used as a positive control, were added to the above LPS and MSU-induced cells for 24 h. The mRNA and protein expressions of cyclooxygenase (COX) 2, 5-lipoxygenase (5-LOX), microsomal prostaglandin E synthase derived eicosanoids (mPGES)-1, leukotriene B (LTB)4, cytochrome P450 (CYP) 4A, and prostaglandin E2 (PGE₂) were tested by real-time polymerase chain reaction and Western blotting, respectively. The enzyme-linked immunosorbent assay was used to test the contents of M1 markers, including inducible nitric oxid synthase (NOS) 2, CD80, and CD86. RESULTS: TSDN inhibited the proliferation of M1 macrophages and decreased both the mRNA and protein expressions of COX2, 5-LOX, CYP4A, LTB4, and PGE₂ (P<0.01) while increased the mRNA and protein expression of mPGES-1 (P<0.05 or P<0.01). TSDN could also significantly decrease the contents of NOS2, CD80, and CD86 (P<0.01). CONCLUSION TSDN has an anti-inflammation effect on gouty arthritis in an in vitro model by regulating arachidonic acid signaling pathway.
Uric Acid/metabolism* ; Arachidonic Acid/metabolism* ; Dioscorea ; Arthritis, Gouty ; Lipopolysaccharides ; Saponins/pharmacology* ; Macrophages ; Signal Transduction ; RNA, Messenger/metabolism*