Objective To study the structure of spliced variants of hepatitis B virus (HBV) genomes and elucidate their potential pathogenicity. Methods Amplified the spliced variants of hepatitis B virus genomes by mean of PCR from the serum of the patients with chronic hepatitis B, sequenced and analysis the characteristics of such genomes. Results 10 different types of the spliced variants of hepatitis B virus genomes were obtained with the molecular weight ranging from 765 bp to 2039 bp. There were 6 splicing donor sites and 6 accepter sites, respectively in HBV genomes. All spliced variants showed one or more deletions in the regions coding for core, preS1, preS2 and surface protein, while retained thepathogenic X gene and cis elements which were essential for viral replication and packaging. Conclusions Spliced variants of hepatitis B virus genomes were commonly detected in the serum from chronic Hepatitis B patients, the characteristic structure of such variants implied that they might closely co-related with the pathogenicity of HBV.

ObjectiveTo investigate the effect of Dendrobium officinale polysaccharides （DOP） in reversing carbon tetrachloride （CCl₄）-induced hepatic fibrosis （HF） in rats via the Notch signaling pathway and evaluate the therapeutic effect of DOP by ultrasound elastography. MethodFifty-six male SD rats were randomized into normal， model， colchicine （1×10^-4 g·kg^-1）， Fuzheng Huayu powder （0.45 g·kg^-1）， and low-， medium-， and high-dose （0.05， 0.1， 0.2 g·kg^-1） DOP groups （n=8）. The rats in the model group and each treatment group were injected subcutaneously with a mixture of CCl₄-olive oil （2∶3） once every 3 days for 10 weeks. After 6 weeks of modelling， the rats were administrated with corresponding drugs once a day for 4 weeks. Hematoxylin-eosin （HE） staining and Masson staining were employed to observe the pathomorphological changes of the liver tissue. An automatic biochemical analyzer was used to measure the serum levels of alanine aminotransferase （ALT）， aspartate transaminase （AST）， alkaline phosphatase （ALP）， and total bile acids （TBA）. Enzyme-linked immunosorbent assay （ELISA） was employed to measure the serum levels of hyaluronic acid （HA）， laminin （LN）， type Ⅲ precollagen （PC-Ⅲ）， and type Ⅳ collagen （Col-Ⅳ）. The mRNA and protein levels of α-smooth muscle actin （α-SMA）， Notch1， Jagged1， and Hes1 in the liver tissue were determined by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot， respectively. The Young's modulus （YM） of the rat liver was measured by acoustic radiation force impulse （ARFI） elastography before and after treatment. Then， the correlations of YM with the serum levels of HA， LN， PC-Ⅲ， and Col-Ⅳ and the protein levels of α-SMA and Notch1 signaling pathway-related factors in the liver tissue were analyzed. ResultCompared with the normal group， the model group showed disordered arrangement of liver cell cords， obvious infiltration of inflammatory cells， appearance of a large number of fat vacuoles， and fibrous proliferation， elevated levels of ALT， AST， TBA， ALP， HA， LN， PC-Ⅲ， and Col-Ⅳ in the serum， and up-regulated mRNA and protein levels of α-SMA， Notch1， Jagged1， and Hes1 in the liver tissue （P<0.01）. Compared with the model group， drug interventions alleviated the pseudolobule formation and the collagen deposition in confluent areas. Except that the serum level of ALT in the low-dose DOP group had no significant changes， drug interventions， especially high-dose DOP， lowered the levels of ALT， AST， TBA， ALP， HA， LN， PC-Ⅲ， and Col-Ⅳ in the serum and down-regulated the mRNA and protein levels of α-SMA， Notch1， Jagged1， and Hes1 in the liver tissue （P<0.05， P<0.01）. The results of ARFI and correlation analysis showed that the YM of the liver tissue was increased in the model group （P<0.01） compared with that in the normal group， Compared with the model group， drug interventions decreased YM （P<0.01）. YM was positively correlated with the expression levels of HA， LN， PC-Ⅲ， α-SMA， Notch1， Jagged1， and Hes1s （r=0.754， 0.734， 0.801， 0.885， 0.896， 0.757， and 0.800， respectively， P<0.01）， and it had a moderate correlation with Col-Ⅳ （r=0.688， P<0.01）. ConclusionDOP can reverse HF by down-regulating the Notch1/Jagged1/Hes1 signaling pathway. YM can be used as an indicator in the assessment of the efficacy of DOP against HF.

OBJECTIVE To establish a high-performance liquid chromatography method for the simultaneous determination of 14 components including geniposide, mangiferin, baicalin, berberine hydrochloride, wogonoside, baicalein, aloe-emodin, rhein, wogonin, emodin, praeruptorin A, chrysophanol, physcion and praeruptorin B in Qingfei Yihuo tablets. METHODS Titank C18 (250 mm×4.6 mm, 5 μm) column was used; 0.1% phosphoric acid solution(A)-acetonitrile(B) was used as the mobile phase with gradient elution; detection wavelengths: geniposide at 238 nm, aloe-emodin, rhein, emodin, chrysophanol, physcion and mangiferin at 254 nm, berberine hydrochloride at 265 nm, baicalin, wogonoside, baicalein, and wogonin at 280 nm, praeruptorin A and praeruptorin B at 321 nm. The flow rate was 1.0 mL·min−1 and the column temperature was 35 ℃; the injection volume was 10 μL. RESULTS The linear ranges of geniposide, mangiferin, baicalin, berberine hydrochloride, wogonoside, baicalein,aloe-emodin, rhein, wogonin, emodin, praeruptorin A, chrysophanol, physcion and, praeruptorin B were 4.96−223.17, 0.84−42.22, 18.76−938.16, 4.46−223.17, 4.86−243.10, 1.59−79.32, 0.76−38.17, 1.03−51.49, 1.59−79.40, 1.21−60.72, 1.80−90.06, 0.91−45.48, 1.04−51.83 and 0.86−43.23 μg·mL−1, with r all ≥ 0.999 9. The RSDs of instrument precision, stability and reproducibility tests were <3%, and the average recoveries in sample(n=6) were >90%, with the RSDs <3%. CONCLUSION The method is simple and reproducible and can provide a scientific basis for improving the quality standard of Qingfei Yihuo tablets.

Objective To explore the change of complications and inflammatory mediators in patients at nutritional risk after intervention with fish oil after major abdominal surgery.Methods Totally 60 patients who had received extrahepatic bile duct exploration ± left lobectomy and radical surgeries for their gastrointestinal tumors in our department from September 2010 to March 2011 were enrolled in this study.Their Nutritional Risk Screening 2002 scores were ≥3 points the day after surgery.These 60 cases patients were randomly divided into fish oil group (n =30) and the control group (n =30).Both groups were treated with total parenteral nutrition isocaloric and isonitrogenous energy emulsion intralipid of 3 Ls' bags after surgery.Aslo,they were treated with electrolyte,vitamins and trace elements equally,which were adjusted according to laboratory tests.The application of parenteral nutrition was no less than 5 days,and the other treatment was provided routinely.Surgeries were performed under general anesthesia in all patients.Urinary catheter was uniformly placed preoperative,and was removed within 24 hours after surgery.The degree of surgical trauma was scored on the operative day.Fasting venous blood samples were collected for measuring the fasting interleukin (IL)-6 and tumour necrosis factor-α (TNF-α) one day before surgery and one and six days after surgery.The relevant complications were recorded.Two patients in the fish oil group withdrew from the study on the third post-operative day.Results TheIL-6 levels were (10.65 ± 4.24),(29.45 ± 9.39),and (13.37 ± 6.99) ng/Land the TNF-α levels were (2.47 ± 1.16),(23.05 ± 11.43),and (6.05 ± 2.97) ng/L in the fish oil group one day before surgery and one and six days after surgery.In the control group,in contrast,the IL-6 levels were (11.17 ± 4.67),(25.10 ± 10.13),and (17.38 ± 7.13) ng/L and the TNF-α levels were (2.70 ± 1.63),(22.11 ± 12.54),and (8.93 ± 3.61) ng/L at the corresponding time points.Comparisons of the IL-6 and TNF-α between the fish oil group and control group showed no significant difference one day before surgery and one day after surgery (P =0.787,P =0.206,P =0.983,P =0.852).Comparison of the IL-6 between the one day and six days after surgery also had no statistical significance (P =0.101).However,the decrease of IL-6 was significantly larger in the fish oil group than in the control group from day 1 to day 6 after surgery (P =0.036).The post-operative levels of TNF-α were significantly different between these two groups (P =0.024).Complications were noted in 3 cases (2 cases of infectious complications) in the fish oil group and in 6 cases in the control group (5 cases of infectious complications) (P =0.002),in which the inffectious complications were also statistically significant (P =0.001).Conclusions Patients at nutritional risks after major abdominal surgeries have higher TNF-α and IL-6 levels.Fish oil can notably reduce the TNF-α level in these patients.In addition,fish oil decreases the surgical complications,in particular the infectious complications.

ObjectiveTo investigate the effect of Dendrobium officinale polysaccharide （DOP）on CCl₄-induced hepatic fibrosis（HF）and its mechanism. MethodsA total of 56 male SD rats were randomly divided into seven groups： normal group（NG），model group（MG），colchicine group（CG， 0.1 mg/kg）， Fuzheng Huayu group（FG， 0.45 g/kg），low-dose DOP group（LDG， 0.05 g/kg），middle-dose DOP group（MDG， 0.1 g/kg）and high-dose DOP group（HDG，0.2 g/kg），with 8 rats in each group. HF rat model was established by subcutaneous injection with 40% CCl₄ olive oil mixture， every 3-day for 10 weeks. At the end of the sixth week， the drug groups were treated with colchicine， Fuzheng Huayu and DOP solution by gavage respectively， once a day for 4 weeks. NG and MG groups were similarly handled with an equal amount of 0.9 % normal saline. Liver histopathology was detected using hematoxylin-eosin （HE）， Masson and Sirius red staining； blood biochemistry was tested for liver function and four indicators of HF； RT-qPCR and Western Blot were used to measure the expression of α-SMA， Col-I， E-cadherin， and ZEB1 genes and proteins in the liver tissues of rats， respectively. ResultsHE， Masson， and Sirius red staining showed that the liver tissue of MG rats had typical pathologic features of HF， and the degree of HF was alleviated in LDG， MDG， and HDG rats， respectively. Liver function test results showed that the serum AST， TBIL， and AKP levels were significantly lower in LDG， MDG， and HDG， compared with those of the MG （P < 0.05 or < 0.01）. Meanwhile， ALT levels in serum deceased remarkably except in LDG （P < 0.05 or < 0.01）. The four results of HF showed that the serum HA， LN， PC-Ⅲ， and COL-Ⅳ levels in LDG， MDG， and HDG rats were significantly decreased compared with those of the MG （P < 0.05 or < 0.01）. The relative expressions of α-SMA， COL-I， and ZEB1 genes and proteins were significantly decreased in the liver tissues of LDG， MDG， and HDG （P < 0.05 or < 0.01）， and the relative expression of E-cadherin gene and protein increased （P < 0.05 or < 0.01）. In addition， the expressions of HA， α-SMA， COL-I， ZEB1 and E-cadherin were dependent on the dose of DOP. ConclusionDOP alleviated the degree of CCl₄ induced HF in rats by inhibiting the epithelial-mesenchymal transition in liver tissue.

Objective:To investigate the differences of chemical constituents of Salviae Yunnanensis Radix et Rhizoma and its closely related species,and to provide reference for the clinical application of Salviae Yunnanensis Radix et Rhizoma. Method:Fourier transform infrared (FTIR) spectroscopy combined with principal component analysis (PCA),cluster analysis (HCA) and partial least squares discriminant analysis (PLS-DA) were used to study the differences of components between Salviae Yunnanensis Radix et Rhizoma and its closely related species,and high performance liquid chromatography (HPLC) was used to compare the differences of water-soluble components between them. Result:There were some differences between Salviae Yunnanensis Radix et Rhizoma and its closely related species acrcording to FTIR spectroscopy and HPLC fingerprint,especially the water-soluble polar components were more abundant in Salviae Yunnanensis Radix et Rhizoma than other species. The chemical components of Salvia trijuga,S. przewalskii and S. bulleyana were more similar in terms of their genetic relationship. The result showed that the Salviae Yunnanensis Radix et Rhizoma and its closely related species can be clearly distinguished by FTIR combined with chemometrics method. Conclusion:Compared with its closely related species,Salviae Yunnanensis Radix et Rhizoma has a unique chemical composition,which has great therapeutic potential for blood stasis. The FTIR combined with chemometrics model can be used for rapid identification of Salviae Yunnanensis Radix et Rhizoma and its closely related species.

Keratitis-ichthyosis-deafness (KID) syndrome has genetic heterogeneity, and the clinical manifestations of some patients may overlap with Clouston syndrome. A 34-year-old female patient came to our department with a complain of “sparse hair, rough skin, photophobia and deafness for more than 30 years.” We found that the proband and two other family members (57-year-old mother and 4-year-old daughter) had similar clinical manifestations:systemic hair loss, generalized skin hyperkeratosis, especially in the metacarpophalangeal area. Subungual hyperkeratosis, finger/toenail dystrophy, as well as photophobia and epiphora. According to the investigation, one of the family members also had similar clinical manifestations (grandfather of the proband) and he’s died. The other three members of the family had no hearing impairment, and all patients had typical nail dystrophy, hair loss and palmoplantar hyperkeratosis, similar like as seen in Clouston syndrome, so we suspected to diagnose the case as Clouston syndrome. However, after genetic testing, it was found that the proband, his mother and daughter all had p.Asp50Asn heterozygous mutations in the GJB2 gene, and no mutation was detected in GJB6. The modified diagnosis was KID syndrome.

This study expounded the great impacts of excess energy intake and oxidative inflammation on national health in China and put forward the pathological mirror-image hypothesis of the relationship between obesity and oxidative inflammation. On this basis, an ideological framework was constructed to deal with oxidative stress and oxidative inflammation centered on the development of natural antioxidant products. The study contained the following four parts: the ways to improve national health in China; the promotion of cognition to oxidative inflammation to improve national health in China; prospects for the prevention and treatment of oxidative inflammation in China; the reason why Northwest Yunnan is a good place to develop high-quality natural antioxidants.
China ; Energy Intake ; Humans ; Inflammation ; Oxidation-Reduction ; Oxidative Stress

Malignant tracheoesophageal fistula (MTEF) is pathological communication between the respiratory tracts such as the trachea or bronchia and the esophagus because of malignant tumor dissemination through them.Radiography is an important adjunctive technology in the diagnosis of MTEF,and the location and size of fistula often need the further diagnosis of bronchoscopy and esophagoscopy.The patients are often with an unfavourable prognosis once developed MTEF,and are treated usually with the aim of symptom palliation and life quality improvement.The individual treatment includes esophageal stenting,esophageal exclusion and esophagus bypass,fistula exclusion and repair,radiotherapy and others effective therapy according to the patients condition.These therapies will prolong the life span and improve the life quality of patients.MTEF is not absolute contraindication for chemoradiotherapy.Despite its acute toxicity,this concurrent chemoradiothe rapy protocol appears feasible and effective at closing esophageal malignant fistula,especially in patients in a good general condition and without metastasis.

Objective To assess the effects of hepatitis B virus(HBV) large envelope protein (LHB) on the apoptosis of HepG2 cells and explore the possible mechanism by proteomic approaches.Methods LHB gene was cloned into pShuttle-IRES-hrGFP-1,and the recombinant adenovirus either barboring LHB(Ad-LHB) or empty vector(Ad-GFP) were separately generated.Annexin V-FITC apoptosis detection kit,JC-1 mitochondrial membrane potential assay kit and propidium iodide(PI) staining kit were employed combined with flow cytometry to detect the apoptotic cells infected with Ad-LHB or control of Ad-GFP.The cellular proteins were collected after infection of HepG2 cells by Ad-LHBs or Ad-GFP,and a total of 600 μg proteins were submitted to two-dimensional gel electrophoresis(2-DE) and stained with R350.The gel images were captured by ImageScanner Ⅱ Imaging System,the differentially expressed proteins were identified by ImageMaster 2D Platinum analysis software and picked up by Ettan Spot Picker.After enzyme digestion,the protein samples were analyzed by MALDI-TOF-TOF MS.Results HepG2 cells infected with Ad-LHB were much more prone to apoptosis.There were thirty nine differentially expressed proteins were determined by 2-DE between HepG2 cells infected with Ad-LHB and Ad-GFP,and they were identified ultimately and categorized into thirty three kinds of proteins by MALDI-TOF-TOF MS.Among these proteins,nine were found to be closely related to cell apoptosis,in which CAPN2,eIF3K and PPP2CB were higher expressed in Ad-LHB infected HepG2 cells,and SERPINH1,LASP1,PRDX1,DHRS2,LDHA and PS-MA4 were lower expressed in Ad-LHB infected HepG2 cells.Conclusion LHB could induce apoptosis of HepG2 cells,and several apoptosis-related proteins participated in this process.