Association between liver fibrosis and gut microbiota based on "harmonizing liver and spleen" / 临床肝胆病杂志

Liver fibrosis is the leading cause of the morbidity and mortality of liver diseases worldwide and has become one of the key issues in the research on chronic liver diseases among domestic and foreign scholars. An increasing amount of evidence has shown that gut microbiota dysbiosis plays a crucial role in the development and progression of liver fibrosis, and improvement of gut microbiota dysbiosis has become a new target for anti-fibrotic treatment. At present, traditional Chinese medicine has attracted much attention in improving gut microbiota. Studies have shown that "harmonizing liver and spleen" plays an important role in reducing fibrosis degree and delaying the progression of liver fibrosis. Therefore, based on the theory of "harmonizing liver and spleen", the achievements in liver-gut axis and gut microbiota by modern molecular biological techniques can provide a theoretical basis for the treatment of liver fibrosis from "harmonizing liver and spleen" in clinical practice, as well as new directions and ideas for anti-fibrotic treatment with traditional Chinese medicine.

An investigation of hepatitis D virus infection among patients with chronic hepatitis B virus infection in some regions of China / 临床肝胆病杂志

Objective To investigate the prevalence of hepatitis D virus (HDV) infection among patients with chronic hepatitis B virus (HBV) infection in some regions of China. Methods Serum samples were collected from 3 131 patients with chronic HBV infection in 10 provinces, cities, and autonomous regions of China from March 2021 to June 2022, and anti-HDV IgG ELISA was used for the detection of all serum samples. Nested reverse transcription-polymerase chain reaction (nRT-PCR) was used to detect HDV RNA in anti-HDV IgG-positive samples, and the nRT-PCR amplification products of HDV RNA-positive samples were sequenced and analyzed to determine HDV genotype. The clinical features of anti-HDV IgG-positive patients were analyzed. The Mann-Whitney U rank sum test was used for comparison of continuous data between two groups, and the chi-square test or the Fisher's exact test was used for comparison of categorical data between two groups. Results The positive rate of anti-HDV IgG in the 3 131 patients with chronic HBV infection was 0.70% (22/3 131), and that in the patients with chronic HBV infection in Inner Mongolia Autonomous Region, Xinjiang Uygur Autonomous Region, Beijing, and Hunan Province was 1.81% (16/886), 0.88% (2/226), 0.28% (2/708), and 1.00% (2/200), respectively; the patients with chronic HBV infection in Inner Mongolia Autonomous Region had a significantly higher positive rate of anti-HDV IgG than those in Beijing ( P =0.004), and there was no significant difference between the other regions ( P > 0.05). Clinical features of the patients with chronic HBV infection in Inner Mongolia Autonomous Region showed that compared with the anti-HDV IgG-negative group, the anti-HDV IgG-positive group had a significantly higher proportion of patients with Mongol nationality ( P =0.001), abnormal alanine aminotransferase ( P =0.007), or antiviral treatment ( P =0.029), as well as a significantly lower median HBV DNA level ( P =0.030). A total of 19 HDV RNA-positive samples were identified, all of which had HDV genotype 1. Conclusion The prevalence rate of HDV varies greatly across different regions of China, with a higher prevalence rate of HDV in patients with chronic HBV infection from Inner Mongolia Autonomous Region. HDV genotype 1 is the predominant genotype in some provinces and cities of northern China.

Clinical value of endoscopic release therapy for fecal impaction in colorectal diverticulum / 中华消化内镜杂志

Objective:To evaluate the clinical value of endoscopic release therapy for fecal impaction in the colorectal diverticulum.Methods:Data of patients with fecal impaction in the colorectal diverticulum who received endoscopy in Shantou Central Hospital from January 2018 to September 2020 were included in this study. Among them, 85 patients treated with endoscopic release therapy were assigned to the observation group (2 patients were excluded from the observation group due to acute appendicitis), and 43 patients receiving no treatment were assigned to the control group. The relief of abdominal symptoms was used as an index to evaluate the clinical value of endoscopic release therapy for fecal impaction in the colorectal diverticulum.Results:In the observation group, 42.2% (35/83) were successfully released at one time. The successful comprehensive measures accounted for 25.0% (12/48) of the first release failure, and the total success rate was 56.6% (47/83). There were no complications related to endoscopy in the observation group. One week after the treatment, patients in the observation group were followed up by telephone. Among the 45 patients who were successfully released, positive symptoms of 30 patients disappeared or significantly improved with the effective rate of 90.9% (30/33). Among the 38 patients who failed to release the fecal impaction, 19 had positive symptoms and 16 improved in varying degrees with the effective rate of 55.2% (16/29). The overall effective rate of the observation group was higher than that of the control group [55.4% (46/83) VS 7.0% (3/43)], showing significant difference( χ2=23.354, P<0.01). The effective rate were significant differences in the successful release group [65.2% (30/46)], unsuccessful release group [29.7% (11/37)] and the control group [7.0% (3/43), χ2=33.792, P<0.01]. By pairwise comparison, the effective rate of the successful release group was the highest, followed by the unsuccessful release group, and that of the control group was the lowest with significant difference ( P<0.017). Conclusion:The endoscopic release therapy for fecal impaction in the colorectal diverticulum is relatively simple, which can relieve and reduce related symptoms, avoid complications, missed diagnosis and misdiagnosis, and show definite curative effects. When colorectal diverticulum with fecal impaction is found in the process of endoscopy, it is of great practical significance to release the incarcerated feces by means of different methods.

Significance of alternative splicing in drug resistance of chronic myeloid leukemia / 中国肿瘤临床

Alternative splicing (AS) is a process by which the transcriptome diversity, and thereby the proteome diversity, is augment-ed by splicing or joining together different parts of the pre-mRNA in eukaryotic cells . AS at different splice sites is regulated by multi-ple cis-acting elements and trans-acting factors. Chronic myeloid leukemia (CML) is a clonal myeloproliferative disease in which there is a translocation between the long arms of chromosome 9 and chromosome 22, represented as t(9;22) (q34;q11). This translocation results in the formation of a BCR-ABL fusion gene. Hence it is not surprising that resistance to tyrosine kinase inhibitors, which inhibit BCR-ABL activity, has become a critical problem in the clinical treatment of CML. Using second generation high-throughput sequencing technology, it has been found that AS abnormalities are closely related to the occurrence, progression, drug resistance, and immune escape of CML. This paper reviews the research related to AS and CML resistance and investigates the potential causes of CML resis-tance. Drug resistance mechanisms and potential therapeutic targets are also reviewed.

Effect of narrow-band imaging (NBI) combined with magnifying endoscopy in early diagnosis of gastric cancer / 中国医师杂志

Objective To explore the effect of nanow-band imaging (NBI) combined with magnifying endoscopy (ME) in early diagnosis of gastric cancer,and to observe whether it is helpful for inexperienced endoscopists to diagnose.Methods From April 2015 to October 2017,a total of 480 lesions were selected as research objects which were detected in the normal white light endoscopy (WLE).Pathological examination was used as the gold standard for diagnosis.3 experienced doctors were selected to read the endoscopy films.40 intern endoscopy doctors were selected to read the endoscopy filins at the same time after simple training.80 WLE images and narrowband imaging technology combined with magnifying endoscopy (ME-NBI) images were randomly selected to hold a film reading meeting.Real-time voting was used to judge the nature of lesions.Results The accuracy,sensitivity,specificity false positive rate and false negative rate of ME-NBI were 97.9％ (470/480),96.8％ (120/124),98.3％ (350/356),1.7％ (6/356),3.2％(4/124).And WLE values were 91.7％ (440/480),80.6％ (100/124),89.9％ (320/356),10.1％ (36/356),19.4％ (24/124),respectively.The diagnostic accuracy,sensitivity and specificity of ME-NBI were significantly higher than those of WLE (x2 =18.989,16.104,15.859,P≤0.05),and the false positive rate and false negative rate were significantly lower than those of WLE (x2 =22.772,16.104,P ≤ 0.05).The diagnostic results of ME-NBI of 40 endoscopy interns were significantly better than those of WLE (t =11.467,16.188,P ≤0.05).The diagnostic results of NBI by 40 interns were significantly better than that of WLE (t =11.467,16.188,P ≤ 0.05).Conclusions The accuracy of ME-NBI in screening early gastric cancer is satisfactory,and it has certain diagnostic value for inexperienced doctors.

IL-6 /STAT-3 signaling pathway regulates the expression of high mobility group proteins 1 in intestinal mucosa of rats with sepsis / 中华急诊医学杂志

Objective To explore whether the IL-6/STAT3 signaling pathway regulate the expression of high mobility group proteins1 (HMGB1) in intestinal mucosa of rats with sepsis through the cecum ligation puncture (CLP). Methods One hundred and twenty male SD rats were randomly(random number) divided into three groups: sham operation group (group S, n=40), CLP group(group C, n=40) and anti-IL-6 monoclonal antibody group (group T, n=40). Rats in group S only received the simple laparotomy;Rats in group C and group T were established as a rat model of sepsis using CLP; rats in group T received the intraperitoneal injection of anti-IL-6 monoclonal antibody at 1 h after CLP, while the same volume of sodium lactate ringer's solution was injected to rats in group S and group C. Ten rats in each group were sacrificed at 3, 12, 24 and 48 h, respectively, and intestinal mucosa specimens were collected for pathological examinations by HE staining. The protein expression of HMGB1 and IL-6 were detected by immunohistochemistry, STAT3-protein by Western blot.and the levels of diamine oxidase (DAO) and D lactic acid in plasma by spectrophotometric. Results Rats in group C and group T showed obvious intestinal damage to different degrees, significantly higher intestinal mucosa pathological scores and plasma levels of DAO and D-lactic acid compared with rats in group S (P<0.05). The protein expression of IL-6, HMGBl and p-STAT3 of intestinal mucosa in group C and group T also significantly increased compared with that in group S (P<0.05). The intestinal mucosa pathological score, plasma levels of DAO and D-lactic acid and protein expression of IL-6, HMGBl and STAT3 were decreased in group T compared with those in group C (P<0.05). The intestinal mucosa pathological scores were positively correlated with the protein expression of IL-6 and HMGB1 at 12, 24, and 48 h, respectively. Conclusions IL-6 and HMGBl were involved in the intestinal injury of septic rats. IL-6/STAT3 signaling pathway could up-regulate the expression of HMGB1 in intestinal mucosa of septic rats.

Study on Medication Rules of Professor ZENG Bin-fang’s Treatment for Chronic Hepatitis B ;Based on Complex Network Analysis Method / 中国中医药信息杂志

Objective To analyze the medication rules of Professor ZENG Bin-fang’s treatment for chronic hepatitis B (CHB); To provide references for clinic. Methods Research data came from patients with CHB in outpatient and hospitalization in Department of Hepatology, Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous Region. In this study,data mining methods, such as complex network analysis and dot-mode mutual information, were utilized to conduct analysis on the syndromes, symptoms, treatment and medication of included patients, as well as the mutual relations. Results The study collected clinical data of 132 patients with CHB, who accepted 277 times of examination,and 277 prescriptions were included. High-frequency symptoms included:burnout and weakness, abdominal distension, hypochondriac pain, and lower limb edema; Common syndromes included:stagnation of liver qi and spleen deficiency, retention of damp-heat in the interior, liver and gallbladder damp-heat, liver qi stagnation, and qi-stagnation and blood stasis, etc., successively, including 126 times of examination and accounting for 45.5%of all cases. Common therapeutic methods involved in soothing liver and strengthening spleen, eliminating dampness and heat, nourishing liver and kidney, regulating vital energy and promoting blood flow and activating blood and activating spleen, successively. Dot-mode mutual information analysissyndrome-symptom relation:the differentiation point of stagnation of liver qi and spleen deficiency included burnout and weakness, thirst,dull pain of right lateral thorax, anorexia, and abdominal distension;syndrome-therapy relation:stagnation of liver qi and spleen deficiency contained the main therapeutic method of nourishing liver and strengthening spleen;syndrome-medicine relation: Bupleuri Radix, Codonopsis Radix, Scutellariae Radix, Angelicae Sinensis Radix, Paeoniae Radix Rubra,Glycyrrhizae Radix et Rhizoma Praeparata cum Melle, Pinellinae Rhizoma Praeparata, raw Rehmanniae Radix, and Cyperi Rhizome. Analysis of complex network analysis showed that the core formula for stagnation of liver qi and spleen deficiency was Xiaochaihu Decoction and Siwu Decoction. Conclusion The treatment thinking of Professor ZENG Bin-fang for CHB pays attention to entire concept, attaches importance to Zang-fu transmission, regards“nourishing liver, soothing liver and strengthening spleen”as the therapeutic principle, together with nourishing yin, softening and resolving hard mass, clearing heat-toxin and eliminating dampness.

The predictive values of three noninvasive indices in diagnosis of liver fibrosis in patients with chronic hepatitis B: a comparative study / 临床肝胆病杂志

ObjectiveTo investigate the values of FibroTouch, FIB-4 index, and aspartate aminotransferase-to-platelet ratio index (APRI) in the diagnosis of liver fibrosis in patients with chronic hepatitis B (CHB). MethodsA total of 148 patients with CHB who visited Traditional Chinese Medicine Hospital of Xinjiang Uygur Autonomous Region and underwent liver biopsy from September 2013 to May 2015 were enrolled and divided into groups according to fibrosis stage. All the patients underwent blood biochemical examination, routine blood tests, and FibroTouch measurement. Then FIB-4 and APRI were calculated, and liver stiffness was recorded. The receiver operating characteristic (ROC) curve was used to calculate the area under the ROC curve (AUC) and determine the cut-off value, sensitivity, and specificity. Chi-square test was used for comparison between two groups, and the Pearson rank correlation analysis was also performed. ResultsFibroTouch, APRI, and FIB-4 were well correlated with fibrosis stage (r=0.628, 0.486, and 0482, respectively, all P＜0.01). In the marked liver fibrosis (≥S2) group and liver cirrhosis (S4) group, FibroTouch had the best diagnostic performance, with AUCs of 0.84 and 0.93, respectively, followed by APRI, which had AUCs of 0.79 and 0.87, respectively; FIB-4 index had the worst diagnostic performance, with AUCs of 0.77 and 0.84, respectively. In patients with a fibrosis stage of ≥S2 or S4, FibroTouch had a better diagnostic value than APRI and FIB-4 (Z=21.589, P＜0.001; Z=18.896, P＜0.001; Z=11.192, P=0.001; Z=16.891, P＜0.001), and APRI had a better diagnostic value than FIB-4 (Z=46.918, P＜0.001; Z=35.334, P＜0.001). ConclusionFibroTouch can accurately evaluate the presence of liver fibrosis and fibrosis degree and help most patients avoid invasive liver biopsy.

Mechanism of alternative splicing and its application in diagnosis and treatment of leukemia / 中华检验医学杂志

Alternative splicing is a process refering a pre-mRNA transforms to different mature mRNAs by different splicing sites combination, and then the mRNAs translate to various proteins.This process is regulated by a variety of cis acting elements and trans acting factors.Recent studies have shown that aberrant alternative splicing is prevalent in leukemia patients, and is closely associated with leukemic occurrence, development and chemotherapy resistance.This shows us that aberrant altemative splicing may be helpful to leukemia diagnosis and treatment.

Logistic Regression Analysis on Risk Factors of Northwest Dryness Syndrome Among Patients of Metabolic Syndrome in Xinjiang Uygur Autonomous Region / 中国中医药信息杂志

Objective To analyze risk factors of northwest dryness syndrome (NDS) among the patients of metabolic syndrome in Xinjiang Uygur Autonomous Region.Methods Totally 321 cases of metabolic syndrome in Kashgar, Hami and Urumqi were collected to filter 15 variables to calculate OR value and 95% CI, and evaluated by single factor and multi-factor analysis by applying conditional Logistic regression analysis.Results Single factor conditional Logistic regression analysis showed that high fat diet, addiction to hot diet, salty diet, smoking history and hypertension were the risk factors of NDS. The OR values of them were 0.387, 0.071, 0.033, 2.614 and 0.440, respectively (P<0.05). Multi-factor conditional Logistic regression analysis showed that smoking history, alcohol drinking history, high fat diet and hypertension were all entered into regression model. The OR values of them were 3.945, 4.334, 0.370 and 2.142, respectively (P<0.01).Conclusion Patients of metabolic syndrome with a high fat, salty and hot diet are suggested to adjust their diet habit by adding sour food, giving up smoking and drinking, and controlling their blood pressure. It will be helpful for them to adapt local climate and living environment, and reduce the attack rate of NDS.

Correlation Study on Uyghur Metabolic Syndrome and Northwest Dryness Syndrome in / 中国中医药信息杂志

Objective To analyze the correlation between Uyghur metabolic syndrome and northwest dryness syndrome in Kashgar prefecture, Xinjiang. Methods Case-control design was used to include a total of 120 cases from the First People’s Hospital of Kashgar Prefecture, Xinjiang. They were selected as the case group, and 120 cases without diseases of Western medicine were as the control group. The northwest dryness syndrome was statistically analyzed. Results The total integral of northwest dryness syndrome in case group was (11.49±0.89) and that in control group was (5.84±0.81), and the case group was higher than the control group with statistical significance (P<0.01). By comparing various accompanied symptoms, except for the fire & wind dryness syndrome of lung&heart&spleen as well as lung&stomach yin deficiency syndrome, all the other syndromes in case group were higher than those in control group (P<0.01). The attack rate of northwest dryness syndrome in case group was 24.17% (29/120), higher than that in control group (7.50%, 9/120), with significant difference (P<0.01). In the correlation analysis between the state of TCM syndrome and northwest dryness syndrome in case group, spleen yin deficiency syndrome of metabolic syndrome had the strongest correlation with damp accumulation syndrome of northwest dryness syndrome (P<0.01), followed by liver-qi stasis syndrome and heart&kidney yin deficiency syndrome (P<0.01). Conclusion Metabolic syndrome is often concomitant with northwest dryness syndrome in Kashgar prefecture, Xinjiang. Moreover, the severity of clinical symptoms of metabolic syndrome was associated with the state of northwest dryness syndrome, which provides clinical epidemiology basis for developing prevention and cure strategy of metabolic syndrome in Xinjiang from the perspective of“treating the different diseases with the same therapy”.

Preparation of Polyacrylamide Resin and Its Adsorption Properties on 2,4-Dichlorophenoxyacetic Acid / 分析化学

Polyacrylamide resin was synthesized via surface-initiated atom transfer radical polymerization ( SI-ATRP) method. Acrylic amide ( AM) was grafted onto the surface of the chloromethyl polystyrene resin via SI-ATRP in the CuBr/2, 2'-bipyridine ( Bpy) system as catalyst at room temperature. The compositions of polyacrylamide resin were determined by means of elementary analysis, FT-IR analysis and scanning electron microscopy ( SEM) . The adsorption properties, the parameters of kinetics and the thermodynamics of the resin were evaluated in details, respectively. As the results, adsorption capacity of 2,4-dichlorophenoxyacetic acid (2,4-D) increased with the initial concentrations of solution increasing at room temperature, and its highest adsorption capacity was 111. 0 mg/g with solution concentration of 8 mmol/L. Adsorption isotherm at room temperature was determined and modeled with Langmuir and Freundlich equations. The thermodynamic equilibrium functions were calculated to be ΔG0, hence, the adsorption was spontaneous, endothermic and entropy increasing. The kinetics fitted the pseudo-second-order well. The polyacrylamide-chloromethyl polystyrene (PAM-CMCPS) resin was used for the adsorption of 2,4-D in orange sample, and good results were obtained.

Effect of siRNA targeting HDAC1 gene on proliferation, apoptosis, histone acetylation, and histone methylation in gastric cancer cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of silencing histone deacetylases 1 (HDAC1) gene by RNA interference on the proliferation, apoptosis and histone modulation in gastric cancer MGC-803 cell line.</p><p><b>METHODS</b>The optimal segment targeting HDAC1 gene was designed and transfected into MGC-803 cells by Lipofectamine TM2000. HDAC1 mRNA and protein in the transfected cells were detected by RT-PCR and Western blotting, respectively. The growth inhibition of MGC803 cells was evaluated by MTT assay and the cell apoptosis was detected with TUNEL assay. The expression of Bcl-2, procaspase-9, procaspase-3, c-Myc, histone acetylation of H3, H4, and histone methylation of H3K9 was detected by Western blotting.</p><p><b>RESULTS</b>The siRNA targeting HDAC1HDAC1 markedly suppressed mRNA expression, inhibited cell proliferation and induced apoptosis of MGC-803 cells in a concentration manner. Transfection of the cells with HDAC1 siRNA at 0, 30, 60, and 120 nmol/L for 24 h resulted in a cell apoptotic rate of (4.8∓2.7)%, (18.5∓3.5)%, (41.4∓4.3)%, and (59.2∓5.5)%, respectively, and caused down-regulation of the expressions of Bcl-2, proCaspase9, proCaspase3 and c-Myc, upregulation of histone acetylation of H3, H4, and down-regulation of histone methylation of H3K9.</p><p><b>CONCLUSION</b>Silencing HDAC1 gene expression with HDAC1 siRNA can promote histone H3 and H4 acetylation and inhibit histone methylation of H3K9 to suppress the proliferation and induce apoptosis of gastric cancer MGC-803 cells.</p>

Changes of liver sinusoidal endothelial cells and basement membrane in early stage of liver fibrosis induced by carbon tetrachloride in rats / 中国病理生理杂志

[ABSTRACT]AIM:Toexplorethedevelopmentofhepaticsinusoidalcapillarizationintheearlystageofliverfi-brosis induced by carbon tetrachloride (CCl4) in rats.METHODS:Clean SD rats were randomly divided into normal con-trol group (group N, n=6) and liver fibrotic model group (group M, n=32).The rats in group N were intraperitoneal in-jected with saline and the rats in group M were intraperitoneal injected with CCl 4(2 mL/kg, twice a week for 4 weeks).At the end of the 3rd day and the 1st, 2nd and 4th weeks, all rats were killed and then the samples were collected .The patho-logical changes in the livers were observed by HE staining and Masson straining .The development of hepatic sinusoidal capillarization was observed by transmission electron microscopy (TEM) and immunohistochemical staining .The cell sur-face expression of vascular endothelium-associated marker CD31, collagen type Ⅳ(Col IV) and laminin (LN) was deter-mined.RESULTS:HE and Masson staining showed the formation of liver fibrosis after treatment with CCl 4 for 4 weeks. TEM showed that the fenestrate diameter of liver sinusoidal endothelial cells (LSECs) grew down, the fenestrate numbers of LSECs were decreased along with the development of liver fibrosis , and the consecutive basement membrane was formed at the end of the experiment .The expression of CD31 was significantly increased along with the development of defenestration , and the expression of Col IV and LN was significantly increased after the treatment with CCl 4 for 2 weeks and 4 weeks , re-spectively .CONCLUSION:The typical hepatic sinusoidal capillarization was detected in the early stage of liver fibrosis , and the deposition of LN in the liver sinusoidal walls was the mainly factor of formation of the consecutive basement mem -brane .

Meta-analysis of Randomized Controlled Trail of Traditional Chinese Medicine on Children with Mycoplasma Pneumonia / 中国中医药信息杂志

Objective To evaluate the effectiveness and safety of TCM treatment to children with mycoplasma pneumonia. Methods RCTs of trational Chinese medicine in treating children with mycoplasma pneumonia were retrieved. CNKI, Wanfang Database and CBMdisc were searched for Chinese literature, and PubMed, EMbase and Cochrane Library were searched for foreign literature. All searching time was up to May 25, 2012. Data were picked up by two system reviewers individually and evaluation standard of Cochrane was used to evaluate the quality of selected literature. Results A total of 64 RCTs including 6289 cases of children with mycoplasma pneumonia were analyzed. In the improvement of clinical symptoms, among comparison between traditional Chinese medicine and western medicine (RR=1.10, 95%CI:-1.0--1.18) and comparison between traditional Chinese medicine plus western medicine and western medicine (RR=1.14, 95%CI:1.10-1.18), curative effects of treatment group were superior to the control group, P<0.00;For the fever clearance time, among comparison between traditional Chinese medicine and western medicine (RR=-4.26, 95%CI:-8.15--0.37), comparison between traditional Chinese medicine plus western medicine and western medicine (RR=-0.97, 95%CI:-1.27--0.66), curative effects of treatment group were superior to the western medicine group. For the time of the disappearance of cough and pulmonary rales, and the basic X-ray absorption time, curative effects of treatment group were also superior to the control group. Untoward effects in western medicine group were obvious than that of traditional Chinese medicine group. Conclusion TCM could improve and shorten the duration of symptoms of children with mycoplasma pneumonia. However, it is still necessary to be verified with more multi-center, double-blind, large sample randomized controlled trials, since the literature which have been brought into related researches were with low quality.

Location of the binding position for HBV X protein functional interaction with cytochrome C oxidase III / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To identify the binding site position of the hepatitis B virus (HBV) X protein (HBx) functional interaction with the cytochrome C oxidase subunit III (COX III, a key regulator of mitochondrial function) by using a yeast two-hybrid system.</p><p><b>METHODS</b>Two fragments of HBx mutants (X1 1-72aa and X2 1-117aa) were amplified by PCR and inserted into the bait plasmid pAS2-1.The resultant mutant plasmids were transfected into yeast cells using the lithium acetate-method.PCR and gene sequencing were used to confirm that the mutant fragments were expressed properly in yeast cells.Western blotting was used to verify that the mutant proteins were translated accurately in the yeast cells.Filter assay was used to exclude autoactivated mutants.Hybridization in solid medium and beta-gal activity detection were used to determine the precise position of the binding site for HBx and COX III interaction.</p><p><b>RESULTS</b>The two mutant plasmids containing HBx 1-72aa and 1-117aa respectively were successfully constructed and the mutants were both properly expressed and translated in yeast cells; no autoactivated mutants were detected throughout the experimental process.The binding site of HBx and COX III was found to be encompass the amino acids 72 through 117 of HBx.</p><p><b>CONCLUSION</b>Amino acids 72 through 117 of HBx are the key domain of the HBx functional interaction With COX III; this domain may represent a useful target for molecular-based therapies to treat HBV-related diseases.</p>

Effects of rat interleukin-10 gene treatment on expression of collagen, MMP13 and TIMP1 in fibrotic rats / 中国免疫学杂志

Objective:To study the effects of rat interleukin-10 (rIL-10) gene treatment on the expression of collagen , matrix metalloproteinase 13(MMP13) and their specific inhibitors the tissue inhibitor of metalloproteinase 1(TIMP1) in porcine serum in-duced liver fibrosis rats then to explore the anti-fibrotic effect of rL-10.Methods:Thirty SD rats were divided into normal control and fibrosis model group.Normal control group (group C) was intraperitoneally injected with 0.5 ml normal sodium twice a week for 8 week, while the fibrosis model group was injected with equal volume of pig serum for 8 week.At the beginning of the 5th week, fibrosis model group was further randomly divided into a fibrosis model subgroup ( group M ) , rIL-10 gene treatment subgroup ( group I ) and empty vector control subgroup(group P).Rats in group C and M were injected with Ringer’s solution as a reagent control via the tail vein weekly, rats in group I were injected with the rIL-10 plasmid pcDNA3-rIL-10, and rats in group P were injected with empty vector pcDNA3.All rats were sacrificed at the end of 8th week, and the liver tissue samples were collected to observe deposition of collegan in liver tissue by sirius red staining and detected the expression of MMP 13 and TIMP1 in the liver tissue by SP immunohistochemistry .Re-sults:Sirius red staining showed that the area of the collegan deposition was dramatically increased in fibrosis model subgroup and emp -ty vector control subgroup compared with the normal control group , and the area of the collagen deposition was dramatically decreased in rIL-10 gene treatment subgroup compared with the fibrosis model and empty vector control subgroup .Immunohistochemistry analysis showed that the expression of MMP 13 and TIMP1 in fibrosis model subgroup and empty vector control subgroup was significantly higher than the normal control group , but compared with normal control group , expression of MMP13 was significantly increased and expres-sion of TIMP1 was significantly decreased in rIL-10 gene treatment subgroup .Compared with fibrosis model subgroup and empty vector control subgroup, the expression of MMP13 and TIMP1 was dramatically decreased in rIL-10 gene treatment subgroup.Conclusion:rIL-10 gene treatment attenuates the area of collagen deposition in liver fibrosis rats associated with downregulation of TIMP 1.

Effects of small interfering RNA targeting hepatitis B virus X protein on mitochondrial function in healthy liver cell line steadily expressed X gene / 中华传染病杂志

Objective To construct and identify recombinant expression plasmid of small interfering RNA (siRNA)targeting hepatitis B virus X protein(HBx), and observe its effect on mitoehondrial function in healthy liver cell line steadily expressed HBx gene (HL-7702/HBx). Methods Two siRNA sequences containing short hairpin structure, which target on the total length HBx gene, were synthesized and cloned into the vector psiRNA-Hh1GFPzeo to eonstruct recombinant expression plasmids pX1 and pX2. Non-specific recombinant pScr plasmid served as control. After siRNA transfected into HL-7702/HBx cells line by liposome, reverse transcription-polymerase chain reaction (RT-PCR) and Western blot were performed to identify the suppressive effect on HBx expression. Levels of intraeellular reactive oxygen species (ROS) and mitochondrial membrane potential (△ m) were determined by flow cytometry. The experimental results were compared by analysis of variance. Results Successful constructions of pX1 and pX2 were confirmed by restriction enzyme digestion and sequencing. The expressions of HBx mRNA and protein after 48 h of transfection into HL-7702/HBx cells in control group were 0.65± 0.12 and 0.62± 0.09, respectively, which were both higher than those (0.33±0.10 and 0.19±0.08, respectively) in group pX1 (t=4.73, P<0.05; t=7.53, P<0.05) and those (0.48±0.10 and 0.37±0.11, respectively) in group pX2 (t=2.39, P<0.05;t=4.43,P<0.05). But the inhibition of group pX1 was stronger than that of pX2 (t=2.28,P<0.05). Levels of ROS and △ m after RNA interference were 5.00±0.38 and 33.86±0.50, respectively, while those in control group were 72. 10±0. 55 and 3. 57±0.26, respectively (ROS: t=276.22, P<0.05; △ m: t=107.15, P<0.05). Conclusions siRNA targeting HBx can efficiently and specifically suppress the HBx expression in HL-7702/HBx cells, and decrease the level of ROS and increase the level of △ m, thus relieve cellular oxidative stress.

EXPERIMENTAL STUDY ON THE RELATIONSHIP OF TWO DIFFERENT CHOLECYSTOJEJUNOSTOMIES AND BILIARY RETROGRADE INFECTION / 现代医院

Objective To compare the prevent effect of biliary retrograde infection in the uncut and cut Roux-en-Y cholecystojejunostomy. Methods Sixteen Begle dogs were divided into two groups randomly. One group was performed with the uncut Roux-en-Y cholecystojejunostomy(the uncut group) and the other was performed with the cut Roux-en-Y cholecystojejunostomy (the cut group). The categories of bacterium in the the Roux limb before and twelve weeks after operation, and the biliary duct pressure of the anastomotic stoma were detected. The changes of the categories of bacterium before and after operation, the categories of bacterium and the biliary duct pressure after operation in two groups were analyzed. Results The isolating rate of G-aerobic bacteria and anaerobic bacteria was 37.50% and 6.25% before operation; in the uncut group, the isolating rate was 62.50%, 12.50%,with no significance difference comparing to preoperation(p＝0.390、1.000). In the uncut group the isolating rate was 87.50%, 75.00%after operation, with significance difference comparing to preoperation (p＝0.033、0.001).The isolating rate of anaerobic bacteria after operation in the cut group is higher than the uncut group,the difference is significance (p＝0.041). And the biliary duct pressure in the cut group is higher than the uncut group(p<0.001).Conclusion The uncut Roux-en-Y cholecystojejunostomy was more effective in prevent biliary retrograde infection.

The influence of the tyrosine Kinase inhibitor imatinib mesylate on the cell repair function after DNA damage in K562 cells / 中华检验医学杂志

Objeetive To construct the cell DNA damage models for the human CML K562 cell line before or after imarlnib mesylate treatment and observe the repairing process dynamically for investigating the iniluence of imatinib mesylate on the repair function of K562 cells after cell DNA danlage.Methods The MTT assay was used to estimate the optimal pretreatment concentration of imatinib mesylate in K562 cells and Western blot was employed to evaluate the phosphorylation status in K562 cells after imatinib mesylate treatment to estimate BCR/ABL tyrosine kinase inhibition by imatinib mesylate.The comet assay was used to detect the DNA damage induced by hydrogen peroxide at various concentrations in K562 cells with or without the pretreatment of imatinib mesylate.A dynamic observation on the repairing process after cell DNA damage was made by the comet assay.Results The pretreatment by imatinib mesylate for K562 cells was optimized to be at a final concentration of 1 μmol/L for 24 h as revealed by the MTT assay additionly imatinib mesylate treatment at this concentration could effectively inhibit the phosphorylation of the BCR-ABL fusion protein at Tyr177(Deusityrate 0.100±0.018).When compared with the control group(Deusityrate 0.425±0.039),the BCR/ABL phosphorylation at Tyr177 was significantly decreased by (77. 11±5.59) % (t=4. 57,P＜0. 05). The cell DNA damage models for both imatinib mesylate-nontreated and imatinib mesylate-pretreated K562 cell groups were constructed with hydrogen peroxide treatment at a final concentration of 10 μmol/L for 10 min at 4℃ as confirmed by the comet assay. When compared with the control imatinib mesylatenontreatod K562 cell group,the time duration required for the DNA repair in imatinib mesylate-pretreated K562 cell group was significantly prolonged (F= 97.79,P＜0. 05 ). Conclusions The cell DNA damage models for the leukemic K562 cell groups before or after imatinib mesylate treatment were successfully constructed and the tyrosine kinase inhibitor imatinib mesylate for BCR/ABL fusion protein was revealed to attenuate the DNA repair capacity of the K562 cells after DNA damage.