[Application of acute physiology and chronic health evaluation II score in the timing of noninvasive ventilation in patients with acute exacerbation of chronic obstructive pulmonary disease].

OBJECTIVE: To explore the application of acute physiology and chronic health evaluation II (APACHE II) score in the timing and nursing of noninvasive ventilation for patients with acute exacerbation of chronic obstructive pulmonary disease (AECOPD). METHODS: 106 AECOPD patients admitted to Haikou People's Hospital from January 2018 to October 2019 were selected as the study objects. According to the method of random number table, the patients were divided into observation group and control group, with 53 patients in each group. The control group selected the timing of noninvasive ventilation treatment according to the standards of Mechanical ventilation (second edition), weaned according to Clinical practice of mechanical ventilation, and received routine nursing in intensive care unit (ICU), including creating comfortable indoor environment, reasonable diet, condition monitoring, psychological nursing and complications nursing. On the basis of the control group, the patients in the observation group were given noninvasive ventilation when APACHE II score was more than 10, and were weaned when APACHE II score was less than or equal to 10. According to APACHE II score < 10, 10-14, 15-19 and ≥ 20, the patients were given level-3 care, level-2 care, level-1 care and intensive care. The pulmonary function before and 3 days after the noninvasive ventilation treatment was monitored, and the duration of mechanical ventilation, the length of ICU stay, endotracheal intubation rate, incidence of complication [ventilator associated pneumonia (VAP)] and ICU mortality were recorded. The self-designed questionnaire of nursing satisfaction was used to evaluate the patients' nursing satisfaction. RESULTS: There was no significant difference in general data such as gender or age between the two groups. After 3 days of noninvasive ventilation, the forced expiratory volume in one second (FEV1), forced vital capacity (FVC) and FEV1/FVC ratio of the two groups were increased significantly as compared with those before treatment, especially in the observation group, with statistical significances as compared with the control group [FEV1 (L): 3.02±0.22 vs. 2.54±0.19, FVC (L): 3.01±0.32 vs. 2.13±0.28, FEV1/FVC ratio: 0.89±0.08 vs. 0.79±0.08, all P < 0.05]. Compared with the control group, the duration of mechanical ventilation and length of ICU stay in the observation group were significantly shortened [duration of mechanical ventilation (days): 4.32±0.73 vs. 8.42±1.94, length of ICU stay (hours): 32.23±10.22 vs. 38.52±9.85, both P < 0.01]. The intubation rate, incidence of VAP and ICU mortality in the observation group were significantly lower than those in the control group [intubation rate: 1.9% (1/53) vs. 13.2% (7/53), incidence of VAP: 1.9% (1/53) vs. 15.1% (8/53), ICU mortality: 1.9% (1/53) vs. 13.2% (7/53), all P < 0.05]. The nursing satisfaction of patients in the observation group was significantly higher than that in the control group [96.2% (51/53) vs. 75.5% (40/53), P < 0.01]. CONCLUSIONS: APACHE II score can be used to guide the choice of noninvasive ventilation treatment opportunity and nursing intervention measures for AECOPD patients. It can significantly improve the pulmonary function of patients, improve the treatment effect, reduce the incidence of complications, and improve the satisfaction of patients with nursing, which is effective in clinical application.

[Etiological analysis of 1977 patients with acute liver failure, subacute liver failure and acute-on-chronic liver failure].

OBJECTIVE: To investigate the etiology of 1977 patients from northern China with acute (ALF), sub-acute (SALF) or acute-on-chronic liver (ACLF) failures. METHOD: The age, gender, etiology, pathogenesis, and prognosis of the 1977 patients with liver failures were retrospectively analyzed. RESULTS: Of the 1977 cases, the three most common causes of ALF were HEV (33.96%) or HBV (13.21%) infections or those caused by medicines (9.43%). The three predominant causes of SALF were medicines (31.53%), HEV (16.22%) or HBV (9.91%) infections, but those of the ACLF were HBV (90.29%) infection, alcoholic hepatopathy (2.65%), and HBV super infected with HEV (2.26%) infections. 90.09% (1781) patients were infected by hepatotropic viruses. Of these 1781 patients, the most common cause of their liver failures was HBV infection (92.93%). In these HBV infected patients, 77.10% were from 26 to 55 years old. From 2005 to 2007, there were 39 patients with alcoholic liver failure. In the past two years, there were 23 patients with drug induced liver failure. The improvement rate of the 1977 patients after their treatments was 35.56%. The improvement rate of HEV infected liver failure was higher than drug induced liver failure (P less than 0.05); no statistical significance was found between other groups (P more than 0.05). CONCLUSION: Different types of liver failure have different predominant causes. HBV infection is the most common cause in our 1977 patients. In the past two years, the number of drug induced liver failures and alcoholic liver failures have been increasing.

Prognostic factors for severe acute respiratory syndrome: a clinical analysis of 165 cases.

This study analyzes single factors that affect the prognosis of severe acute respiratory syndrome (SARS) and establishes a prognosis model by multivariate analysis. We retrospectively analyzed the clinical features of SARS in 165 clinically confirmed severe cases. Both age and existence of other diseases before SARS were significantly correlated with prognosis (r=0.506 and r=0.457, respectively; P<.001). During the acute phase of SARS, lactate dehydrogenase level, degree of hypoxemia, respiratory rate, alpha -hydroxybutyric dehydrogenase level, creatine kinase isoenzyme-MB, platelet count, and number of involved lobes noted on chest radiographs, and so on, correlated markedly with the prognosis (r=0.257-0.788; P<.05). The multivariate prognosis regression model was associated with degree of hypoxemia and platelet count. The model was defined by the formula Py=1=es/(1+es), where S is [2.490 x degree of hypoxemia]-[0.050 x number of platelets], and it had a high sensitivity (91.67%), specificity (98.33%), and accuracy (96.42%). The model could be used to effectively judge the state of illness and the prognosis.

Preparation of human single chain Fv antibody against hepatitis C virus E2 protein and its identification in immunohistochemistry.

AIM: To screen human single chain Fv antibody (scFv) against hepatitis C virus E2 antigen and identify its application in immunohistochemistry. METHODS: The phage antibody library was panned by HCV E2 antigen, which was coated in microtiter plate. After five rounds of biopanning,56 phage clones were identified specific to HCV E2 antigen. The selected scFv clones were digested by SfiI/NotI and DNA was sequenced. Then it was subcloned into the vector pCANTAB5E for expression as E-tagged soluble scFv. The liver tissue sections from normal person and patients with chronic hepatitis B and chronic hepatitis C were immunostained with HCV E2 scFv antibody. RESULTS: The data of scFv-E2 DNA digestion and DNA sequencing showed that the scFv gene is composed of 750 bp. ELISA and immunohistochemistry demonstrated that the human single chain Fv antibody against hepatitis C E2 antigen has a specific binding character with hepatitis virus E2 antigen and paraffin-embedded tissue, but did not react with liver tissues from healthy persons or patients with chronic hepatitis B. CONCLUSION: We have successfully screened and identified HCV E2 scFv and the scFv could be used in the immunostaining of liver tissue sections from patients with chronic hepatitis C.

Antitumor activities of human autologous cytokine-induced killer (CIK) cells against hepatocellular carcinoma cells in vitro and in vivo.

AIM: To characterize the anticancer function of cytokine-induced killer cells (CIK) and develop an adoptive immunotherapy for the patients with primary hepatocellular carcinoma (HCC), we evaluated the proliferation rate, phenotype and the antitumor activity of human CIK cells from healthy donors and HCC patients in vitro and in vivo. METHODS: Peripheral blood mononuclear cells (PBMC) from healthy donors and patients with primary HCC were incubated in vitro and induced into CIK cells in the presence of various cytokines such as interferon-gamma (IFN-gamma), interleukin-1 (IL-1), IL-2 and monoclonal antibody (mAb) against CD3. The phenotype and characterization of CIK cells were identified by flow cytometric analysis. The cytotoxicity of CIK cells was determined by (51)Cr release assay. RESULTS: The CIK cells were shown to be a heterogeneous population with different cellular phenotypes. The percentage of CD3+/CD56+ positive cells, the dominant effector cells, in total CIK cells from healthy donors and HCC patients, significantly increased from 0.1-0.13% at day 0 to 19.0-20.5% at day 21 incubation, which suggested that the CD3+ CD56+ positive cells proliferated faster than other cell populations of CIK cells in the protocol used in this study. After 28 day in vitro incubation, the CIK cells from patients with HCC and healthy donors increased by more than 300-fold and 500-fold in proliferation cell number, respectively. CIK cells originated from HCC patients possessed a higher in vitro antitumor cytotoxic activity on autologous HCC cells than the autologous lymphokine-activated killer (LAK) cells and PBMC cells. In in vivo animal experiment, CIK cells had stronger effects on the inhibition of tumor growth in Balb/c nude mice bearing BEL-7402-producing tumor than LAK cells (mean inhibitory rate, 84.7% vs 52.8%, P<0.05) or PBMC (mean inhibitory rate, 84.7% vs 37.1%, P<0.01). CONCLUSION: Autologous CIK cells are of highly efficient cytotoxic effector cells against primary hepatocellular carcinoma cells and might serve as an alternative adoptive therapeutic strategy for HCC patients.

Interaction between hepatitis C virus core protein and translin protein--a possible molecular mechanism for hepatocellular carcinoma and lymphoma caused by hepatitis C virus.

AIM: To investigate the interaction between hepatitis C virus core protein and translin protein and its role in the pathogenensis of hepatocellular carcinoma and lymphoma. METHODS: With the components of the yeast two hybrid system 3, "bait" plasmids of HCV core the gene was constructed. After proving that hepatitis C virus core protein could be firmly expressed in AH109 yeast strains, yeast two- hybrid screening was performed by mating AH109 with Y187 that transformed with liver cDNA library plasmids-pACT2 and then plated on quadruple dropout (QDO) medium and then assayed for alpha-gal activity. Sequencing analysis of the genes of library plasmids in yeast colonies that could grow on QDO with alpha-gal activity was performed. The interaction between HCV core protein and the protein we obtained from positive colony was further confirmed by repeating yeast two - hybrid analysis and coimmunoprecipitation in vitro. RESULTS: A gene from a positive colony was the gene of translin, a recombination hotspot binding protein. The interaction between HCV core protein and translin protein could be proved not only in yeast, but also in vitro. CONCLUSION: The core protein of HCV can interact with translin protein. This can partly explain the molecular mechanism for hepatocellular carcinoma and lymphoma caused by HCV.

Pathological changes and clinical manifestations of 1020 children with liver diseases confirmed by biopsy.

BACKGROUND: Liver biopsy plays an important role in accurate diagnosis of various liver diseases in children and liver damages caused by systemic illnesses. This study was designed to evaluate the value of liver biopsy in diagnosis of liver diseases in children and explore the relationship between their pathological changes and clinical manifestations. METHODS: One-second liver biopsy was performed in 1023 pediatric patients with liver diseases at our department from 1983 to 2000. Diagnosis of viral hepatitis was based on the diagnostic criteria formulated by the Chinese Society of Infectious and Parasitic Diseases in 1995. Inflammatory changes of the liver were graded from 0 to 4 (G0-4). RESULTS: Liver biopsy was performed successfully in 1020 patients including 135 infants and young children, of whom 90% were hospitalized patients with chronic liver diseases. Hepatitis virus was the leading cause for chronic liver diseases, among which hepatitis B was detected in 75.4% of the patients. Sixty-nine patients showed liver impairment induced by disorders relevant to that metabolism, Wilson's disease, and glycogen storage disease. Liver inflammatory injury (

Screening of augmenter of liver regeneration-binding proteins by yeast-two hybrid technique.

OBJECTIVE: To investigate the biological function of augmenter of liver regeneration (ALR), we used yeast-two hybrid technique to detect proteins in hepatocytes interacting with ALR. METHODS: ALR bait plasmid was constructed by using yeast-two hybrid system 3, then transformed into yeast AH109. The transformed yeast was mated with yeast Y187 containing liver cDNA library plasmid in a 2XYPDA medium. Diploid yeast was plated on a synthetic dropout nutrient medium (SD/-Trp-Leu-His-Ade) containing x-alpha-gal for selection and screening. After extracting and sequencing of the plasmid from blue colonies. Analysis was performed by bioinformatics. RESULTS: Of 36 colonies sequenced, 14 are metallothionein, 12 albumin, and 3 selenoprotein P. One colony is a new gene with unknown function. CONCLUSION: The successful cloning of gene of ALR interacting protein has paved the way for studying the physiological function of ALR and associated proteins.

[Characterization of peripheral dendritic cell subsets and its implication in patients infected with severe acute respiratory syndrome].

OBJECTIVE: To investigate the dynamic changes of dendritic cell subsets in peripheral blood of patients infected with severe acute respiratory syndrome (SARS) and evaluate their roles in the immunopathogenesis of SARS. METHODS: Flow cytometry was applied to study the dynamic alteration of the number and frequencies in circulating DC cell subsets in 30 SARS patients including critical SARS (n = 11) and general SARS (n = 19). The reasons and clinic significances of the peripheral blood DC subsets changes in SARS patients were also analyzed in our study. RESULTS: The patients in critical status had a 9-week course of disease, longer than the 6-week course observed in subjects in general status. The frequency of peripheral DC cell subsets significantly dropped beginning from the onset of symptom in SARS patients and was maintained at significant low levels during the following 4 - 5 weeks, 1.7 +/- 1.8, 5.3 +/- 5.0/ micro l for DC1, 0.57 +/- 1.02, 0.98 +/- 1.11/ micro l for DC2 for cases in critical and general statuses, respectively, compared with healthy subjects; more importantly, the pDC2 even disappeared in the patients who died from SARS diseases. The possible reasons responsible for the alteration of DC subsets in peripheral blood is likely to be the direct attack of SARS-CoVin circulation and be partially involved the application of large dose of steroid. The frequency in DC cell subsets returned to normal level in convalescent stage. CONCLUSION: Our results showed SARS patients had a significant decrease of circulating DC cell subset frequency, which maybe lead to the host immunodeficiency response to SARS-associated coronavirus (SARS-CoV).

[Mutual interaction between hepatitis C virus core protein and translin, a recombination hotspot binding protein].

OBJECTIVE: To study the role of the core protein of hepatitis C virus (HCV) in HCV pathogenesis and investigate the molecular mechanism of this protein in tumorigenesis. METHODS: By using the yeast two hybrid system 3, the bait plasmids of the gene of the core protein of HCV was constructed and transfected into AH109 yeast strain. Then this transfected AH109 yeast was mated with Y187 yeast containing liver cDNA library plasmids and cultured on quadropledropout (QDO) medium covered with x-alpha-gal and assayed for alpha-gal activity. The blue colonies growing on QOD were collected to extract the plasmids to transform Escherichia coli. Plasmids were extracted from the bacterium and sequenced. After bioinformatic analysis translin, a recombination hotspot binding protein interacting with HCV core protein was found. To further prove the interaction between translin protein and HCV core protein translation was performed by using reticulocyte lysate and immunoprecipitation in vitro. RESULTS: Among the 30 positive colonies, a colony was translin. The interaction between HCV core protein and translin protein could be proved not only in yeast, but also in vitro. CONCLUSION: The core protein of HCV can interact with translin protein, this can explain partly the tumorigenesis of HCV.

[Preparation of human single chain Fv antibody specifically against hepatitis C virus E2 antigen].

OBJECTIVE: To identify human single chain Fv antibody (ScFv) against hepatitis C viral E2 antigen and its value clinically. METHODS: The recombinant phages were panned by E2 antigen which was coated in a microtiter plate. After five rounds of biopanning, 56 phage clones were identified specific to E2 antigen. The affinity and specificity of ScFv were evaluated by ELISA and immunohistochemistry, respectively. RESULTS: The data of E2-ScFv DNA digestion and DNA sequencing showed that the ScFv gene was composed of 750bp. ELISA and immunohistochemistry demonstrated that the human single chain Fv antibody against HCV E2 antigen had a specific combination character with hepatitis C virus E2 antigen. CONCLUSIONS: ScFv, having a sutestantial affinity and specificity and being easy to prepare, is valuable in the detection of HCV E2 antigen.

[Cloning of genes transactivated by hepatitis B virus X protein].

OBJECTIVE: To construct a subtractive cDNA library of genes transactivated by hepatitis B virus X protein (HBX) using suppression subtractive hybridization (SSH) technique and to clone genes associated with HBX transactivating function. METHODS: The mRNA was isolated from HepG2 cells transfected with pcDNA3.1(-)-X and pcDNA3.1(-) empty vector respectively, then cDNA was synthesized. After restriction enzyme RsaI digestion, a number of small size cDNA was obtained. Then tester cDNA was subdivided into two portions and each was ligated with different cDNA adaptor. After tester cDNA was hybridized with driver cDNA twice and underwent nested polymerase chain reaction (PCR) twice the production was subcloned into T/A plasmid vectors to set up the subtractive cDNA library. Amplification of the library was carried out with E. coli strain JM109, some cDNA was sequenced and analyzed in GenBank with Blast. RESULTS: The subtractive cDNA library of genes transactivated by HBX was constructed. The amplified library contained 85 positive clones, and colony PCR showed that these clones contained 200-1000 bp inserts. 65 clones were analyzed by sequencing and bioinformatics, which suggested nineteen known genes and fifteen genes with unknown function. CONCLUSION: A subtractive cDNA library of genes transactivated by HBX using SSH technique has been constructed successfully, which may bring some new clues for studying the biological functions of HBX and the pathogenesis of hepatoma.

[Synergetic transactivating functions of HCV core and truncated HBV middle surface protein].

OBJECTIVE: To investigate the synergetic transactivating functions of HCV core and truncated HBV middle surface proteins. METHODS: Two recombinant expression plasmids harboring HCV core and C-terminally truncated HBV middle surface protein gene were constructed, respectively. The plasmids were transfected into HepG2 cells and cotransfected HepG2 cells with reporter plasmid pSV-lacZ by lipofectamine plus reagents. The transient expressed viral proteins were identified at the transcription and translation levels. The activity of beta-galactosidase was detected, which reflected the transactivating function of the proteins. RESULTS: The protein expression of plasmids was detected in soluble cell extracts of transiently transfected HepG2 cells. HCV core protein activated the beta-galactosidase expression at a value of 4.6 times higher than the control, while C-terminally truncated HBV middle surface protein activated at a value of 3.2 times. It reached 8.4 times transfected with the plasmids simultaneously. The transactivating effect was dose dependent. CONCLUSIONS: It is suggested that the two kinds of virus proteins have transactivating effect on SV40 early promoter/enhancer, and they act synergistically. These contribute to explain the mechanisms of liver injury or tumorigenesis induced by HCV or/and HBV infection.

[Lamivudine and entecavir significantly improved the prognosis of early-to-mid stage hepatitis B related acute on chronic liver failure].

OBJECTIVE: To clinically study the antiviral effects of lamivudine and entecavir on patients with early-to-mid stage Hepatitis B related acute on chronic liver failure (HBV-ACLF). METHODS; A prospective, randomized, open and parallel controlled clinical trial was designed to observe the antiviral effects of nucleoside analogues on patients with early-to-mid stage HBV-ACLF. Three groups were set for controlled study, i. e. basic treatment group, lamivudine plus basic treatment group and entecavir plus basic treatment group. RESULTS: One month after treatment, the improvement rates of lamivudine group and entecavir group were 58.85% and 59.15% respectively, significantly higher than that of basic treatment group which was 34.84% (Chi(2) = 9.8323, P = 0.043). By the end of six months, the cumulative survival rates of patients with the antiviral treatments, i.e., lamivudine, entecavir, were 65.8%, 60.1%, significantly higher than that (42%) without the antiviral treatment (P = 0.045, P = 0.04 respectively). The cumulative survival rate in patients with a MELD score < 30 was higher than that with a MELD score over 30 (Chi(2) = 3.920, P = 0.048). For the patients with pretreatment HBV DNA > or = 10(7), the cumulative survival rate in patients with entecavir treatments group was higher than that of patients in basic treatment group (Chi(2) = 5. 014 P= 0.025). According to the Ordinal Regression analysis, antiviral therapy by using either lamivudine or entecavia could significantly increase the improvement rate of patients with early-to-mid stage HBV-ACLF. But severe complications, including hepatorenal syndrome, electrolyte imbalance and hepatic encephalopathy, medical history of liver cirrhosis, and pretreatment HBV DNA > or = 10(7) had significant impacts on prognosis of this group patients. CONCLUSIONS: Antiviral therapy by using either lamivudine or entecavia could significantly increase the survival rate of patients with early-to-mid stage HBV-ACLF.

[Relation of viral genotypes to clinical features in children with chronic hepatitis B].

OBJECTIVE: To investigate the relation of HBV genotypes to clincal features in children with chronic hepatitis B. METHODS: The genotypes of serum HBV DNA from 404 children with chronic hepatitis B were determined by PCR using type-specific primers. RESULTS: For the 404 children, genotype B in 99 (24.5%), genotype C in 285 (70.5%). For the 75 children from south part of China, 29 were of genotype B (38.7%) and 44 of genotype C (58.7%). For the 329 children from north part of China, 70 were of genotype B (21.3%) and 241 of genotype C (73.3%). There were significant differences between the children from south part and those from north part of China in genotype B and C (P = 0.002). Genotype B and C were not significantly correlated to gender, age and mother-to-fetus transmission. There was no marked difference in liver injury severity (P = 0.4796), serum HBeAg positivity, HBVDNA level, inflammatory degree of liver tissue (P = 0.209) and liver fibrosis( P = 0.177) between the children with genotype B and those with genotype C. CONCLUSION: In children with HBV infection, genotype C accounts for 70.5% and genotype B for 24.5%. The genotypes are of regional difference in children with HBV infeciton. There are replication and liver pathological change between genotype B and genotype C.

[To re-evaluate the clinical classification criteria of liver failure].

OBJECTIVE: To study the clinical feature and more reasonable diagnostic typing criteria for patients with liver failure. METHODS: 13/21 cases of ALF, SALF with no past liver disease, 49/72 cases of with chronic hepatitis, and 23/73 cases ALF, SALF with liver cirrhosis, were analyzed respectively. RESULTS: 1 ALF patients (1). There exist significant statistic differences in ALB, ALT, CHE in three ALF groups.(2). It had statistic differences in those patients with hepatic encephalopathy.(3). The prognosis of the patients with chronic hepatitis group (42.85 percent) was best than that of chronic cirrhosis (26.09 percent) and no past liver disease (15.38 percent). (2) In SALF patients (1). There exist significant statistic differences in ALB, GLO, ALT, AST, BDIL, GLU and CHE in three SALF groups.(2). It had statistic differences in those patients with hepatic encephalopathy in three SALF groups.(3). The prognosis of the patients with chronic hepatitis group (51.39 percent) was best than that of chronic cirrhosis (36.85 percent) and no past liver disease (33.33 percent). CONCLUSION: There are different clinic feature and prognosis in three ALF or SALF groups, so we suggest that it were clinic practicability and science in classify of liver failure at present.

[Efficacy and safety of Fufang Biejia Ruangan tablet in patients with chronic hepatitis B complicated with hepatic fibrosis].

OBJECTIVE: To study the clinical therapeutic effects and safety of Fufang Biejia Ruangan tablet (FBRt) in patients with chronic hepatitis B complicated with hepatic fibrosis. METHODS: Totally 420 patients were randomly divided into two groups, FBRt group (300 cases) were treated with Fufang Biejia Ruangan tablets and control group (120 cases) were treated with He Luo Shu Gan capsule, the patients in both groups were treated for 6 months. RESULTS: The cure rate and total effective rate of FBRt group were significantly higher than those of control group (55.67 percent and 81.67 percent vs. 15.8 percent and 60.00 percent, P less than 0.01). CONCLUSION: Fufang Biejia Ruangan tablet could alleviate clinical symptoms and hepatic fibrosis. Fufang Biejia Ruangan tablet is effective and safe in treatment of patients with chronic hepatitis B complicated with liver fibrosis.