Real-world tobacco cessation practice in China: findings from the prospective, nationwide multicenter China National Tobacco Cessation Cohort Study (CNTCCS).

Background: Tobacco cessation is proven to be the most effective and cost-effective strategy for smokers to reduce their risk of smoking-related disease and premature death. Providing effective, efficient, safe, and patient-centred tobacco cessation treatment to reach those who need them is a significant challenge. To date, only a few nationwide studies in China have assessed the overall clinical care practice and treatment outcome of tobacco cessation. Methods: This a prospective, nationwide, multicenter, cohort study covering all Eastern China, Northwest China, Central China, North China, Southwest China, Northeast China, and South China. Participants who were current smokers aged 18-85 years attending clinic for smoking cessation were included. All the participants were treated with 3-month cessation treatment and followed up for 3 months. Data were collected prospectively using online system. The primary outcome was 7-day point abstinence rate at 24 weeks, validated biochemically by an expired carbon monoxide level of less than 10 ppm. The participants lost to follow-up or not providing validation were included as non-abstainers. Findings: A representative sample of 3557 participants were recruited and 2943 participants were included into this analysis. These participants had mean age of 53.05 years, and 94.8% were males, with 75.8% showing symptoms of tobacco dependence. A total of 965 (32.8%) participants were treated with Bupropion + behavioural counselling, followed by 935 (31.8%) with behavioural counselling, 778 (26.4%) with Varenicline + behavioural counselling, 135 (4.6%) with alternative treatments + behavioural counselling, and 130 (4.4%) with nicotine replacement therapy (NRT) + behavioural counselling. After 3-month treatment and 3-month follow-up, 21.74% of the participants quit smoking at 24 weeks. In the multivariable-adjusted analyses, quitting smoking was significantly associated with female, higher socioeconomic status, poor health condition, different treatment received, and less smoking intensity. The tobacco cessation treatment varied widely across different areas of China. In particular, the areas with higher usage of cessation medication were associated with better cessation treatment outcome. Interpretation: The CNTCCS is the first large-scale nationwide cohort study of smoking cessation in China. Rich data collected from this prospective cohort study provided the opportunity to evaluate the clinical practice of tobacco cessation treatment in China. Funding: Chinese Academy of Medical Sciences (CAMS) Initiative for Innovative Medicine (CAMS 2021-I2M-1-010), Heilongjiang Provincial Science and Technology Key Program (2022ZXJ03C02), and National Key R&D Program of China (grant no. 2017YFC1309400).

PTH Predicts the in-Hospital MACE After Primary Percutaneous Coronary Intervention for Acute ST-Segment Elevation Myocardial Infarction.

Objective: To investigate the correlation between serum parathyroid hormone (PTH) levels and in-hospital major adverse cardiovascular events (MACE) in patients with acute ST-segment elevation myocardial infarction (STEMI) after primary percutaneous coronary intervention (PCI), and establish a risk prediction model based on parameters such as PTH for in-hospital MACE. Methods: This observational retrospective study consecutively enrolled 340 patients who underwent primary PCI for STEMI between January 2016 and December 2020, divided into a MACE group (n=92) and a control group (n=248). The least absolute shrinkage and selection operator (LASSO) and logistic regression analyses were used to determine the risk factors for MACE after primary PCI. The rms package in R-studio statistical software was used to construct a nomogram, to detect the line chart C-index, and to draw a calibration curve. The decision curve analysis (DCA) method was used to evaluate the clinical application value and net benefit. Results: Correlation analysis revealed that PTH level positively correlated with the occurrence of in-hospital MACE. Receiver operating characteristic curve analyses revealed that PTH had a good predictive value for in-hospital MACE. Multivariate logistic regression analysis indicated that Killip class II-IV, and FBG were independently associated with in-hospital MACE after primary PCI. A nomogram model was constructed using the above parameters. The model C-index was 0.894 and the calibration curve indicated that the model was well calibrated. The DCA curve suggested that the nomogram model was better than TIMI score model in terms of net clinical benefit. Conclusion: Serum PTH levels in patients with STEMI are associated with in-hospital MACE after primary PCI, and the nomogram risk prediction model based on PTH demonstrated good predictive ability with obvious clinical practical value.

Application of short hydrophobic elastin-like polypeptides for expression and purification of active proteins.

To investigate the application of short elastin-like polypeptides (ELPs) in the purification of bioactive proteins, short hydrophobic ELP[I] n (n = 30, 40, 50) tags were constructed. Both the ELP[I] n tags and the ELP[I] n -Trx fusion proteins could be stably expressed in Escherichia coli and purified by inverse transition cycling, respectively. Total protein concentrations determined by BCA protein assay showed that the yield of the fusion proteins decreased with increasing ELP length. Measurements of the inverse transition temperature (T t) of the ELP[I] n -Trx under different salts or PEG8000 concentrations showed decreased T t upon elevated concentrations; while, all the T ts were suitable for generating proteins from 4 to 37.5 ºC. Furthermore, to identify a linker peptide for bioactive protein production without the need to remove the ELP[I] n tag, the activity of eGFP protein fused with ELP[I]30 tag by either a poly-N or a G4S linker was quantified using a fluorescence spectrophotometer. The results indicated that the ELP[I]30-eGFP fusion proteins with the poly-N linker showed higher fluorescence levels than those with the G4S linker. Our results demonstrated that short ELP[I] n tags with low T t were useful in protein expression and purification, and poly-N linker played the key role in producing bioactive proteins without the need to remove the ELPs.

TRIM22 confers poor prognosis and promotes epithelial-mesenchymal transition through regulation of AKT/GSK3ß/ß-catenin signaling in non-small cell lung cancer.

Expression pattern and biological roles of TRIM22 remains unknown in most human cancers. The present study aims to discover its clinical significance and function in human non-small cell lung cancer (NSCLC). Immunohistochemistry was used to examine TRIM22 expression in 126 cases of NSCLC specimens. TRIM22 protein was upregulated in 70/126 (55.6%) non-small cell lung cancer tissues compared with normal lung tissue. TRIM22 overexpression was associated with advanced TNM stage, positive nodal metastasis and poor prognosis. Plasmid and siRNA transfection were performed in lung cancer cell lines. TRIM22 overexpression promoted proliferation, colony formation and invasion in A549 cells. While its depletion exhibited the opposite effects in H1299 cell line. TRIM22 overexpression promoted cell cycle progression through regulation of cyclin D1, cyclin E and p27. TRIM22 also changed the expression of epithelial to mesenchymal transition (EMT) markers including E-cadherin N-cadherin, Vimentin and Snail. Furthermore, TRIM22 activated PI3K/AKT/GSK3ß/ß-catenin oncogenic signaling pathways. Treatment with PI3K inhibitor LY294002 and ß-catenin siRNA blocked the effects of TRIM22 on EMT in TRIM22-overexpressing cells. In conclusion,TRIM22 serves as an important oncoprotein and a promoter of cell proliferation and invasion through AKT/ GSK3ß/ß-catenin induced EMT in NSCLC.

Analysis of hospitalization expenditures and influencing factors for inpatients with coronary heart disease in a tier-3 hospital in Xi'an, China: A retrospective study.

BACKGROUND: The medical costs for inpatients with coronary heart disease (CHD) have risen to unprecedented levels, putting tremendous financial pressure on their families and the entire society. The objective of this study was to examine the actual direct medical costs of inpatients with CHD and to analyze the influencing factors of those costs, to provide advice on the prevention and control of high medical costs of patients with CHD. METHODS: A retrospective descriptive analysis of hospitalization expenditures data examined 10,301 inpatients with coronary heart disease of a tier-3 hospital in Xi'an from January 1, 2015 to December 31, 2015. The data included demographic information, the average length of stay, and different types of expenses incurred during the hospitalization period. The difference between different groups was analyzed using a univariate analysis, and the influencing factors of hospitalization expenditures were explored by the multiple linear stepwise regression analysis. RESULTS: The average age of these patients was 60.0 years old, the average length of stay was 4.0 days, and the majority were males (7172, 69.6%). The average hospitalization expenses were $6791.38 (3294.16-9, 732.59), and the top 3 expenses were medical consumables, operation fees, and drugs. The influencing factors of hospitalization expenditures included the length of stay, the number of times of admission, the type of medical insurance schemes, whether have a surgery or not, the gender, the age, and the marriage status. CONCLUSION: The inpatients with CHD in this tier-3 hospital were mostly over 45 years old. The average medical cost of males was much higher than that of females. Our findings suggest that the solution for tremendous hospitalization expenditures should be that more attention is paid to controlling the high expense of medical consumables and that the traditional method of reducing medical expenses by shortening the length of stay is still important in nowadays. Furthermore, the type of medical insurance schemes has different impacts on medical expenses. Reducing or controlling high hospitalization expenditures is a complicated process that needs multifaceted cooperation.

The optimal sequence for bronchial brushing and forceps biopsy in lung cancer diagnosis: a random control study.

BACKGROUND: Optimizing basic techniques in diagnostic bronchoscopy is important for improving medical services in developing countries. In this study, the optimal sequence of bronchial brushing relative to bronchial biopsy for lung cancer diagnosis was evaluated. METHODS: A total of 420 patients with visible endobronchial tumors were prospectively and randomly enrolled in two groups: a pre-biopsy brushing group, receiving two brushings before biopsy; two brushings which performed afterwards; were set as self-control and compared with the pre-biopsy brushings as the intra-group comparison; and a post-biopsy brushing group, only receiving two brushings after biopsy, which were compared with the pre-biopsy brushings as the inter-group comparison. Diagnostic yield of brushing was compared before and after biopsy, and as well as for different tumor pathologies and bronchoscopic morphologies. The occurrence of treated bleeding which defined as bleeding needed further intervention with argon plasma coagulation and/or anti-coagulation drugs in two groups was also compared. RESULTS: Only patients with a definitive cytological or histological diagnosis of lung cancer based on bronchoscopy or other confirmatory techniques were included. Patients were excluded if they had submucosal lesions, extrinsic compressions, pulmonary metastasis of extrapulmonary malignancies or uncommon non-small cell lung carcinoma (NSCLC). A total of 362 patients who met the inclusion criteria were analyzed. Diagnostic yield for pre-biopsy brushing (49.2%, 88/179) was significantly higher than for post-biopsy brushing within the same pre-biopsy brushing group (31.8%, 57/179) (P=0.007) as the intra-group comparison, and significantly higher than for post-biopsy brushing in the post group (30.6%, 56/183) (P<0.001) as the inter-group comparison. No difference in occurrence of treated bleeding for pre- vs. post-biopsy bronchial brushing was found. CONCLUSIONS: Supplementing bronchoscopic forceps biopsy with brushing improves diagnostic yield in lung cancer. In cases of endobronchial exophytic tumors, pre-biopsy brushing appears to be superior to post-biopsy brushing.

Cloning, Expression, Characterization, and Mutagenesis of a Thermostable Exoinulinase From Kluyveromyces cicerisporus.

Inulinase is an enzyme that belongs to glycoside hydrolase family 32. It converts inulin into high-fructose syrups and fructoligosaccharides, both of which are widely used in pharmaceutical and food industries. In this study, the kcINU1 gene (GenBank accession number AF178979) encoding an exoinulinase was cloned from Kluyveromyces cicerisporus CBS4857 and expressed in Pichia pastoris X-33, yielding a maximum of 45.2 ± 0.6 U mL(-1) of inulinase activity of culture supernatant. The expressed inulinase was purified and characterized. The enzyme had an optimum temperature of 55 °C and an optimum pH of 4.5. It had a K m of 0.322 mM and a V max of 4317 µM min(-1) mg(-1) protein when inulin was used as a substrate. It retained nearly 90 % of the maximal activity after pre-incubation at 50 °C for 1 h or at pH ranging from 3.0 to 6.0 at 4 °C for 24 h, demonstrating that KcINU1 was stable at high temperature and low pH. Moreover, we constructed two KcINU1 mutants, Asp30Ala and Glu215Ala, by site-directed mutagenesis and confirmed via zymogram analysis that Asp-30 and Glu-215 of the enzyme were the catalytic active center. The present study has provided important information for understanding the catalytic mechanism of exoinulinase.

Prognostic value of CD133 expression in cancer patients treated with chemoradiotherapy: a meta-analysis.

Many studies evaluated the correlations of CD133 expression with the clinical outcomes in patients treated with chemoradiotherapy (CRT) but yielded controversial results. This meta-analysis was performed to identify the impacts of CD133 expression on the prognosis of cancer patients treated with CRT. Electronic databases updated up to March 2014 were searched to find relevant studies. Relevant literatures without any language restrictions were searched via electronic databases as follows: Web of Science (1945 ~ 2013), the Cochrane Library Database (Issue 12, 2013), PubMed (1966 ~ 2013), EMBASE (1980 ~ 2013), CINAHL (1982 ~ 2013), and the Chinese Biomedical Database (CBM) (1982 ~ 2013). STATA software was used for the current meta-analysis. Hazard ratios (HR) and its corresponding 95% confidence interval (95% CI) were calculated. Six studies were identified with a total of 470 cancer patients treated with CRT. The meta-analysis results showed that CD133-positive patients had poorer overall survival (OS) than that of CD133-negative patients (HR = 2.13, 95% CI = 1.20 ~ 3.07, P < 0.001). Furthermore, CD133-positive patients displayed shorter disease-free survival (DFS) than that of CD133-negative patients (HR = 1.74, 95% CI = 0.08 ~ 3.40, P = 0.039). Ethnicity-stratified analysis indicated that CD133 expression positively correlated with shorter OS among the Japanese, Chinese, and Spanish populations (all P < 0.05). In conclusion, our findings suggest that CD133 expression may be positively correlated with poorer prognosis in cancer patients treated with CRT.

Efficacy of real-time polymerase chain reaction for rapid diagnosis of endobronchial tuberculosis.

OBJECTIVES: The definitive diagnosis of endobronchial tuberculosis (EBTB) is challenging because the disease manifests in various non-specific ways, and acid-fast bacilli (AFB) are often undetectable by sputum smear. The objective of this study was to evaluate the efficacy of real-time PCR of bronchoscopic biopsy specimens for the diagnosis of EBTB. METHODS: Real-time PCR amplification of Mycobacterium tuberculosis DNA in biopsy tissue from EBTB patients was performed prospectively. Diagnostic yields were compared for real-time PCR and for auramine O-stained sputum smears and bronchial brush smears. Whether diagnostic yield depended on bronchoscopic subtype of EBTB was also evaluated. RESULTS: Diagnostic yields were 4.1% (3/74) for sputum smear, 39.2% (29/74) for bronchial brush smear, and 89.2% (66/74) for real-time PCR. Real-time PCR melting curve analysis showed significantly higher yields than did AFB staining of bronchial brush smears for granular and caseating EBTB (p<0.01). CONCLUSIONS: Real-time PCR detection of M. tuberculosis DNA in EBTB biopsy tissue is more sensitive than sputum smear and bronchial brush smear, including at early disease stages. This PCR method may be a useful adjunct to culture- and smear-based techniques to allow more rapid EBTB diagnosis and timelier treatment.

Association of CD14/-260 polymorphism with gastric cancer risk in Highland Tibetans.

AIM: To investigate the relationship between CD14-260 and -651 polymorphisms and the risk of developing gastric cancer. METHODS: DNA was extracted from peripheral blood samples obtained from 225 Tibetans with gastric cancer and 237 healthy Tibetans, and analyzed using the polymerase chain reaction/ligase detection (PCR/LDR) method to determine the genotypes at -260 and -651 loci of the CD14 promoter. The allele frequencies, genotype frequencies, and haplotypes were analyzed for their association with gastric cancer risk using online SHEsis software. The luciferase reporter assay and point mutation analysis were used to construct in vitro plasmids expressing a C/T homozygote at the -260 locus of the CD14 promoter. RESULTS: The frequencies of CC, CT and TT genotypes in the CD14-260 C/T locus in gastric cancer patients were 19.1%, 38.7% and 42.2%, respectively, whereas they were 33.3%, 32.5% and 34.2%, respectively, in healthy control subjects. CT genotype carriers were more frequently found among gastric cancer patients than healthy controls (OR = 2.076; 95%CI: 1.282-3.360). Also, TT genotype carriers were more frequently found among gastric cancer patients (OR = 2.155; 95%CI: 1.340-3.466). Compared to the C allele of CD14/-260, the T allele was associated with an increased risk for gastric cancer (OR = 1.574; 95%CI: 1.121-2.045). Furthermore, the frequencies of CC, CT and TT in the CD14-651 C/T locus in gastric cancer patients were 64.4%, 29.3% and 6.2%, respectively, while they were 56.5%, 35.0% and 8.4%, respectively, in the healthy control subjects (P > 0.05). Data obtained using the luciferase reporter assay showed that the p260T homozygote was associated with greater CD14 promoter activity (P < 0.01). CONCLUSION: CD14/-260 polymorphism is associated with gastric cancer risk in Highland Tibetans and affects CD14 promoter activity, thereby regulating CD14 expression.

Interaction among Caveolin-1 genotypes (rs3807987/rs7804372), H. pylori infection, and risk of gastric cancer in a Chinese population.

Caveolin-1, a candidate tumor suppressor, interacts with a number of transducing molecules and plays a regulatory role in several signaling pathways. Recently, a study revealed that Cav-1 G14713A (rs3807987)/T29107A (rs7804372) polymorphisms might be associated with the susceptibility to certain cancers. In this study, we evaluated the interaction among Caveolin-1 genotypes (rs3807987/rs7804372) and Helicobacter pylori infection and increased risk of gastric cancer among the Chinese population. Blood specimens were collected from 412 gastric cancer cases to 412 noncancer controls between January 2004 and December 2012 in Liaoning Province, China. Caveolin-1 genotypes (rs3807987/rs7804372) were determined by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Enzyme-linked immunosorbent assays were used to measure serum levels of anti-H. pylori IgG. Odds ratio and 95 % confidence interval were calculated using multivariate logistic regression adjusted by sex and age. There were significant differences between gastric cancer and control groups in the distribution of their genotypes and allelic frequencies of the Cav-1 G14713A (rs3807987) and T29107A (rs7804372) polymorphisms, respectively. An elevated risk of gastric cancer was observed in patients with H. pylori infection combined with the Cav-1 G14713A, but not T29107A genotypes. The A allele of G14713A shows an interaction with H. pylori infection that increases the risk of gastric cancer.

[The distribution of Chinese medicine syndrome types in primary liver cancer and their differences of the survival time: a clinical study].

OBJECTIVE: To explore the distribution of Chinese medicine (CM) syndrome types in primary liver cancer (PLC) and their differences of the survival time. METHODS: From May 2007 to March 2009, recruited were 151 PLC inpatients at Department of Tumor, Guangdong Provincial Hospital of Traditional Chinese Medicine. Their survival time were statistically calculated. Patients' average survival time and median survival time were calculated using Kaplan-Meier method. The Log-rank test was used to analyze their differences of survival time among different CM syndrome types. RESULTS: The proportion of CM syndrome types in PLC patients were ranked from high to low as follows: mutual accumulation of dampness and blood stasis syndrome [MADBSS, 43.0% (65/151)], Gan-stagnation Pi-deficiency syndrome [GSPDS, 34.4% (52/151)], qi stagnation blood stasis syndrome [QSBSS, 9.3% (14/151)], retention of damp-heat syndrome [RDHS, 8.6%(13/151)], and Gan-Shen yin deficiency syndrome [GSYDS, 4.6% (7/ 151)]. The median survival time of different CM syndrome types were ranked from longer to shorter as follows: GSPDS (14.77 months), QSBSS (6.13 months), RDHS (5.27 months), MADBSS (4.78 months), and GSYDS (0.80 months). The mean survival times were ranked from longer to shorter as follows: GSPDS (12.40 months), QSBSS (8.84 months), MADBSS (6.99 months), RDHS (7.08 months), and GSYDS (0.72 months). There was statistical difference in the difference of the survival time among different CM syndrome types (P < 0.05). CONCLUSIONS: GSPDS and MADBSS were the most common CM syndrome types in PLC patients. There was difference in the survival time between GSPDS and MADBSS/between RDHS and GSYDS. There was difference in the survival time between MADBSS and GSYDS. Patients of GSPDS might get the best prognosis, while patients of GSYDS might get the poorest prognosis.

The role of E3 ubiquitin ligase Cbl proteins in ß-elemene reversing multi-drug resistance of human gastric adenocarcinoma cells.

Recent studies indicate that ß-elemene, a compound isolated from the Chinese herbal medicine Curcuma wenyujin, is capable of reversing tumor MDR, although the mechanism remains elusive. In this study, ß-Elemene treatment markedly increased the intracellular accumulation of doxorubicin (DOX) and rhodamine 123 in both K562/DNR and SGC7901/ADR cells and significantly inhibited the expression of P-gp. Treatment of SGC7901/ADR cells with ß-elemene led to downregulation of Akt phosphorylation and significant upregulation of the E3 ubiquitin ligases, c-Cbl and Cbl-b. Importantly, ß-elemene significantly enhanced the anti-tumor activity of DOX in nude mice bearing SGC7901/ADR xenografts. Taken together, our results suggest that ß-elemene may target P-gp-overexpressing leukemia and gastric cancer cells to enhance the efficacy of DOX treatment.

Clinical impact of liquid-based cytology test on diagnostic yields from transbronchial needle aspiration.

BACKGROUND AND OBJECTIVE: Transbronchial needle aspiration (TBNA) is a well-established diagnostic method that is underutilized due to the relatively high percentage of non-diagnostic samples and low success rates. This study was designed to evaluate the impact of liquid-based cytology test (LBC) on the diagnostic yield from TBNA. METHODS: Ninety-seven consecutive patients who underwent TBNA due to significant mediastinal adenopathy were enrolled in the study. Each target site was aspirated four times, with the first and third aspirates being prepared for LBC and the second and fourth aspirates being reserved for conventional pick-and-smear (CPS) cytology. RESULTS: Paired aspirates were obtained from 114 target sites, giving a total of 228 test samples from 97 consecutive patients. The overall diagnostic sensitivity of TBNA was 63.6% (56/88). The yields from small cell lung cancers were better than those from non-small cell lung cancers (P < 0.05), and TBNA of subcarinal nodes provided better diagnostic yields (P < 0.05). Nodal diameters > 20 mm on computed tomography were also associated with better yields than nodes with diameters of 10-20 mm (P = 0.001). The diagnostic sensitivity of TBNA was similar for each processing method-59.8% (61/102) for LBC and 64.7% (65/102) for CPS. CONCLUSIONS: LBC was not inferior to CPS with respect to diagnostic yields from TBNA, and enabled efficient pathological evaluation.

ß-Elemene induces apoptosis as well as protective autophagy in human non-small-cell lung cancer A549 cells.

OBJECTIVES: ß-Elemene, a novel traditional Chinese medicine, has been shown to be effective against a wide range of tumours. In this study, the antitumour effect of ß-elemene on human non-small-cell lung cancer (NSCLC) A549 cells and the mechanism involved have been investigated. METHODS: Cell viability and apoptosis were measured by the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively. Protein expression was assayed by Western blotting. Autophagy was evaluated under fluorescence microscopy and transmission electron microscopy. KEY FINDINGS: ß-Elemene inhibited the viability of A549 cells in a dose-dependent manner. This suppression of cell viability was due to the induction of apoptosis. Further study showed that ß-elemene inhibited the activity of the PI3K/Akt/mTOR/p70S6K1 signalling pathway, and at the same time it triggered a robust autophagy. The autophagy was characterized by the accumulation of punctate LC3 dots in the cytoplasm, morphological changes, and the increased levels of LC3-II as well as Atg5-Atg12 conjugated proteins. Inhibition of autophagy with chlorochine significantly enhanced the antitumour effect of ß-elemene. CONCLUSIONS: Our data indicated that ß-elemene inhibited the activity of the PI3K/Akt/mTOR/p70S6K1 signalling pathway in human NSCLC A549 cells, which resulted in apoptosis as well as protective autophagy. A combination of ß-elemene with autophagy inhibitor might be an effective therapeutic option for advanced NSCLC.

[Clinical trial and evaluation on comprehensive treatment on attack in acute stage: report of 522 cases].

OBJECTIVE: To investigate the clinical effect of the comprehensive treatment to acute stage of attack. METHODS: On the basis of the previous observation, the study of the randomization control with general treatment, treatment on acupuncture and western medical treatment were carried out. A comprehensive treatment on overall traditional Chinese medical differentiation according to the superiority of every treatment was assessed and evaluated in 522 patients with attack. RESULTS: The comprehensive treatment of cerebral infarction was superior to the western medicine treatment. General treatment, treatment on traditional Chinese medical differentiation, acupuncture group revealed different improvement on neural function, daily viability, cognitive function in various extent. CONCLUSION: The comprehensive treatment that based on overall traditional Chinese medical differentiation has advantage and characteristic. It has positive combined action to the attack and relevant to clinical setting, easier to popularization and application. Various appraising amount form has different evaluating effects in different stage.

Generation of high affinity human single-chain antibody against PreS1 of hepatitis B virus from immune phage-display antibody library.

BACKGROUND: A single-chain antibody (ScFv) phage display library was created by cloning antigen-binding regions of V(H) (variable domain) and V(L) gene repertoires as fusion proteins with a minor coat protein of filamentous phage, from which high affinity completely humanized ScFv against PreS1 of hepatitis B virus could be screened and characterized. METHODS: A combinatorial library of phage-display human ScFv genes, which were derived from peripheral blood lymphocytes immunized by peptide PreS1 in vitro, was constructed. The library contained 7 x 10(8) clones. RESULTS: After 3 rounds panning, a high affinity (K=10(-7)-10(-8)mol/L) ScFv specific to PreS1 was obtained. Sequence analysis showed that the V(H) belonged to the V(H)4 family and V(lambda) to V(lambda)4. CONCLUSIONS: The described ScFv may provide a more satisfactory therapy. This application further illustrates that the method of in vitro antigen stimulation is expeditious for the source of human immune antibody library.

[Clinical observation on the efficacy enhancing and toxicity attenuating effect of nuzhen yangyin granule to the anti-parkinsonism therapy mainly with Medopa].

OBJECTIVE: To observe and assess the efficacy enhancing and toxicity attenuating effect of Nuzhen Yangyin Granule (NYG) to the anti-parkinsonism (paralysis agitans) therapy with Medopa and Artane. METHODS: Adopting the randomized double-blinded method, the effect of adding NYG to 30 patients with Parkinsonism in the treated group, who already received anti-Parkinsonism treatment but showing decreased response to Medopa and Artane and with obvious adverse reaction, was observed and controlled by 30 patients treated by adding placebo. RESULTS: The total effective rate in the treated group and the control group was 86.7% and 56.7% respectively, the total syndrome improving rate was 90% and 56.7% respectively and the toxicity attenuating rate 90% and 43.3% respectively, comparison between the two groups showed significant difference (P < 0.05 or P < 0.01). NYG also showed markedly effective in reducing the adverse reactions of Medopa and Artane on digestive, neuro-psychiatric and cardiovascular system. CONCLUSION: NYG has obvious efficacy enhancing and toxicity attenuating effects caused by the anti-Parkinsonism treatment with Medopa and Artane.

[Generation of high affinity humanized single-chain antibody against PreS1 of hepatitis B virus from immune phage displayed antibody library].

OBJECTIVE: Screening and characterizing high affinity completely humanized single-chain antibody (ScFv) against PreS1 of hepatitis B virus. METHODS: A combinatorial library of phage-displayed human ScFv, genes of which were derived from peripheral blood lymphocytes immunized by PreS1 of Hepatitis B Virus in vitro, was constructed. The library contained 7 10(8) clones. RESULTS: After 3 rounds panning, a high affinity (K=10(7) to 10(8) mol/L) ScFv specific to PreS1 was obtained. The V(H) belonged to human V(H4) family, and V(1) to V(4) by sequence analysis. CONCLUSION: This study suggests that the method of antigen stimulation in vitro is an expeditious way for the source of human immune antibody. And the ScFv may provide a more satisfactory therapy.

Cloning, expression and purification of gussurobin, a thrombin-like enzyme from the snake venom of Gloydius ussuriensis.

Total RNAs were extracted from the venom gland of the snake Gloydius ussuriensis. The cDNA of gussurobin, a thrombin-like enzyme from Gloydius ussuriensis, was cloned and amplified by RT-PCR. Assay of the nucleotide sequence of the cDNA allowed postulation of the complete amino acid sequence for Gloydius ussuriensis, Chinese Viperdae. Its amino acid sequence exhibits significant homology with that of other snake thrombin-like enzymes. The cDNA of gussurobin was inserted into the vector pPIC 9K and expressed successfully in Pichia pastoris, strain GS115. The recombinant gussurobin was separated and purified from 500 ml culture and showed one band on SDS-PAGE.