The Lived Experience of Frailty in Patients Aged 60 Years and Older with Heart Failure: A Qualitative Study

Purpose@#The prevalence of frailty among patients with heart failure is about 45%. Frailty may result in patients' functional decline, falls, disability, and decreased quality of life. Qualitative studies can explore older patients' perceptions of frailty and help patients cope with it. However, a qualitative approach that explores the experience of frailty in older patients living with heart failure is lacking. This study aimed to explore the lived experience of frailty in older patients with heart failure. @*Methods@#This qualitative study applies Giorgi's phenomenological method. Data were collected from October 2019 to August 2020. Thirteen older patients with heart failure aged at least 60 years were recruited using purposive sampling from a medical center in Taiwan. The participants participated in an in-depth interview using a semistructured interview guide. @*Results@#Seven themes were identified: “being reborn at the end of the road but having difficulty recovering”, “living with a disease with an ineffable feeling”, “feeling like being drained: physical weakness and a dysfunctional body”, “struggling with impaired physical mobility and facing unexpected events”, “suffering from mental exhaustion”, “receiving care from loved ones”, and “turning over a new leaf”. @*Conclusions@#Frailty in older patients with heart failure was obscure and difficult to describe. Frailty could be improved by medical intervention, self-management, and social support but was difficult to reverse. Patients with heart failure should be evaluated for frailty using multidimensional assessment tools at first diagnosis and provided frailty-related information so that patients have proper insight into their disease as early as possible.

Effect of regulating menstruation and promoting pregnancy acupuncture therapy on negative emotion in patients with premature ovarian insufficiency / 中国针灸

OBJECTIVE@#To observe the influence of regulating menstruation and promoting pregnancy acupuncture therapy on negative emotions in patients with premature ovarian insufficiency (POI).@*METHODS@#A total of 60 patients with POI were randomly divided into an acupuncture group and a western medication group, 30 cases in each group. The acupuncture group was treated with regulating menstruation and promoting pregnancy acupuncture therapy at Baihui (GV 20), Shenting (GV 24), Guanyuan (CV 4), Sanyinjiao (SP 6), Shenshu (BL 23), Ciliao (BL 32), etc. once a day, 5 times a week for 3 months. The western medication group was treated by oral administration of climen. The drug was given 1 tablet a day for 21 days and was stopped for 1 week as a course. The treatment was required 3 consecutive courses. The self-rating anxiety scale (SAS) score, modified Kupperman index (KI) score, agitated and depressive symptom scores in KI and serum level of follicle stimulating hormone (FSH) before and after treatment were compared between the two groups.@*RESULTS@#After treatment, the SAS scores, KI scores and serum levels of FSH in the two groups and the scores of agitated and depressive symptom in the acupuncture group were lower than those before treatment (@*CONCLUSION@#Regulating menstruation and promoting pregnancy acupuncture therapy can effectively improve the negative emotions of patients with POI and reduce serum level of FSH .

A new research paradigm in modernization of traditional Chinese medicine: single cell pharmacology / 药学学报

Single cell "omics" technology enables the capture of genome, transcriptome, proteome and other omics information in a high-throughput and unbiased manner at single-cell resolution, allowing the characterization of the functional state of individual cells to reveal their heterogeneity and differential responses to drug treatment. This technology has wide application in pharmacological research, facilitating drug screening, efficacy evaluation, and mechanistic studies. We envision that, in the field of traditional Chinese medicine (TCM), single cell omics technology can be applied in the identification of active ingredients and drug targets, and elucidation of drug mechanism of action. In this article, we briefly introduce the single cell omics technology - particularly single cell transcriptome sequencing, and review its application in the field of modern drug research. Based on that, we propose the concept of "single cell pharmacology" and articulate how it can be applied to transform the pharmacological research of TCM and promote TCM modernization.

Challenges and countermeasures of thoracic surgery in the epidemic of novel coronavirus pneumonia / 中国胸心血管外科临床杂志

@#Since December 2019, a novel coronavirus (2019-nCoV, SARS-CoV-2) pneumonia (COVID-19) outbreak has occurred in Wuhan, Hubei Province, and the epidemic situation has continued to spread. Such cases have also been found in other parts of the country. The spread of the novel coronavirus pneumonia epidemic has brought great challenges to the clinical practice of thoracic surgery. Outpatient clinics need to strengthen the differential diagnosis of ground glass opacity and pulmonary plaque shadows. During the epidemic, surgical indications are strictly controlled, and selective surgery is postponed. Patients planning to undergo a limited period of surgery should be quarantined for 2 weeks and have a nucleic acid test when necessary before surgery. For patients who are planning to undergo emergency surgery, nucleic acid testing should be carried out before surgery, and three-level protection should be performed during surgery. Patients who are planning to undergo emergency surgery in the epidemic area should be confirmed with or without novel coronavirus pneumonia before operation, and perform nucleic acid test if necessary. Surgical disinfection and isolation measures should be strictly carried out. Among postoperative patients, cases with new coronavirus infection were actively investigated. For the rescue of patients with novel coronavirus infection, attention needs to be paid to prevention and treatment and related complications, including mechanical ventilation-related pneumothorax or mediastinal emphysema, and injury after tracheal intubation.

Effect of vinegar processing on esculentosides in n-BuOH fraction and main toxic components in Phytolaccae Radix / 中国中药杂志

This study was to investigate the effect of vinegar processing on esculentosides in n-BuOH fraction and the contents of the main toxic components esculentoside B (EsB) and esculentoside C (EsC) in Phytolaccae Radix pieces. n-BuOH fraction of Phytolaccae Radix pieces was processed with vinegar according to the processing method in Chinese Pharmacopoeia. HPLC-MS-MS was adopted to analyze the esculentosides composition changes in n-BuOH fraction before and after vinegar processing. HPLC-ELSD was used to detect EsC and EsB contents in raw and vinegar processed Phytolaccae Radix pieces, and investigate the content changes before and after vinegar processing. Results showed that the esculentosides contents in n-BuOH fraction were significantly decreased except esculentoside A (EsA); there were significant changes in saponins compositions, but no new compounds were generated in n-BuOH fraction after vinegar processing. The contents of EsC and EsB were 0.12% and 0.20% respectively in raw Phytolaccae Radix, and decreased to 0.048% and 0.094% accordingly after vinegar processing. It showed that vinegar processing could significantly change the composition of esculentosides in n-BuOH fraction from Phytolaccae Radix and reduce the contents of toxic components EsC and EsB, indicating the scientificity of vinegar processing for Phytolaccae Radix.

Excimer laser atherectomy combined with drug-eluting balloon angioplasty for the treatment of chronic ischemia of lower limbs: preliminary results in three patients / 介入放射学杂志

Objective To evaluated the safety and feasibility of excimer laser atherectomy (ELA) combined with drug-eluting balloon angioplasty in treating chronic ischemia of lower limbs.Methods ELA combined with paclitaxel-eluting balloon angioplasty was adopted to treat chronic ischemia of lower limbs caused by arteriosclerosis occlusive disease of lower extremity in three patients.All three patients had arteriosclerosis occlusive disease of superficial femoral artery;in two of them the disease was primary occlusive lesion and in another patient the disease was in-stent re-occlusion lesion after sten implantation.Results After the treatment,the blood flow in the diseased arteries was unobstructed,the blood supply of the lower limbs was obviously improved.No procedure-related complications occurred.Two weeks after the treatment,no recurrence of ischemic symptoms was observed,the blood flow in superficial femoral artery kept unobstructed.The patients recovered smoothly.Conclusion For the treatment of chronic ischemia of lower limbs,which are caused by the primary arteriosclerosis occlusive disease of lower extremity or by the in-stent re-occlusion lesion after sten implantation,ELA combined with paclitaxel-eluting balloon angioplasty is clinically safe and feasible,although its long-term effect needs to be clarified with more studies.

Evaluation of the learning curve of laparoscopic choledochal cyst excision and Roux-en-Y hepaticojejunostomy / 中国内镜杂志

Objective To evaluate the outcome of the laparoscopic choledochal cyst excision and Roux-en-Y hepaticojejunostomy. Then define the characteristics of the learning curve of this procedure. Methods A prospectively collected database comprising all medical records of the first 79 consecutive patients underwent laparoscopic choledochal cyst excision and Roux-en-Y hepaticojejunostomy performed by one surgeon was studied. The patients were divided into seven groups (13 cases in last group) by operative sequence. Data on patients' demographics, clinical and outcome variables including operative duration, conversion to open surgery, complications, and length of hospital stay were analyzed. The learning curve for the laparoscopic choledochal cyst excision and Roux-en-Y hepaticojejunostomy was established through the moving average and ANOVA methods. Results Comparing the early with the late experiences (33 v. 46 cases), the surgeon-specific outcomes significantly improved in terms of operating times [(292.7 ± 29.8) vs (215.3 ± 10.2) min, P < 0.05], There was four patients converted to open surgery in the early of experiences.the late experiences was five case, it was no statistical significance. significant differences were not shown in the hospitalization period and infectivity complications. Conclusion Operative duration can be reduced with increasing experience of laparoscopic choledochal cyst excision and Roux-en-Y hepaticojejunostomy, In this study, the learning curve for a laparoscopic choledochal cyst excision and Roux-en-Y hepaticojejunostomy was about thirty-three cases.

Intestinal toxicity of n-BuOH fraction from Phytolacca Radix before and after being processed with vinegar / 中国中药杂志

To research the intestinal toxicity of n-BuOH fraction in Phytolacca Radix before and after being processed with vinegar. Toxic n-BuOH fractions were separated from Phytolacca Radix. In the animal model, the level of intestinal edema, water content of intestine and stool, IC₅₀ values of HT-29 and IEC-6 were detected with MTT method to compare the changes in toxicity of n-BuOH fractions from Phytolacca Radix before and after being processed with vinegar. n-BuOH fractions of Phytolacca Radix could cause intestinal edema in mice, increase the edema of duodenum, jejunum and the water content in stool, inhibit the proliferation of HT-29 cells and IEC-6 cells, indicating its intestinal toxicity, with HT-29 IC₅₀ at 14.59 mg•L⁻¹ and IEC-6 IC₅₀ at 43.77 mg•L⁻¹. After being processed with vinegar, the level of intestinal edema, edema of duodenum and jejunum and the water content in stool and inhibition ratio of cells line were reduced, with HT-29 IC₅₀ at 58.51 mg•L⁻¹ and IEC-6 IC₅₀ at 84.37 mg•L⁻¹. After being processed with vinegar, the toxicity of n-BuOH fractions from Phytolacca Radix decreased obviously.

Antagonism mechanism of gingerols against inflammatory effect of toxic raphides from Pinella pedatisecta / 中国中药杂志

This study was to investigate the mechanism of gingerols antagonizing the inflammatory effect of toxic raphides from Pinella pedatisecta. Mice peritonitis models induced by toxic raphides from P. pedatisecta were applied to observe the effect of gingerols on inflammatory mediators PGE2 in the exudates of abdominal inflammation in mice; rats peritoneal macrophage in vitro culture models were adopted to study the anti-inflammatory effects of gingerol against toxic raphides, with TNF-α and IL-1β in supernatant as indexes. Scanning electron microscopy was used to observe the changes in surface morphology of macrophages treated by raphides and gingerols. Macrophages-neutrophils co-cultured models were used to study the antagonism of gingerols against the effect of toxic raphides' stimulation on neutrophils migration. Results showed that gingerols could significantly inhibit the production of PGE2 in the exudates of abdominal inflammation induced by toxic raphides from P. pedatisecta in mice. Gingerols could significantly inhibit the toxic raphides from P. pedatisecta to induce the release of inflammatory factors, with certain dose dependence. Scanning electron microscopy showed that gingerols could significantly inhibit phagocytosis of macrophages, cytomembrane injury, and neutrophils migration induced by toxic raphides from P. pedatisecta. The results showed that the antagonism mechanism of gingerols against the toxic raphides from P. pedatisecta may be associated with inhibiting the pro-inflammatory toxicity including macrophage activation, inflammatory factors release, and neutrophils migration.

Totoxicity fraction from Euphorbia pekinensis and composition change after vinegar processing / 中国中药杂志

To look for the toxicity fraction of Euphorbia pekinensis and discuss the vinegar processing mechanism. The level of intestinal edema, water content of intestine and stool, IC50 values of IEC-6 were applied to evaluate the toxicity of different fractions. RT-PCR was employed for detecting AQP1, AQP3 mRNA expression. The petroleum ether (PE) fraction and ethyl acetate (EtOAc) fraction could significant cause intestinal edema in mice, increase the water content of duodenum, colon and stool, inhibited the mRNA expression of AQP1 and increased the mRNA level of AQP3 in colon, and the petroleum ether (PE) fraction was more poisonous. After the petroleum ether (PE) fraction was processed with vinegar, the level of intestinal edema, water content of duodenum, colon, stool and inhibition ratio of cells line were reduced. And we compared the composition change after vinegar processing, finding that the conpekinensis.

Advance in oral vaccine / 中国药学杂志

Oral vaccine can stimulate the immunity of intestinal mucosa effectively after entering the gastrointestinal orally. It is a more ideal way of immunity compared with the traditional injection vaccine, since oral vaccine delivery mode is simple and quick while the results of immunity is stable and durable. There are many types of oral vaccines, including live attenuated vaccine, DNA vaccine and transgenic plant vaccine. Oral vaccine vehicle has more types, which not only can enhance the body to absorb the antigen but also can enhance the immune effect of the protective antigen effectively. In this paper, the mechanism about immunologic effect and recent advance on the research progression of oral vaccine are reviewed, what would provide ideas for related basic research.

Expression and clinical significance of circadian gene Per2 in non-small cell lung cancer / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To detect the expression of Per2 in non-small cell lung cancer (NSCLC), and analyze its clinical significance.</p><p><b>METHODS</b>The expression of Per2 was determined in 60 NSCLC and 20 normal lung tissues by immunohistochemical assay, and the relationship between Per2 expression and clinicopathological features was analyzed.</p><p><b>RESULTS</b>The positive expression rates of Per2 in NSCLC and normal lung tissues were 71.7% and 95.0%, respectively (P < 0.05). The expression of Per2 in NSCLC was correlated with pathological differentiation and TNM stage (P < 0.05).</p><p><b>CONCLUSION</b>The expression of Per2 in NSCLC is decreased. The negative expression of Per2 may contribute to the development and invasion in NSCLC.</p>

Value of serum P53 antibody as a tumor marker surveillance of colorectal cancer within follow-up after curative resection / 中华胃肠外科杂志

P53 gene mutations and the abnormal P53 protein can introduce the production to P53 antibody. A large number of studies showed that serum levels of P53 antibody had the correlation with the prognosis of patients with different cancers, the lymph node invasion and metastasis of colorectal cancer, and its recurrence after the curative resection. And it is possible for its application in predicting the early recurrence and metastasis in colorectal cancer after the curative resection.However, there are still a lot of work needed to be done before its use in the clinical settings.

HIWI expression profile in cancer cells and its prognostic value for patients with colorectal cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>HIWI is a member of PIWI gene family and its expression is found in various tumors, indicating that it may play a pivotal role in tumor development. This study was designated to examine HIWI protein expression profile in several cancer cell lines and its prognostic value for patients with colorectal cancer.</p><p><b>METHODS</b>Totally 270 patients who underwent surgical resection of primary colorectal cancer between January 1999 and December 2002 with a median follow-up time of 33 months were registered in the study. Formalin-fixed and paraffin-embedded specimens from these patients and 236 matched adjacent non-cancerous normal colorectal tissues were collected. Anti-HIWI monoclonal antibodies were generated and used for evaluating HIWI protein expression. χ(2) tests were conducted to determine the association between HIWI expression and the other variables. Survival curves were estimated using the Kaplan-Meier method and compared by the log-rank test. Multivariate analysis was performed by using the Cox regression model.</p><p><b>RESULTS</b>By generating antibodies specific for HIWI, we examined HIWI protein expression in several cancer cell lines and demonstrated positive expression of HIWI in 69 out of 270 (25.6%) colorectal cancer tissues; 15 of 236 (6.4%) matched adjacent non-cancerous tissues were also positive for HIWI. Patients with positive HIWI expression in adjacent non-cancerous tissue had statistically lower overall survival (OS) and disease free survival (DFS) compared with negative patients (OS: 10.4% vs. 55.5%, P = 0.009; DFS: 10.4% vs. 55.1%, P = 0.015). For early stage group (stages I and II), patients with positive HIWI expression had significantly lower OS and DFS (OS: 57.4% vs. 79.5%, P = 0.014; DFS: 56.7% vs. 80.5%, P = 0.010). In lymph node negative group, patients with positive HIWI expression had statistically lower OS and DFS (OS: 53.0% vs. 73.5%, P = 0.037; DFS: 52.2% vs. 74.6%, P = 0.025). Multivariate analysis revealed that HIWI over-expression was a significant prognostic factor for OS (95%CI: 1.132 - 2.479, P = 0.010).</p><p><b>CONCLUSION</b>HIWI could be a potential prognostic biomarker for the patients with colorectal cancer, especially for those at early stages or without lymph node metastasis.</p>

Application of biotherapy in recurrent or metastatic gastric cancer / 中华胃肠外科杂志

Post-operative recurrence and metastasis remain the leading causes of death for patients with gastric cancer. The major determinants of recurrence and metastasis are the biological characteristics of cancer cells and the immunological status of the patients. In recent years, due to the target-specificity, biotherapy has yielded efficacious responses in diverse clinical applications for cancer treatment, partially for the treatment of recurrence and metastasis of gastric cancer. However, because of the high diversities in clinical manifestations, patients' condition, and tumor's characteristics, there is no ideal strategy of biotherapy established for the prevention and treatment of recurrence and metastasis in gastric cancer. Therefore, a lot of work need to be done in basic research and clinical trial to make the biotherapy effective in treatment of gastric cancer recurrence.

Radiofrequency treatment on respiratory symptoms due to gastroesophageal reflux disease / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Diagnosis and treatment for respiratory symptoms (RSs) of gastroesophageal reflux disease (GERD) is more difficult than that for common esophageal symptoms. The goal of this study was to evaluate the efficacy and safety of radiofrequency (RF) treatment on RSs of GERD in a preliminary 12-month follow-up observation.</p><p><b>METHODS</b>From April 2006 to October 2008, 505 GERD patients with mainly respiratory presentations such as wheezing, chronic cough or hoarseness, were treated by endoscopic RF. A questionnaire was completed before and after treatment, using a six-point scale ranging from 0 to 5 to assess symptom severity and frequency. The symptom score was the sum of frequency and severity.</p><p><b>RESULTS</b>Symptom scores were significantly improved at the end of the follow-up period. The mean heartburn score decreased from 5.31 to 1.79. The mean regurgitation score decreased from 5.02 to 1.64; mean cough score decreased from 6.77 to 2.85; mean wheezing score decreased from 7.83 to 3.07; and mean hoarseness score decreased from 5.13 to 1.81 (P < 0.01). No major complications or deaths occurred. Minor complications included temporary post-procedural retrosternal unease or pain (n = 106; 21.0%), mild fever (n = 86; 17.0%), transient nausea/vomiting (n = 97; 19.2%), and transient dysphagia (n = 42; 9.3%). Thirty-five (6.9%) patients had recurrence of symptoms. Endoscopic RF treatment was repeated in six patients, and laparoscopic fundoplication was performed in seven.</p><p><b>CONCLUSION</b>Endoscopic RF is an effective and safe means to treat RSs in patients with GERD.</p>

miRNA changes in the reverse remodeling heart of rats / 中华心血管病杂志

<p><b>OBJECTIVE</b>To establish a reverse remodeling heart model in rats and observe collagen and TGF-β expression and relevant microRNAs changes during reverse remodeling.</p><p><b>METHODS</b>Lewis rats were divided into four groups including sham (NL, n = 10), abdominal aortic constriction (AAC, n = 10), heterotopic transplantation of abdominal aortic constriction (AAC-HT, n = 9) and heterotopic transplantation of normal heart (HT, n = 8). Left ventricular wall thickness and LV cavity were measured by echocardiography. The cardiomyocyte cross-sectional area (CSA) was determined on HE stained sections. Immunohistochemical and qRT-PCR were used to detect collagen and TGF-β expressions. miRNAs were detected by MicroRNA microarray.</p><p><b>RESULTS</b>Heart weight, left ventricular wall thickness and CSA were significantly increased in AAC hearts compared to those in the NL and AAC-HT hearts. The collagen and TGF-β were increased in AAC hearts and further increased in AAC-HT hearts. miRNA microarray evidenced more than two folds changes on 82 miRNAs compared to NL (10 in AAC, 32 in AAC-HT and 40 in HT).</p><p><b>CONCLUSION</b>Rat abdominal aortic constriction and heterotopic transplantation could be used as a reverse remodeling heart model and significant collagen and TGF-β as well microRNA expression changes were evidenced in this model.</p>

Genetic variants in three genes and smoking show strong associations with susceptibility to exudative age-related macular degeneration in a Chinese population / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The present study was undertaken to replicate the associations of representative polymorphisms in three genes (complement factor H (CFH), complement factor B (BF) and HtrA serine peptidase 1 (HTRA1)) with exudative age-related macular degeneration (AMD) in a Han Chinese population, and to test if the modifiable environmental factors affect AMD susceptibility associated with different type of genotype in these genes.</p><p><b>METHODS</b>An age, gender and ethnicity matched case-control study was conducted to genotype the representative single neucleotide polymorphisms (SNPs) loci including rs1061170 and rs1410996 in CFH, rs641153 and rs4151667 in BF and rs11200638 in HTRA1 gene in 144 exudative AMD patients and 126 normal controls using PCR-RFLP and direct resequencing. The demographic characteristics and behavioral risk factors were also recorded. Allelic and genotypic associations for individual SNP and joint associations with two loci were performed. The gene-gene and gene-environment interactions were analyzed using multivariate non-conditional Logistic regression analysis.</p><p><b>RESULTS</b>The C risk allele frequencies for CFH Y402H (rs1061170) in cases and controls were 12.5% and 5.4% respectively, which were much lower than those in Caucasians (P < 0.001). Compared with TT homozygous genotype, the CT heterozygous genotype was positively associated with AMD with odds ratio (OR) of 3.23 (1.36 - 5.07). However, the population attributable risk (PAR) of C allele was only 3.3% (1.4% - 4.3%). rs1410996 was also associated with AMD independent of Y402H. The ORs of exudative AMD for individuals carrying one copy risk allele and two copy risk alleles were 2.57 (1.21 - 5.45) and 4.76 (2.15 - 10.55) respectively, with correspondent PARs of 28.3% (2.0% - 40.5%) and 38.2% (21.8% - 45.4%). rs11200638 in HTRA1 was another susceptible locus for AMD and the risk homozygotes were significantly susceptible for exudutive AMD (OR = 3.98, 1.88 - 8.43) with PAR of 38.9% (24.3% - 45.8%). Education status and cigarette smoking were also related to exudative AMD. After controlling for environmental risk factors, CFH and HTRA1 SNPs were independently associated with exudative AMD, with OR of 3.50 (1.45 - 8.45) for CT genotype in Y402H, 3.34 (1.33 - 8.36) for GG genotype in rs1410996 and 3.85 (1.58 - 9.42) for AA genotype in rs11200638 respectively. The interaction analysis between gene and environmental factors showed that smoking synergistically increased susceptibility of AMD for heterozygotes of rs1410996, with OR(interaction) of 7.33 (P(interaction) = 0.029).</p><p><b>CONCLUSIONS</b>In a Han Chinese population, CFH and HTRA1 polymorphisms appear to be independently and possibly additively hereditary contributors to exudative AMD. Y402H polymorphism conferred a significant but relatively lower contribution in Chinese than in Caucasians with a low frequency of risk allele. The gene-environment interaction may be a best way to encourage those with a high genetic risk to prevent AMD by avoiding modifiable factors until there is effective treatment for AMD.</p>

A20 inhibits human salivary adenoid cystic carcinoma cells invasion via blocking nuclear factor-kappaB activation / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>A20, also known as tumor necrosis factor alpha induced protein 3 (TNFaip3), is a cytoplasmic zinc finger protein that inhibits nuclear factor kappa-B (NF-kappaB) activity and prevents tumor necrosis factor (TNF)-mediated programmed cell death. NF-kappaB is a transcription factor that regulates expression of genes involved in cell proliferation, cell survival and anti-apoptosis. Several studies have implicated that the NF-kappaB signal pathway is associated with angiogenesis and clinico-pathological process of adenoid cystic carcinoma (ACC) of the salivary glands.</p><p><b>METHODS</b>The ability of overexpression of A20 to influence the biological behavior and invasion of ACC cells was examined. The cells were stably transfected with full-length A20 cDNA. Stable gene transfer was verified by realtime-polymerase chain reaction (PCR) and Western blot analysis. The change of cell biological behavior was examined by methyl thiazolyl tetrazolium (MTT) and NF-kappaB luciferase reporter assay and the invasion of the cells was examined by a Matrigel invasion chamber.</p><p><b>RESULTS</b>pEGPFN3-A20 gene was stably transferred into ACC-2 cells and overexpressed. When cells were treated with TNFalpha, the NF-kappaB activity of ACC-2-A20 cells could be down-regulated about 46.32% in contrast to ACC-2-GFP cells (P < 0.05). A20 potently inhibited growth of A20 transfectant ACC-2-A20 compared with control vector transfected groups and the ACC-2 empty control group (P < 0.05). The ACC-2-A20 cells showed significantly reduced ability to invade through Matrigei-coated filters compared to ACC-2-GFP and ACC-2 cells. The inhibition rate was up to 71.05% (P < 0.05).</p><p><b>CONCLUSIONS</b>A20 gene transfer is associated with decreased tumor invasion, in part via the down-regulation of NF-kappaB expression, providing evidence for a potential application of A20 in designing a treatment modality for salivary gland cancers such as ACC.</p>

Effect of trastuzumab on tumor cell lines shedding high or low level of HER-2 ECD / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To examine the effect of trastuzumab on cell proliferation, colony formation and changes of HER-2 proteins in human breast cancer cell line SKBR3 and human ovarian cancer cell line SKOV3 cells which overexpress p185 HER-2 but shed high or low HER-2 extracellular domain (ECD) levels.</p><p><b>METHODS</b>SKBR3 cells and SKOV3 cells were treated with or without trastuzumab. Cell number and the rate of colony formation were calculated. Western blot analysis was used to detect p185 HER-2, HER-2 ECD and phospho-HER-2. Two-site ELISA assay was used for the detection of HER-2 ECD.</p><p><b>RESULTS</b>Trastuzumab inhibited cell proliferation, colony formation, and decreased or eliminated the levels of two uncharacterized phospho-proteins (molar weight about 90 000 and 40 000) in SKBR3 cells shedding high level of HER-2 ECD expression. These responses were not observed in SKOV3 cells shedding low level of HER-2 ECD expression. But total p185, phospho-p185 and phospho-p95 proteins did not appear to change in SKBR3 and SKOV3 cells after treatment with trastuzumab. Trastuzumab reacts not only with proteolytic cleavage HER-2 ECD containing HER-2 ECD I , II , III and IV subdomains of p185 HER-2 extracellular domain, but also with the secreted autoinhibitor p68/ECD III a specifying 340 residues, identical to subdomains I and II from the extracellular domain of p185 HER-2, followed by a unique C-terminal sequence of 79 aa encoded by intron 8, which suggested that there may be a trastuzumab binding site on p68/ECD III a protein. Comparing with HER-2 ECD levels of the same number of SKBR3 cells, there was no significant decrease of HER-2 ECD shedding level after treatment with or without trastuzumab for 4 days in serum-free medium.</p><p><b>CONCLUSION</b>Antitumor effects of trastuzumab may be related to the two uncharacterized phospho-p90 and/or phospho-p40 proteins. There is probably a trastuzumab epitope on p68/ECD III a. The decrease of HER-2 ECD levels may be positively correlated with the number of SKBR3 cells.</p>