Objective: To observe effect of method of regulating Qi to dissipate blood stasis and phlegm on degree of heart failure, ventricular remodeling, disease progression in patients with Qi deficiency and blood stasis type chronic heart failure (CHF).Method: One hundred and thirty-four patients with CHF were randomly divided into control group (62 cases) and observation group (62 cases) by random number table. The patients in control group got spironolactone tablets, 20 mg/time and qd. Benazepril, 20 mg/time and qd. Bisoprolol, 10 mg/time and qd. And digaoxin tablets if necessary. Based on the treatment in control group, patients in observation group additionally received Danshenyin and Xuefu Zhuyutang, 1 dose/day. The treatment course was 3 months in both groups. Before and after treatment, scores of Lee heart failure score were graded, cardiac function classification of the New York Heart Association (NYHA), 6 mins' walking test (6 MWT), scores of Qi deficiency and blood stasis and Minnesota living with heart failure questionnaire (MLHFQ) were evaluated. Echocardiography, left ventricular ejection fraction (LVEF), Left ventricular end-diastolic diameter (LVEDd), left ventricular end-stolic diameter (LVEDs), interventricular septum thickness at end-diastole (IVSd) and left ventricular myocardial mass index (LVMI) were recorded. Levels of matrix metalloprotein-9 (MMP-9), tissue inhibitor of matrix metalloprotease-1 (TIMP-1), transforming growth factor-β₁ (TGF-β₁), N-terminal pro-B-type na-triuretic peptide (NT-proBNP), galectin-3 and copeptin were detected.Result: Ridit analysis showed that after treatment, effect on cardiac function in observation group was better than that in control group (P<0.05). Classification of cardiac function of NYHA was lower than that in control group (P<0.05). The LVEDd, LVEDs and LVMI in observation group were all less than those in control group (P<0.05), while LVEF was higher than that in control group (P<0.05). There was no statistically significant difference in IVSd between two groups. Scores of Lee heart failure score, MLHFQ and Qi deficiency and blood stasis in observation group were lower than those in control group, and distance of 6 mins' walking was larger than that in control group (P<0.01). Levels of serum MMP-9, TGF-β₁, NT-proBNP, galectin-3 and copeptin in observation group were lower than those in control group, while level of TIMP-1 was higher than that in control group (P<0.01).Conclusion: Based on the routine western medicine treatment, additional Danshenyin and Xuefu Zhuyutang can ameliorate symptoms of heart failure, relieve degree of heart failure, improve exercise tolerance and quality of life, inhibit ventricular remodeling, improve cardiac rehabilitation and delay progress of the disease for the CHF patients with Qi deficiency and blood stasis.

OBJECTIVETo map the frequency and types of EGFR gene mutations present in lung cancer tissues. To evaluate the clinical applicability of a novel real-time double-loop probe PCR of which the ADx-EGFR kit is based, and to compare its performance with traditional Sanger DNA sequencing in the detection of somatic mutations of tumor genes.

METHODSA total of 208 formalin-fixed paraffin-embedded (FFPE) tumor samples were tested. Genomic DNA of the tissue samples was extracted and purified, and subjected to both traditional PCR amplification, Sanger sequencing of EGFR gene in exon 18, 19, 20, 21, and ADx's EGFR mutation detection kit. The mutation rates for EGFR gene in exon 18, 19, 20, 21, as well as the frequency of each mutation detected by the two methods, were analyzed.

RESULTSThe traditional Sanger DNA sequencing technique was successfully performed in 196 out of 208 (94.2%) lung cancer samples, and 22 samples (11.2%) showed EGFR gene mutations. ADx-EGFR kit was successfully used in the lung cancers of all of the 208 cases (100.0%), and 40 samples (19.2%) showed mutations. In the lung cancer samples analyzed, mutations were mainly detected in the exon 19 and exon 21 L858R point mutation, i.e. 4.8% (10/208) and 11.6% (23/208) of total mutations, respectively, and the remaining mutations were rare.

CONCLUSIONSThe success rate of ADx-EGFR real-time PCR for formalin-fixed and paraffin-embedded tissues samples is significantly higher than that of Sanger sequencing (P < 0.01). There are significant differences between the two methods. ADx-EGFR real-time PCR shows a much higher successful detection rate and mutation rate of lung cancer tissues compared with that of Sanger sequencing. As a result, the real-time PCR with ADx-EGFR kit is proved to have a good clinical applicability and a strong advantage over the traditional Sanger DNA sequencing. It is an effective and reliable tool for clinical screening of somatic gene mutations in tumors.

DNA Mutational Analysis ; methods ; Exons ; Genes, erbB-1 ; Humans ; Lung Neoplasms ; genetics ; Paraffin Embedding ; Point Mutation ; Real-Time Polymerase Chain Reaction ; methods

OBJECTIVEto map out the frequency and types of K-ras gene mutations present in colorectal and lung cancer patients; to evaluate the clinical applicability of a novel real-time double-loop probe PCR using the ADx-K-ras kit, and to compare its performance with the result by using traditional Sanger DNA sequencing in detection of somatic mutations of the tumor genes.

METHODSa total of 827 formalin-fixed paraffin-embedded (FFPE) blocks including 583 from the colorectal and 244 from the lung cancer patients were assayed. Genomic DNA of the sample tissues was extracted, purified and subjected to PCR amplification of K-ras gene codon 12 and 13 and DNA sequencing was carried on using both the traditional Sanger sequencing method and the ADx's K-ras mutation detection kit, respectively. The mutation rates for K-ras gene at codon 12 and 13, and the mutation frequencies detected by using both methods were analyzed.

RESULTS533 out of 583 (91.4%) colorectal cancer samples and 144 out of 244 lung cancer samples (59.0%) were detected using the traditional Sanger DNA sequencing technique, and 583 out of 583 (100.0%) colorectal plus 244 out of 244(100.0%) lung cancers were detected, respectively by using the ADx-K-ras kit. Of the 583 colorectal cancer samples, 192 (32.9%) showed mutations by using the ADx-K-ras kit in comparing with a result of 160 samples (27.4%) with K-ras gene mutation by using the traditional Sanger DNA sequencing technique. Of the 244 lung cancer samples, 26 (10.7%) showed K-ras gene mutations by using ADx-K-ras kit, while in 144 samples detected by using the traditional Sanger DNA sequencing technique, only 12 samples (8.3%) showed K-ras gene mutations. In colorectal cancer analyzed, GGT→GAT at codon 12 was the most common event with 35.1% (66/188) mutations, followed by GGC→GAC at codon 13 with 26.6% (50/188) and GGT→GTT at codon 12 with 18.6% (35/188), while GGT→GCT at codon12 was the most rare with only 1.6% (3/188) of the total mutation cases. In patients with lung cancer analyzed, GGT→GTT at codon 12 was the most common mutation, accounting for 40.9% (9/22), and GGT→GCT at codon 12 the most rare with only about 4.5% (1/22) of the total mutation cases.

CONCLUSIONSK-ras gene mutations were present in colorectal cases, and significantly more frequent than that in lung cancer. There were significant statistical differences between the two methods. ADx-K-ras real-time PCR showed much higher successful detection rates and mutation ratios compared to Sanger sequencing. As a result, the real-time PCR with ADx-K-ras kit proves to have a good clinical applicability and a strong advantage over the traditional Sanger DNA sequencing. It is a effective and reliable tool for clinical screening of somatic gene mutations in tumors.

Colorectal Neoplasms ; genetics ; Genes, ras ; genetics ; Humans ; Lung Neoplasms ; genetics ; Mutation ; Polymerase Chain Reaction ; methods ; Sequence Analysis, DNA ; methods

Objective To explore the relationship between T lymphocyte subsets and both glioma malignancy and its prognosis, and determine a clinical immunologic index for evaluating preoperative glioma malignancy and its prognosis. Methods The data of 117 inpatients with primary intracranial tumors, including glioma (n=85) and meningioma (n=32), were retrospectively analyzed. Fluorescence-activated cell sorting (FACS) analysis was performed to detect the preoperative contents of T lymphocyte subsets on 32 patients with meningioma and patients with glioma, including 45 high-grade glioma (WHO, grade Ⅲ-Ⅳ) and 40 low-grade glioma (WHO, grade Ⅰ -Ⅱ); and then the differences of their immunologic indexes were analyzed. Based on the detection result of T lymphocyte subsets, patients with glioma were divided into two groups: CD4~+CD8~+<1 and CD4~+CD8~+>1. Follow-up for 3-5 years was performed and the survival difference of these two groups was analyzed. Results Patients with high-grade glioma showed a decreased ratio of CD4~+CD8~+ and an increased value of CD8~+ with significant difference as compared with patients with low-grade glioma (P<0.05); patients with high-grade glioma showed a decreased ratio of CD4~+CD8~+, and an increased value of CD8~+ with statistical significance compared with patients with meningioma (P <0.05); patients with low-grade glioma showed a decreased ratio of CD4~+CD8~+ with statistical significance compared with the patients with meningioma (P<0.05). Patients with glioma showed a decreased ratio of CD4~+CD8~+ and CD4~+, and an increased CD8~+ with statistical significance compared with patients with meningioma (P<0.05). After follow-up for 3-5 years, 48 patients with glioma was found in the CD4~+CD8~+>1 group with 21 death (43.8%) and 31 months as a median survival time; 37 patients with glioma was found in the CD4~+CD8~+<1 group with 23 death (62.2%) and 16 months as a median survival time. The Kaplan-Meier survival curves were analyzed with statistical significance (P<0.05). Conclusion The prognosis is poor in patients with low ratio of CD4~+CD8~+. The preoperative level of T lymphocyte subsets in peripheral blood, correlated to the glioma malignancy, can be considered as an index to evaluate the glioma malignancy and the prognosis in patients with glioma.

OBJECTIVETo evaluate the effectiveness of stroke prevention among high risk population, using Nao'an Capsules.

METHODSParticipants were selected from 696,558 residents in Nanhui, using county of Shanghai city. Individuals aged 35 years old and over with at least one risk factor exposure to stroke, received cerebral vascular hemodynamic examination. 18,271 cases meeting the criteria of cerebral vascular hemodynamic indexes accumulative score below 70 points were defined as individuals with high-risk and targets to receive intervention. According to the willingness of the participants, 10,313 cases received Nao'an Capsules for intensive intervention based on general intervention measures compared to 7958 cases only receiving general intervention. After the implementation of intervention, incidence and mortality rates of stroke and the effectiveness of three-year intervention were studied.

RESULTSIncidence of stroke in Nao'an Capsules group was significantly lower than that of the general intervention (P < 0.01) with 53.8% in males and 58.4% in females. The relative risk (RR) in two gender groups were 0.46 (0.33 - 0.64) and 0.39 (0.30 - 0.50) respectively. Multiple logistic regression analysis indicated that the history of hypertension, accumulative score of cerebral vascular hemodynamic indexes, age, gender and Nao'an Capsules intervention were the variables selected into the equation and significantly related to stroke. Among the variables, Nao'an Capsule was the strongest factor with a RR of 0.41.

CONCLUSIONIncidence of stroke in Nao'an Capsule intervention group was significantly lower than that in the general intervention group after 3 years of intervention, suggesting that Nao'an Capsule intervention was the strongest factor affecting stroke occurrence in individuals at high-risk.

Adult ; Aged ; Capsules ; China ; epidemiology ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Health Education ; Humans ; Incidence ; Logistic Models ; Male ; Middle Aged ; Phytotherapy ; Risk Factors ; Stroke ; epidemiology ; prevention & control

Objective:To compare the effects of different processing techniques on the chemical constituents of Aurantii Fructus for screening the dominant decoction pieces. Method:UPLC-Q/TOF-MS was used to detect the chemical constituents of Aurantii Fructus, chromatography separation was achieved on an ACQUITY UPLC BEH C₁₈ column (2.1 mm×100 mm, 1.7 μm), and gradient elution was performed with 0.1% formic acid aqueous solution (A)-0.1% formic acid acetonitrile solution (B) as mobile phase (0-10 min, 5%-35%B; 10-18 min, 35%-75%B; 18-21 min, 75%-100%B; 21-24 min, 100%B; 24-24.1 min, 100%-5%B; 24.1-28 min, 5%B). Data acquisition was carried out in electrospray ionization (ESI) under the positive ion mode, the scanning range was m/z 50-1 200. The chemical constituents in methanol extract of Aurantii Fructus were identified according to reference substance, relative molecular weight, mass spectrometric cleavage rule and literature information. SIMCA-P 13.0 software was used to establish principal component analysis (PCA) model and partial least squares-discriminant analysis (PLS-DA) model of Aurantii Fructus processed products, PCA score plot, PLS-DA loading plot and variable importance in the protection (VIP) values were obtained to screen the material basis for the main differences before and after processing of Aurantii Fructus. Result:A total of 54 chemical components were identified by UPLC-Q/TOF-MS. PCA indicated that there were significant differences among different groups of Aurantii Fructus processed by different methods. A total of 14 chemical components with VIP value >1 were screened by PLS-DA as the main chemical markers for the differences before and after processing, including hesperidin, poncirin, narirutin, etc. The comprehensive weighted score showed that the content of effective components in Aurantii Fructus processed with honey bran was the highest. Conclusion:The contents of chemical constituents in Aurantii Fructus before and after processing are significantly changed. Flavonoids are the most important compounds to distinguish different processed products of Aurantii Fructus. Aurantii Fructus processed with honey bran is the dominant variety.