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Puerarin (Pue), known as a phytoestrogen, has salient bioactivities and is promising against cardiovascular diseases. This article summarizes the underlying molecular mechanisms of Pue in treating cardiovascular diseases, especially regulating the intracellular signal transduction, influencing ion channels, modulating the expression of microRNA, and impacting on the autophagy, which are mainly involved in the inflammatory signaling pathways, fatty acid/lipid metabolism, oxidative stress, apoptosis, and the like. The protective effect of Pue against cardiovascular diseases mainly involves attenuating the myocardial injury and decreasing the myocardial fibrosis, improving the myocardial ischemia/reperfusion injury, as well as inhibiting the myocardial hypertrophy and atherosclerosis. The molecular mechanisms of Pue's cardiovascular protective effects for the first time and comment on the state-of-the-art research methods and principles of Pue's regulation of small molecules were reviewed, so as to provide the rationale for its basic research and clinical applications.
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Objective:To investigate the relationship between BRAF V600E gene mutation and ultrasonography manifestations as well as the lesion invasiveness in thyroid cancer.Methods:A total of 153 patients pathologically diagnosed as thyroid cancer after surgery who underwent thyroidectomy in Shanxi Provincial Cancer Hospital from January 2018 to October 2019 were selected, including 146 cases of papillary thyroid carcinoma. Ultrasonography was performed before operation. Paraffin embedded tissue after operation was used to detect BRAF V600E gene mutation. According to the results of BRAF V600E gene detection, patients were divided into mutation group and non-mutation group. The ultrasonic characteristics of the two groups were compared. The relationship of BRAF V600E gene mutation with ultrasonic characteristics, clinicopathological characteristics as well as cervical lymph node metastasis was analyzed by using logistic regression.Results:There were 130 cases (85.0%) of BRAF V600E gene mutation and 23 cases of BRAF V600E gene non-mutation in 153 patients with thyroid cancer. Among 146 cases with papillary thyroid carcinoma, there were 128 cases (87.7%) of BRAF V600E gene mutation. The percentage of patients with the unclear boundary between thyroid lesions and capsule in BRAF V600E gene mutation group was higher than that of patients in non-mutation group, and the difference was statistically significant [46.9% (60/128) vs. 11.1% (2/18), χ 2 = 8.261, P = 0.004]. There were no significant differences in age, gender, nodule long diameter, aspect ratio, nodal location, internal calcification, internal echo, echo uniformity, cystic solid, nodal shape, boundary clarity, blood flow signal, the number of tumor site, lymph node metastasis and nodular goiter between BRAF gene mutation group and non-mutation group (all P > 0.05). The results of logistic regression analysis showed that only the boundary clarity between thyroid lesions and capsule was an independent influencing factor of BRAF V600E gene mutation ( OR = 14.400, 95% CI 1.847-112.246, P = 0.011), tumor lesion size was an independent influencing factor of cervical lymph node metastasis in papillary thyroid carcinoma ( OR = 2.714, 95% CI 1.335-5.517, P = 0.006). Conclusions:In papillary thyroid carcinoma, BRAF V600E gene mutation is related with lesion and the unclear boundary between the tumor and capsule, but not related with lymph node metastasis. The size of the tumor lesion is associated with lymph node metastasis.
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Objective To detect C-met protein expression and gene amplification in lung adenocarcinoma, and to analyze their relationship with epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) resistance and prognosis. Methods A total of 120 cases of lung adenocarcinoma diagnosed in Shanxi Provincial Cancer Hospital from January 2011 to May 2013 were selected. The expressions of C-met protein and C-met gene amplification were conducted by immunohistochemistry (IHC) method and fluorescence in situ hybridization (FISH), and all patients were followed up. The relationship between the expression of C-met protein and gene amplification with clinicopathological features and EGFR-TKI resistance and prognosis were analyzed. Results The high expression of C-met protein and gene amplification in 120 tissues were 17.5 % (21/120), 10.83 % (13/120). Of the 80 patients treated with EGFR-TKI, the incidence of C-met protein high expression was 30.43 % (14/46) in patients with drug resistance, which was significantly higher than that in patients without drug resistance (11.76 %, 4/34), the difference was statistically significant (χ2= 3.908, P= 0.048). The rate of C-met gene amplification was 19.57 % (9/46) in patients with drug resistance,which was significantly higher than that in patients without drug resistance (2.94 %, 1/34) the difference was statistically significant (P= 0.038). The expression of C-met protein in 46 patients with drug resistance was positively correlated with gene amplification (r= 0.388, P= 0.008), but in 40 patients without TKI, the expression of C-met protein was not correlated with gene amplification (r=0.279, P=0.081). The high expression of C-met protein was correlated with age, pathological grade and clinical stage (all P<0.05), while C-met gene amplification was related to clinical stage (P=0.036). Cox regression analysis suggested that C-met gene amplification was an independent prognostic factor (P= 0.034). Conclusions C-met protein expression and gene amplification are risk factors for EGFR-TKI resistance. C-met gene amplification suggests poor prognosis, and can be used as an independent factor for prognostic evaluation.
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Objective To explore the correlation between anaplastic lymphoma kinase (ALK) fusion gene detected by immunohistochemistry (IHC) and reverse transcription-polymerase chain reaction (RT-PCR) in non-small cell lung cancer. Methods The ALK fusion protein/gene in 71 patients of NSCLC which was detected both by IHC (1A4/1H7) and RT-PCR were retrospective studies, and the 2 methods were compared. Results Among the 71 NSCLC patients, the ALK fusion protein positive was in 21 cases and negative was in 50 cases by IHC detected, while the ALK fusion gene positive was in 12 cases and negative was in 59 cases by RT-PCR. The ALK fusion genes detected by RT-PCR were all negative when IHC negative and IHC 1+. All patients with IHC 2+ and IHC 3+ were confirmed ALK fusion genes positive with RT-PCR. The positive rate of ALK fusion protein detected by IHC in large surgical specimens was 28.95%(11/38), and the positive rate of ALK fusion protein detected by IHC in small biopsy specimen was 30.30%(10/33). The positive rate of ALK fusion gene detected by RT-PCR in large surgical specimens was 18.42%(7/38), and the positive rate of ALK fusion gene detected by RT-PCR in small biopsy specimen was 15.15% (5/33). Conclusions Although the ALK fusion protein detected by IHC may have certain false positive, IHC is highly consistent with RT-PCR in IHC 2+and IHC 3+ cases. The combination of IHC and RT-PCR can be used to ALK fusion gene positive NSCLC screening and diagnosis. The small biopsy specimen is also good material for ALK detection, when the surgical specimen can not be got from patients.
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Objective:To establish a fibrin zymography method for identifying the active proteins in lumbrokinase,and investigate the production difference and batch consistency of 5 different manufacturers. Methods:Fibrin zymography was used with the final con-centration of fibrinogen of 5 × 10 - 4 g·ml - 1 ,renature time of 30 min in 2. 5% Triton-x-100,incubation time of 30 min in PBS buffer (pH = 7. 4)at 37℃ and the protein concentration in the sample of 5. 0-37. 5 μg. Results:The sensitivity of the method was high,and the molecular weight distribution of active protein bands for the samples from five manufacturers was between 15KD and 40KD with 6 common active protein bands. The zymography of the samples from the five manufacturers had slight difference,while various batches of the samples from the same manufacturer showed no difference. Conclusion:The method is special. It can reflect molecular weight distribution and species of active protein,batch consistency and production process stability. It is easy to be standardized and suitable for the identification of lumbrokinase,which can lay foundation for the quality consistency evaluation of marketed products.
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Objective:To establish a Crownpak CR( +)-HPLC method for the determination of the content and related substance of valacyclovir hydrochloride dispersible tablets. Methods: A Crownpak CR( +) [4. 0 mm × 150 mm,5 μm,DAICEL CROWNPAK CR( +)] column was used,and the mobile phase was 0. 1% perchloric acid in water. The flow rate was 0. 75 ml·min-1 and the de-tection wavelength was 255 nm. Results: A good linear range of valacyclovir hydrochloride was 11. 25-180. 00 μg · ml-1 ( r =1. 000 0), and the average recovery was 99. 0%(RSD=0. 8%, n=9). A good linear range of alacyclovir was 0. 2-50μg·ml-1(r=1. 000 0), and the average recovery was 99. 3%(RSD=0. 6%, n=9). The content of the tablets from two pharmaceutical companies was 92. 7% and 97. 4%, respectively, that of acyclovir calculated by an external standard method was 0. 5% and 0. 4%, respectively, and that of D-valacyclovir calculated by a self-control method was 0. 9%. Conclusion:The method can effectively separate valacyclovir and D-valacyclovir, which is simple, accurate and reliable, and suitable for the quantity control of valacyclovir hydrochloride.
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Objective:To determinate the substances with high molecular weight in porcine anterior pituitary and adrenal cortex extracts injection. Methods:The HPLC analysis was performed on a TSK-GEL G2000SWXL(7. 8 mm × 300 mm,5 μm) column with the mobile phase of trifluoroacetic acid-acetonitrile-water(0. 05:35:65). The flow rate was 0. 5 ml·min-1, and the detection wave-length was 214 nm. Results:The substances with high molecular weight in porcine anterior pituitary and adrenal cortex extracts injec-tion was below 1%. Conclusion:The method is simple and repeatable, which can be used for the control of the substances with high molecular weight in porcine anterior pituitary and adrenal cortex extracts injection.
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Objective To investigate the epidermal growth factor receptor (EGFR),KRAS and BRAF mutations and their correlation with clinicopathological characteristics in non-small-cell lung cancer (NSCLC).Methods The mutations of exon 18,exon 19,exon 20 and exon 21 of the EGFR,codon 12,codon 13 of the KRAS and codon 600 of the BRAF gene in 143 cases of NSCLC were detected by gene sequencing.The relationship between the mutations and clinicopathological features was analyzed by SPSS 16.0.Results EGFR mutation was detected in 57 cases (39.9 %),including 2 mutations in exon 18,25 in exon 19,3 in exon 20,24 in exon 21 and 3 multiple point mutations.KRAS mutation was found in 25 cases (17.5 %),including 23 in codon 12 and 2 in codon 13.BRAF V600E mutation was detected only in 2 cases (1.4 %).No patient harboring multiple EGFR,KRAS and BRAF mutations was found.EGFR mutation rate was related to gender,smoking history,histological types,differentiation and tumor size (P < 0.05).However,no relationship was found among lymph node metastasis,pTNM stage and EGFR mutation (P > 0.05).There was no association between KRAS mutation and clinicopathological features including gender,smoking history,histological types,differentiation,tumor size,lymph node metastasis and pTNM stage (P > 0.05).Conclusions The frequency of EGFR mutation in NSCLC is high,and usually occurs in female,non-smokers,smaller tumors,better differentiation and adenocarcinomas.The frequency of KRAS mutation is not associated with the clinicolpathological features.The frequency of BRAF mutation is very low,and EGFR,KRAS and BRAF gene mutations do not occur at the same time.These results contribute to the target therapy of NSCLC.
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Objective To determine whether the CD133+ cancer stem cells lie in circumferential resection margin of rectal cancer,and to explore the relationship between CD133+ cancer stem cells and prognosis of rectal cancer.Methods The circumferential resection margin of rectal cancer was cut,one half of the tissue was stained HE for microscope observation.Both groups were labeled CD133 by immunohistochemistry.The tissues with positive circumferential resection margin were classified as experimental group,otherwise the control group.Another half was made into single cell suspension and used to detect CD133+ cancer stem cells by flow cytometry.The relapse of patients in the experimental group was following-up.Results Two cases were positive by immunohistochemistry in the experimental group (28 cases),showed that CD133+ cancer stem cells were in circumferential resection margin,but there was not marked difference compared with the control group (72 cases were all negative) (P =0.076).CD133+ cancer stem cells can be detected by flow cytometry,and the expression was higher in experimental group than that in the control group (5.19 % and 0.56 %,Z =-7.739,P < 0.01).In experimental group,the expression of CD133+ cancer stem cells in circumferential resection margin of relapse patients was significantly higher than that in patients without relapse,the difference was statistically significant [(6.57±1.72) % and (4.77±1.33) %,t =3.038,P < 0.05].Spearman rank correlation analysis showed that the proportion of CD133+ cancer stem cells in the experimental group was positively correlated with relapse (rs =0.473,P < 0.05).Conclusion There are CD133 + cancer stem cells within circumferential resection margin of rectal cancer,and the expression of CD133+ cancer stem cells in experimental group is correlated with the postoperative recurrence.
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Objective To explore the physical/chemical properties and effect of photodynamic therapy (PDT) of iminodiacetic acid modified tetraphenylporphyrin on mice bearing H22 liver cancer.Methods The UV absorbance spectrum,singlet oxygen yield and lipid-water partition coefficient were measured by UV spectrophotometry.Mice bearing H22 liver cancer as animal model were divided into control group (no drug,no light),light treated group (no drug,only light),drug treated group (only drug,no light),experimental once group (with drug and light,once) and experimental twice group (with drug and light,twice,PDT twice).Tumor inhibition rate and index of liver,spleen,lung,thymus of the mice were calculated to evaluate the antitumor activity of iminodiacetic acid modified tetraphenyl porphyrin-PDT.Results The iminodiacetic acid modified tetraphenylporphyrin had absorption at 650 nm,while its singlet oxygen yield and lipid-water partition coefficient were 2.30 and 0.52,respectively.No antitumor effect on mice bearing H22liver cancer for light treated group and drug treated group respectively.However,significant antitumor effect (P<0.001)was found at experimental group.The maximum tumor inhibition rate could be 95.15% for PDT twice group.No significant differences were observed on weight gain and liver,lung,spleen,thymus index in each group.Conclusion Iminodiacetic acid modified tetraphenylporphyrin with high singlet oxygen yield,low toxicity and significant in vivo anti-tumor activity,is suitable to be developed as an anti-tumor drug candidate for photodynamic therapy.
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<p><b>OBJECTIVE</b>To develop an HPLC method for determination of tomatine in the dried Solanum cathayanum of China Hubei Enshi.</p><p><b>METHOD</b>The analysis was performed on a YMC-Pack ODS-AA column (4. 6 mm x 150 mm, 5 microm) eluted with acetonitrile and water in gradient mode. The concentration of acetonitrile in the mobile phase changes from 20% to 100% within 60 minutes. The detection wavelength was set 203 nm. The flow rate was 1.0 mL x min(-1) and column temperature was set at 30 degrees C.</p><p><b>RESULT</b>The linear relationship of tomatine was determined within the range from 0.1-0.6 g x L(-1) (r = 0.999 7). The average recovery as 98.93% with RSD 1.2%.</p><p><b>CONCLUSION</b>A convenient and reliable method was developed to determine the content of tomatine in the dried S. cathayanum.</p>
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Chromatography, High Pressure Liquid , Methods , Chromatography, Reverse-Phase , Methods , Solanum , Chemistry , TomatineABSTRACT
Objective To investigate the impact and relationship between family income level (FIL) and prevalence rate of metabolic syndrome (MS) of golden wedding couples in Rugao city. Methods Random sample survey of golden wedding couples about MS in Rugao city was run recently, including relevant indexes of MS, insulin resistance (IR) degree (from HOMA-IR) and family income level. 587 couples were divided into three groups according to FIL by rapid cluster analysis: 196 with low FIL as group A, 196 with middle FIL as group B and 195 with high FIL as group C. The prevalence rate of MS, abnormal rates of blood pressure(BP), blood lipids, blood glucose including fasting plasma glucose (FPG) and postprandial 2 hours plasma glucose (2 h PG), body mass index (BMI) and insulin resistance (IR) of all groups were calculated and compared. Results The prevalence rate of MS in group B (17.60%) was lower than that in group A (26.02%)and group C (27.81%)(P