Application of high-speed counter current chromatography in extraction and separation of alkaloids in natural products / 中国中药杂志

Alkaloids, widespread in plants, have a series of pharmacological activities and have been widely used to treat various diseases. Because alkaloids are usually presented in multicomponent mixtures and are deeply low in content, they are very difficult to extract and separate by traditional methods. High-speed counter current chromatography(HSCCC) is a kind of liquid-liquid chromatography without solid support phase, which has the advantages of large injection volume, low cost, and no irreversible adsorption. Compared with the traditional methods of extraction and separation of alkaloids, HSCCC can ensure the separation of many different alkaloids at one time, with a high recovery and large amount. In this paper, the advantages and disadvantages of HSCCC compared with traditional separation methods were discussed and the solvent system and elution mode of HSCCC used to separate alkaloids in recent years were summarized by referring to the relevant literature to provide some references for the separation of alkaloids by HSCCC.

Alterations of macular ganglion cell layer thickness and optic disc parameters in patients with early diabetic retinopathy / 国际眼科杂志(Guoji Yanke Zazhi)

AIM: To compare the changes of optic disc parameters, peripapillary retinal nerve fibers layer(pRNFL)thickness and macular ganglion cell layer(mGCL)thickness among patients with early diabetes retinopathy and healthy controls by Cirrus HD-optical coherence tomography(OCT).METHODS: In this cross-sectional comparative study, 45 non-diabetic retinopathy(NDR), 52 mild nonproliferative diabetic retinopathy(NPDR), 55 moderate NPDR with type 2 diabetes mellitus(T2DM)and 64 age-matched healthy controls were included. The fasting blood glucose(FBG), duration of diabetes, glycosylated hemoglobin(HbA1c)and past history of the patients were collected in detail. Optic disc parameters(i.e., binocular RNFL thickness symmetry percentage, rim area, optic disc area, cup-to-disc ratio, cup volume), pRNFL thickness and mGCL thickness were measured by Cirrus HD-OCT. The comparison of different groups was performed by one-way analysis of variance.RESULTS: Compared with the control group, the binocular RNFL thickness symmetry percentage and rim area were significantly decreased, while the average C/D and vertical C/D were significantly increased in the NDR group, mild NPDR group and moderate NPDR group(all P<0.05). Compared with the control group, the peripapillary RNFL thicknesses(superior, temporal, inferior, nasal)and macular GCL thickness(average, minimum, superior, supero-temporal, infero-temporal, inferior, supero-nasal, and infero-nasal)became thinner in the NDR group, mild NPDR group, and moderate NPDR group(all P<0.05).CONCLUSION: Patients with early DR have significantly decreased binocular RNFL thickness asymmetry, rim area, pRNFL and mGCL thickness, while they have significantly increased cup-to-disc ratio when compared to healthy controls. The results support the statement that DM causes inner retinal neurodegenerative changes even in T2DM patients without overt microangiopathy.

Clinical application of botulinum toxin type A in the treatment of thyroid-associated ophthalmopathy / 国际眼科杂志(Guoji Yanke Zazhi)

Thyroid-associated ophthalmopathy(TAO)is an autoimmune inflammatory disease involving multiple orbital tissues with a variety of clinical manifestations, which has serious effects on the life quality of patients.Interventions of TAO mainly include medical treatment to stabilize thyroid function, reduce inflammation and regulate immune function, as well as surgical treatment to relieve ocular symptoms. Botulinum toxin type A can paralyze muscles by blocking nerve impulse conduction at the neuromuscular junction, which is of certain therapeutic value for restrictive strabismus due to extraocular muscle involvement and upper eyelid retraction due to involvements of levator palpebrae superioris and Müller's muscle in TAO patients, especially when they have surgical contraindications, lack surgical opportunity, or refuse surgery. This paper reviews the application of botulinum toxin type A in the treatment of TAO, focusing on its pharmacological mechanism, dosage, effectiveness, and possible complications when treating restrictive strabismus and upper eyelid retraction, and discussing potential therapeutic values of botulinum toxin type A for intraocular pressure elevation, glabellar frown lines and dry eye caused by extraocular muscle compression in TAO patients, in order to provide a reference for clinical intervention.

Research progress on chemical constituents and pharmacological activities of iridoid glycosides in Lonicera japonica / 中国中药杂志

As a traditional Chinese medicinal material, Lonicera japonica has a long medicinal history. The chemical constituents of Lonicera japonica are complex, mainly including iridoid glycosides, flavonoids, triterpenes, organic acids and volatile oil. Iridoid glycosides account for a higher proportion. In addition, modern pharmacological studies have shown that the iridoid glycosides have many pharmacological activities such as antivirus, anti-inflammation, anti-tumor, liver protection and lowering blood sugar. This review intends to systematically summarize the iridoid glycosides identified from Lonicera japonica and their pharmacological activities by searc-hing Chinese and English databases, in order to provide a reference for the further development and utilization of Lonicera japonica and for the improvement of quality standards of medicinal materials.

Three new secoiridoid glycosides from the flower buds of Lonicera japonica / 中国天然药物

Three new secoiridoid glycosides, named lonijapoglycol A (1), aldosecolohanin C (2) and aldosecolohanin B (3), together with three known ones (4-6), have been isolated from the flower the buds of Lonicera japonica. All the structures were identified by spectroscopic analyses. Lonijapoglycol A (1) expressed significant anti-inflammatory activity to inhibit the release of β-glu-curonidase induced by platelet-activating factor in rat polymorphonuclear leukocytes with an IC value of 3.76 μmol·L.

Effect of Shen-Fu Injection () on Hemodynamics in Early Volume Resuscitation Treated Septic Shock Patients / 中国结合医学杂志

OBJECTIVE@#To investigate the hemodynamic effect of Shen-Fu Injection (, SFI) in early volume resuscitation treated septic shock patients by monitoring pulse indicator continuous cardiac output (PICCO).@*METHODS@#All septic shock patients admitted in the Intensive Care Unit of the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from January 1st, 2014 to December 31th, 2015, were reviewed, and totally 65 were enrolled in this study. They were assigned to SFI group (33 cases) and control group (32 cases). All 65 patients underwent conventional treatment mainly including volume resuscitation, antibiotics and vasoactive drugs therapy. The patients of the SFI group received additional 100 mL of SFI intravenously every 12 h. In all 65 patients, the PICCO arterial catheter and vein catheter were implanted within 1 h after the diagnosis of septic shock. In the course of early volume resuscitation, hemodynamic data of patients were recorded by PICCO monitor at 0, 12, and 24 h after the catheter implantation.@*RESULTS@#The hemodynamic indices of the two groups showed no significant differences at the beginning of 0 h (P>0.05). At 12 and 24 h, the hemodynamic indices of SFI group were significantly improved in comparison with the control group (P0.05).@*CONCLUSION@#SFI significantly improved hemodynamic indices such as CI, GEDI, MAP and HR in early volume resuscitation treated septic shock patients.

Clinical Efficacy of Low-dose Decitabine Combined with CAG Regimen in Patients with Myelodysplastic Syndrome-refractory Anemia with Excess Blasts(MDS-RAEB) / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the clinical efficacy of low-dose decitabine combined with CAG regimen in patients with myelodysplastic syndrome-refractory anemia with excess blasts (MDS-RAEB) through retrospective analysis.</p><p><b>METHODS</b>Thirty-six patients with MDS-RAEB who ever received low-dose decitabine combined with CAG regimen were enrolled into decitabine + CAG group and 40 patients with MDS-RAEB treated by decitabine alone in our center were enolled into the control group. The clinical characteristics, efficacy and adverse reactions (AE) were compared between the 2 groups.</p><p><b>RESULTS</b>Compared with the control group, the overall response rate (ORR) [complete remission (CR)+partial remission (PR) + hematologic improvement (HI) rate] of the decitabine+CAG group was higher (83.3% vs 62.5%)(P=0.043), and the AEs were not significantly increased. Cytopenia grade ≥3 and infection after treatment were the most prevalent AEs, which occurred in the early stage (within the first 2 cycles) and gradually decreased later. Other non-hematologic AEs were infrequent.</p><p><b>CONCLUSION</b>Low-dose decitabine combined with CAG regimen has better clinical efficacy for patients with MDS-RAEB than that of decitabine alone. It is worthy to be applied in clinic, especially for these patients who are not ineligible for hematopoietic stem cell transplantation.</p>

Inhibitory effect of ischemic postconditioning on autophagy induced by fo-cal cerebral ischemia reperfusion in rats / 中国病理生理杂志

AIM:To investigate the effect of ischemic postconditioning ( IPC) on autophagy induced by focal cerebral ischemia reperfusion ( I/R) in rats.METHODS:Healthy male SD rats were assigned randomly into sham-opera-tion (sham) group, I/R group and IPC group with 10 rats in each group.The rats in sham group were only exposed the right common , internal and external carotid artery surgically .The rats in I/R group were subjected to right middle cerebral artery occlusion (MCAO) by the modified Longa suture method for 2 h followed by 24 h of reperfusion.The rats in IPC group were subjected to MCAO for 2 h followed by reperfusion of the ipsilateral common carotid artery occlusion for 10 s for 5 episodes, and then reperfusion for 24 h.Autophagy was obeserved by transmission electron microscopy (TEM).The pro-tein levels of mammalian target of rapamycin (mTOR), p-mTOR and microtubule associated protein light chain 3 (LC3)-II in brain tissue of the rats were determined by Western blot .Pathological changes of brain tissue were observed by HE staining.RESULTS:The protein levels of mTOR and p-mTOR in IPC group were significantly higher than those in I/R group (P<0.05).The expression of LC3-II in IPC group was significantly lower than that in I/R group (P<0.01).The cerebral infarction area and brain water content in IPC group were significantly lower than those in I /R group (P<0.01). HE staining showed that neurons degeneration and necrosis in IPC group were significantly alleviated compared with I /R group.TEM observation showed that IPC revealed fewer autophagosomes , with much less severe cell damage than that in I/R group.CONCLUSION:IPC reduces brain ischemia reperfusion damage by decreasing autophagy of brain cells , which might be related to the activation of mTOR .

Campylobacter jejuni AhpC protein B cell antigen epitope prediction and antigenic analysis / 中国人兽共患病学报

To provide evidence for Campylobacter jejuni (C.jejuni) vaccine research,the B cell epitope of AhpC protein were predicted,and the antigenicity was analyzed.AhpC was found locating in outer membrane of C.jejuni without transmem brane structure and no signal peptide by bioinformatics software TMHMM Server V2.0,SignalP 4.1.There were seven B-cell linear epitopes in AhpC predicted by IEDB.Then,the AhpC protein and chemically synthesized antigenic epitopes of C.jejuni were used as antigens,and the AhpC antibody of C.jejuni was used as the primary antibody,the antigens of predominant linear B cell epitopes were detected by ELISA and dot blot.Results showed that one epitope of B cell epitopes (AhpC4-16) was able to recognize the antibodies of C.jejuni AhpC and had strong antigenicity,indicating that could be used in the follow-up vaccine research.

Characteristics of drug resistance and molecular typing for Salmonella in diarrhea patients from four hospitals in Shenzhen / 中国人兽共患病学报

We analyzed the antimicrobial resistance and pulsed field gel electrophoresis (PFGE) patterns of Salmonella strains of infectious diarrhea isolated four hospitals in Shenzhen during 2012-2016.A total of 137 Salmonella strains collected in 4 hospitals in Shenzhen were identified by serotyping,micro-dilution method was used to detect the antimicrobial susceptibility of the isolates.Pulsed field gel-electrophoresis(PFGE) was used for molecular typing and cluster analysis of S.typhimurium and S.enterica strains.Results showed that 137 strains of Salmonella were divided into 30 kinds of serotype.S.typhimurium was the predominant serotype.The drug susceptibility test indicated that the antibiotic resistance of the strains to chloromycetin,ampicillin and amoxicillin were more serious than other drugs,the resistance to penicillins,cephalosporins,aminolycosides and macrolides showed increase trends year by year.Of 49 strains of S.typhimurium were divided 38 molecular patterns by PFGE,with similarity ranged from 65.5％ 100％,TY18 was the main PFGE pattern.The 27 strains of S.enterica were divided into 7 molecular patterns by PFGE,with similarity ranged from 61.8％-100％,EN7 was the main PFGE pattern.The status of drug resistance of infectious diarrhea isolates was rather severe,PFGE patterns showed diversity,suggesting the diarrhea cases were distributed sporadically.Partial PFGE patterns and its corresponding antidrug spectrum have certain aggregation relationship.

Inhibitory effect of ischemic postconditioning on autophagy induced by fo-cal cerebral ischemia reperfusion in rats / 中国病理生理杂志

AIM:To investigate the effect of ischemic postconditioning ( IPC) on autophagy induced by focal cerebral ischemia reperfusion ( I/R) in rats.METHODS:Healthy male SD rats were assigned randomly into sham-opera-tion (sham) group, I/R group and IPC group with 10 rats in each group.The rats in sham group were only exposed the right common , internal and external carotid artery surgically .The rats in I/R group were subjected to right middle cerebral artery occlusion (MCAO) by the modified Longa suture method for 2 h followed by 24 h of reperfusion.The rats in IPC group were subjected to MCAO for 2 h followed by reperfusion of the ipsilateral common carotid artery occlusion for 10 s for 5 episodes, and then reperfusion for 24 h.Autophagy was obeserved by transmission electron microscopy (TEM).The pro-tein levels of mammalian target of rapamycin (mTOR), p-mTOR and microtubule associated protein light chain 3 (LC3)-II in brain tissue of the rats were determined by Western blot .Pathological changes of brain tissue were observed by HE staining.RESULTS:The protein levels of mTOR and p-mTOR in IPC group were significantly higher than those in I/R group (P<0.05).The expression of LC3-II in IPC group was significantly lower than that in I/R group (P<0.01).The cerebral infarction area and brain water content in IPC group were significantly lower than those in I /R group (P<0.01). HE staining showed that neurons degeneration and necrosis in IPC group were significantly alleviated compared with I /R group.TEM observation showed that IPC revealed fewer autophagosomes , with much less severe cell damage than that in I/R group.CONCLUSION:IPC reduces brain ischemia reperfusion damage by decreasing autophagy of brain cells , which might be related to the activation of mTOR .

Biomechanical properties of trabecular bones in femoral head under loads with different angles / 医用生物力学

Objective To investigate the effects from loads with different angles on morphological and biomechanical properties of trabecular bones in femoral head, so as to provide theoretical basis for studying biomechanical mechanism of necrosis and collapse of femoral head. Methods Ninety-four specimens of 12-month-old ovine trabecular bones in femoral head and forty-three specimens of human cadaver trabecular bones in femoral head were prepared. According to different angles between loading direction and principle compression direction, all the trabecular bones were divided into five groups by 10° interval (i.e. varus 10° and 0°, valgus 10°, 20° and 30°) to simulate the reduction condition under different Garden index after internal fixation of femoral neck fractures. Micro-CT scanning and calculation, compression failure test on ovine trabecular bones in femoral head and cyclic compression test on human cadaver trabecular bones in femoral head were performed to investigate morphological and mechanical indices, including BV/TV (bone volume vs. total volume), BS/BV (bone surface vs. bone volume), Tb.Th (thickness of trabecular bone), Tb.N (number of trabecular bone), Tb.Sp (trabecular separation), elastic modulus, ultimate strength, yield strength, initial secant modulus and number of cycles. Results When the angle between loading direction and principle compression direction of trabecular bones was 0°, BV/TV, Tb.Th, elastic modulus, ultimate strength, yield strength, initial secant modulus and number of cycles for trabecular bones were the maximum while BS/BV and Tb.N were the minimum, and all the formers presented decreasing tendency while BS/BV and Tb.N showed increasing tendency along with the angle increasing. ConclusionsAlong with the angle changes, the tendency of BV/TV and ultimate strength for 12-month-old ovine trabecular bones in femoral head displayed as the same as human trabecular bones in femoral head. Both the morphological and biomechanical properties of trabecular bones in femoral head will decrease when the angle between loading direction and principle compression direction of trabecular bones increases. The more the Garden index deviating from 160°, the more likely trabecular bones in femoral head to be damaged.

The selective dilatation effects of iptakalim on pulmonary arterioles in hypoxic pulmonary hypertensive rat / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To study the selective dilatation effects of iptakalim (Ipt), a novel ATP-sensitive potassium channel opener, on pulmonary arterioles in hypoxic pulmonary hypertensive rat.</p><p><b>METHODS</b>SD male rats were divided into 3 groups, control group, the rest were fed in hypoxic and normobaric environment (O2 10% +/- 0.5%, 8 h/d and 6 d/week) and divided into hypoxia group and hypoxia plus acetazolamide (Acz) group (hypoxic rats were treated with ig acetazolamide (Acz) 80 mg x kg(-1) d(-1)) . After 12 weeks, pulmonary arteriole rings about (197 +/- 4) microm were isolated and the tension of hypoxic pulmonary arterioles pre-contracted by 6 nmol/L endothelin-1 (FT-1) was observed with wire myograph system model (DMT 610 m). The relaxing response of hypoxic pulmonary arterioles induced by different concentration of Ipt were detected and endothelial activity was also tested by acetylcholine.</p><p><b>RESULTS</b>10(-5) mol/L acetylcholine (ACh)-mediated vasodilatation was greatly reduced in the hypoxic group than those in control group (P < 0.01) and there was no significant difference between Acz treatment group and control group (P > 0.05). Ipt at the concentration ranging from 10(-11) mol/L to 10(-4) mol/L, caused dose dependent vasodilation on both hypoxic pulmonary arterioles and Acz treatment group (P > 0.05), but not on normal group.</p><p><b>CONCLUSION</b>The endothelial function of pulmonary arterioles was damaged under hypoxic pulmonary hypertensive state, and Ipt showed selective dilatation effects on hypoxic pulmonary arterioles. Acz could improve the dysfunction of endothelial cells induced by hypoxic pulmonary hypertensive state, which didn't affect the selective dilatation effects of Ipt on hypoxic pulmonary arterioles.</p>

Application of combined telomerase activity analysis and immunocytochemistry in cytopathologic diagnosis / 中华病理学杂志

<p><b>OBJECTIVE</b>To evaluate the application of traditional cytomorphology, telomerase activity analysis and immunocytochemistry in cytopathologic diagnosis of pleural effusion and bronchoalveolar lavage samples.</p><p><b>METHODS</b>A total of 123 agar-paraffin double-embedded pleural effusion and bronchoalveolar lavage fluid samples were enrolled into study. The cytomorphologic features were reviewed and correlated with immunocytochemical findings and telomerase activity.</p><p><b>RESULTS</b>Telomerase activity was detected in 53 specimens using the real-time telomeric repeat amplification protocol. Amongst the cases studied, 39 samples (31.7%) contained overtly malignant cells while 20 cases (16.0%) were equivocal by conventional cytology. After verification by immunocytochemistry and clinical follow-up data, the diagnostic accuracy of telomerase activity and cytology was 87.0% and 82.1%, respectively. The sensitivity (97.6%) and specificity (100.0%) of cytology examination, when combined with telomerase activity analysis, were greater than those of cytology examination or telomerase activity analysis alone.</p><p><b>CONCLUSIONS</b>Telomerase activity analysis can be used as an adjunctive investigative tool in cytology assessment of pleural effusion and bronchoalveolar lavage samples. The diagnostic accuracy can be further improved with the application of immunocytochemistry on agar-paraffin double-embedded cell block tissues.</p>

Formaldehyde inhalation may damage olfactory bulb and hippocampus in rats / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of formaldehyde inhalation on the morphological damage, and Glu, GABA and NOS contents in olfactory bulb and hippocampus of rats.</p><p><b>METHODS</b>Twenty SD rats were equally divided into two groups: rats in the control group inhaled fresh air, while the animals in experimental group were exposed to the air containing formaldehyde (12.5 mg/m(3), 4 h/d) for 7 days. Then rats were sacrificed and frozen sections of olfactory bulb and hippocampus were prepared. The morphological changes were examined and the Glu, GABA and NOS contents were detected using Nissl-staining, immunohistochemistry and Western blot, respectively.</p><p><b>RESULT</b>Compared with the control group, there was a significant confusion and shrink of neuron morphology in experimental group, the number and staining intensity of Glu and NOS positive cells and protein contents were reduced. The protein expression of GABA was also decreased in the formaldehyde group.</p><p><b>CONCLUSION</b>Formaldehyde inhalation can cause a severe morphological damage of olfactory bulb and hippocampus in SD rats,which may further impair memory and learning ability through the reduction of Glu, GABA and NOS expression.</p>

Changes of plasma cytokines in patients with severe hepatitis treated with a probiotic preparation / 中华肝脏病杂志

<p><b>OBJECTIVE</b>To study the changes of plasma cytokines in patients with severe hepatitis treated with a probiotic preparation.</p><p><b>METHODS</b>One hundred twelve patients with severe hepatitis treated with a basic regimen were randomly divided into a probiotic preparation treatment group and a control group. Their plasma cytokines such as tumor necrosis factor alpha, interleukin-2, interleukin-6, interleukin-10, were determined by conventional techniques.</p><p><b>RESULTS</b>Clinical symptoms of 84.5% of the treatment group were obviously improved. The treatment effectiveness in this group was better than that in the control group (X2=8.888). It showed that liver functions were significantly improved compared to those of the control group . After the probiotic preparation therapy, there were significant decreases in the concentrations of TNFa and IL-6. TNFa and IL-6 in the treatment group and in the control group were (109.4+/-14.7) pg/ml vs (128.7+/-18.8) pg/ml and (84.3+/-20.1) pg/ml vs (109.1+/-18.7) pg/ml respectively. However, the concentrations of IL-10 and IL-2 in the treatment group increased obviously: IL-2 was (59.8+/-12.2) pg/ml vs (47.1+/-6.7) pg/ml and IL-10 was (30.6+/-6.6) pg/ml vs (22.5+/-6.1) pg/ml.</p><p><b>CONCLUSION</b>The probiotic preparation treatment effectively reduces the TNFa and IL-6 and increases the concentrations of IL-10 and IL-2 in the blood of the patients with severe hepatitis.</p>

BRCA1/2 associated hereditary breast cancer / 浙江大学学报（英文版）（B辑：生物医学和生物技术）

Breast cancer is one of the leading causes of death in women today. Some of the patients are hereditary, with a large proportion characterized by mutation in BRCA1 and/or BRCA2 genes. In this review, we provide an overview of these two genes, focusing on their relationship with hereditary breast cancers. BRCA1/2 associated hereditary breast cancers have unique features that differ from the general breast cancers, including alterations in cellular molecules, pathological bases, biological behavior, and a different prevention strategy. But the outcome of BRCA1/2 associated hereditary breast cancers still remains controversial; further studies are needed to elucidate the nature of BRCA1/2 associated hereditary breast cancers.

Estrogen receptor alpha variant ERalpha46 mediates growth inhibition and apoptosis of human HT-29 colon adenocarcinoma cells in the presence of 17beta-oestradiol / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Estrogen is involved in suppression of colon cancer development and exerts its function via estrogen receptors alpha and beta (ERalpha, ERbeta). The recently identified ERalpha46 resulted from exon 1-deletion from the 66-kDa full length form of ERalpha66 is devoid of the transactivation domain AF-1, whose function remains largely unknown.</p><p><b>METHODS</b>In this study, we compared the expression of ERalpha46 mRNA in 32 normal colorectal tissues and their matched colorectal cancer tissues by real-time quantitative polymerase chain reaction (PCR). Human colon adenocarcinoma cell HT-29, that has low endogenous expression of ERalpha46, was transfected with ERalpha46-expression vector; methyl thiazolyl tetrazolium (MTT) assay, flow cytometry, DNA fragmentation and TUNEL staining were used to evaluate the proliferation and apoptosis status of the cells in the presence of 17beta-oestradiol.</p><p><b>RESULTS</b>Higher ERalpha46 mRNA levels were observed in normal colorectal tissues than in the corresponding cancer tissues. ERalpha46-transfected cells showed a significantly decreased growth rate than control cells and an accumulation of cells in the G(0/1) phase and a reduced proportion of cells in G(2)/M phase after exposed to 10(-8) mol/L 17beta-oestradiol. There were also more positive TUNEL stained cells in ERalpha46-transfected cells than the control cells in the presence of 17beta-oestradiol (P < 0.05).</p><p><b>CONCLUSIONS</b>These data suggest that ERalpha46 may be involved in the development and/or progression of colorectal cancer via mediating growth inhibition and apoptosis of cancer cells in the presence of 17beta-oestradiol.</p>

Status and clinicopathologic implication of epidermal growth factor receptor mutation in non-small cell carcinoma of lung / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate mutations of epidermal growth factor receptor (EGFR) exon 19 and 21 in non-small cell lung carcinoma and to explore their clinicopathological correlations.</p><p><b>METHOD</b>DNA was extracted from the excised tumor specimens of 66 non-small cell lung carcinoma patients by traditional phenol-chloroform and ethanol precipitation. Exons 19 and 21 were amplified by polymerase chain reaction (PCR), followed by direct sequencing in both sense and antisense directions.</p><p><b>RESULTS</b>EGFR somatic mutations were present in 11 of 66 patients (16.7%), including 7 cases of in-frame deletion involving exon 19 and 4 cases of amino acid substitution involving exon 21. Mutations were more frequently observed in women (9/34, 26.5%) than in men (2/32, 6.3%), in adenocarcinomas (10/43, 23.3%) than squamous (0/13) and adenosquamous carcinomas (1/10). There was no difference in the mutation rates between smokers and non-smokers. Those with adenocarcinoma with bronchiolo-alveolar carcinoma (BAC) components had higher frequency of EGFR mutation (6/11) than those without non-BAC element (4/32, 12.5%).</p><p><b>CONCLUSIONS</b>The mutations appear to occur in highly selected subgroups of lung cancer patients: adenocarcinomas with BAC components and patients of the female gender. The results may offer practical approach to the rapid identification of lung cancer patients who likely respond to EGFR inhibitor therapy.</p>

P2Y purinergic receptor activated PI-3K/Akt signaling pathway in regulation of growth and invasion of prostatic cancer / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate P2Y purinergic receptor activated PI-3K/Akt signaling pathway in the regulation of growth and invasion of prostate cancer in vitro.</p><p><b>METHODS</b>Western blot was used to detect phosphorylation of Akt (a downstream target molecule of PI-3K) by P2Y receptor agonist in 1E8 cells (a highly metastatic subclone derived from PC-3 prostatic cancer cell line). Cell counts, flow cytometry, Matrigel invasion assay, wound healing assay and gelatin zymography were used to detect changes of biological behaviors of 1E8 cells after P2Y receptor activation.</p><p><b>RESULTS</b>AMP-PNP, one non-hydrolysis ATP analogue and P2Y receptor agonist, induced significant phosphorylation of Akt in a time- and dose-dependent manner in IE8 cells. LY294002, a specific inhibitor of PI-3K, effectively blocked Akt phosphorylation induced by AMP-PNP. Continuous exposure to AMP-PNP induced significant growth inhibition of 1E8 cells (inhibition rate at 50.2% at the 8th day), and this inhibition was mainly due to an arrest at S phase of the cell cycle (the S phase fraction of AMP-PNP treated cells was 22.3% higher than that of the control). Application of LY294002 did not reverse the growth inhibition effect of AMP-PNP. Matrigel invasion assay showed that AMP-PNP stimulation increased invasive ability of 1E8 cells, and this effect was effectively blocked by LY294002. No significant changes in the activation of MMP-2 and MMP-9 were detected by gelatin zymography, although wound healing assay showed 21.2% increase in cell migration after AMP-PNP treatment.</p><p><b>CONCLUSIONS</b>PI-3K/Akt signaling pathway participates in P2Y receptor-stimulated prostate cancer invasion by enhancing cell motility, rather than up-regulating MMP-2 and MMP-9 activities. PI-3K signaling pathway plays an important role in prostate cancer proliferation, but is not involved in P2Y receptor mediated growth inhibition.</p>