Effect of Palmatine Binding to MYC G-quadruplex on Proliferation and Apoptosis of Colon Cancer Cells and Molecular Mechanism / 中国实验方剂学杂志

Objective:To explore whether palmatine interferes with the proliferation and apoptosis of colon cancer HCT116 cells by binding to G-quadruplex in the promoter region of MYC proto-oncogene and its possible molecular mechanism. Method:Fluorescence spectrum was used to analyze the binding ability of palmatine to MYC G-quadruplex. Circular dichroism analysis was conducted to confirm the effect of palmatine on the configuration of MYC G-quadruplex， followed by the prediction of their binding mode based on molecular docking and the localization analysis of palmatine in HCT116 cells under a fluorescence microscope. The effects of palmatine on MYC gene transcription and MYC protein expression were determined by real-time fluorescence quantitative polymerase chain reaction and Western blot， respectively. The effects of palmatine on the viability and apoptosis of HCT116 cells were further assayed by thiazolyl blue tetrazolium bromide （MTT） assay and flow cytometry. Result:As revealed by fluorescence spectrum and molecular docking analysis， palmatine might bind to G-quadruplex of MYC gene through stacking. Circular dichroism analysis showed that palmatine could maintain the parallel configuration of MYC<italic> </italic>G-quadruplex. It was discovered in fluorescence imaging that palmatine was distributed in the nucleus and bond to G-quadruplex of MYC gene. In addition， palmatine inhibited MYC gene transcription， MYC protein expression， as well as the viability of HCT116 cells， and promoted the apoptosis of HCT116 cells. Conclusion:Palmatine is able to bind to MYC G-quadruplex to further inhibit the expression of MYC gene and protein expression， which may be one of the molecular mechanisms of palmatine in suppressing the proliferation of colon cancer HCT116 cells and facilitating their apoptosis.

Anti-allergic rhinitis effects of caffeoylquinic acids from the fruits of Xanthium strumarium in rodent animals via alleviating allergic and inflammatory reactions

Abstract The fruits of Xanthium strumarium L., Asteraceae, have been used for various diseases in Chinese folk medicine, including allergic rhinitis, tympanitis, arthritis, ozena etc. The current study was aimed to investigate the therapeutic effects of caffeoylquinic acids from fruits of X. strumarium on allergic rhinitis in animals. The toxicity test indicated that the caffeoylquinic acids have no obvious toxicity. By using HPLC assays combined with reference standards, ten caffeoylquinic acids were identified as the predominant constituents. Anti-allergic activities of the caffeoylquinic acids were evaluated using passive cutaneous anaphylaxis test and Schultz-Dale test; dimethylbenzene induced ear edema test was performed to evaluate its anti-inflammatory effect. Then, the allergic rhinitis model in rats was established to evaluate the therapeutic effects of the caffeoylquinic acids against allergic rhinitis with the following indexes: allergic rhinitis symptom scores, serum levels of pro-inflammatory cytokines, histopathological examination, and histamine release. Our study revealed that the caffeoylquinic acids showed obvious anti-allergic and anti-inflammatory properties, and its treatments were beneficial for ameliorating the nasal symptoms, decreasing pro-inflammatory cytokines, and inhibiting the releases of histamine. Collectively, the caffeoylquinic acids might be utilized as effective and safe disease therapeutic agents for allergic rhinitis.

Application of a support vector regression on prediction of bacillary dysentery combined with meteorological and air pollutants index / 中华疾病控制杂志

Objective To explore the application of support vector regression (SVR) model combined with meteorological and air pollutants index in the prediction of the cases of bacillary dysentery in Lanzhou City, so as to provide scientific reference for the prevention and control of bacillary dysentery.Methods Time series data of the reported cases of bacillary dysentery from December 2013 to August 2016, combined with the meteorological and air pollutants data, were used as training set to fit support vector regression model. The data from September 2016 to December 2017 was used as validation set to verify the model and compare the effect in fit and prediction with different models. Results A total of 7 192 bacillary dysentery cases were reported in Lanzhou City from 2013 to 2017. The correlation coefficient of meteorological and pollution factors with the cases of bacillary dysentery was more than 0.4, except air pressure. The parameters of the fit model were selected based on the integrated data, acquiring the three parameters with the smallest test error were C=5, γ=0.02 and ε=0.000 1, respectively. The validation set was used to test the different models, which showed that the integrated data model had the best predictive accuracy and robustness . The root mean squared error (RMSE) was 0.164 7 and the mean absolute percentage error (MAPE) was 16.405%. Conclusion SVR model combined with meteorological and air pollutants index is effective in the prediction of bacterial dysentery.

Effect of Coxsackievirus A16 on primary gerbil muscle cells and its mechanism / 预防医学

Objective To explore the killing mechanism induced by Coxsackievirus A16 (CV-A16) in primary muscle cells of gerbils, and to lay the foundations for elucidation the pathogenesis of CV-A16 and the further application of gerbil model. Methods The primary muscle cell model was established by digestion of trypsase/collagenase double enzyme hydrolysis. Primary muscle cells were infected by different dose of CV-A16 and the cell viability was detected by CCK-8 assays. Chromatin condensation and break were measured by Hoechst 33258 staining. The early and last stage of apoptosis cells were measured by AnnexinV/PI double staining. Expression changes of Caspase-3, Caspase-8, JNK and NF-κB pathway proteins were detected by Western Blot. Results The cell viability were 88.95％ and 64.05％ at groups of different multiplicity of infection (MOI=0.50 and 1.00), which was significantly different from those of the negative control group. The cell viability and multiplicity of infection were negative correlation (rs=-0.857, P=0.014) . The apoptosis rates were 7.2％, 21.8％ and 50.7％ at MOI=0.01,0.10 and 1.00 groups, respectively. The apoptosis rate and MOI were positive correlation (rs=1.000, P<0.001) . When the primary cells were infected by CV-A16, cleavage of Caspase-3 and Caspase-8 were detected. Western Blot assays showed that the expression of NF-κB pathway proteins IκBα, p65 and p-p65 were reduced, which was different in enterovirus 71-infected cells. The JNK kinase was actived. Conclusion CV-A16 could induce apoptosis in primary muscle cells from gerbils.

A New Method of 3-Dimensional Localization of Intraocular Foreign Bodies Using CT Imaging: A Role of Optic Nerve / 华中科技大学学报（医学）（英德文版）

Computed tomography (CT) is considered the most sensitive method for the detection of intraocular foreign bodies (IOFBs).The purpose of this study was to evaluate a new method of 3-dimentional (3D) localization of IOFBs that takes advantage of the anatomical structure of the optic nerve and to assess the clinical outcomes using this new method.Twenty-two trauma patients with IOFBs or suspected IOFBs admitted to our hospital were scanned with multislice CT (MSCT) between July and December 2003.All scanning was performed with a 16-row spiral CT in axial plane using a sequential scanning protocol.During the scanning,the eyeball of the patient was kept stable and was not allowed to rotate internally or externally.Section collimation was set at 16 mm × 0.75 mm.Table feed was 12 mm.Reconstruction index was 0.75 mm.After scanning,the reconstructed images were loaded into a workstation to create the multiplanar reconstruction images with the aid of the 3D software.We compared the localization results with the operative findings.Multiplanar reconstruction images showed IOFBs in all 22 patients.IOFBs occurred in the eyeball of 14 patients,in the wall of the eyeball of 5 patients and in the posterior orbits of 3 patients.Different surgical procedures were designed according to the localization by this new method and all IOFBs were successfully removed.All of these foreign bodies were metallic and the localization of IOFB using MSCT was consistent with that found by operative findings.It was suggested that MSCT is a simple and effective imaging modality for the localization of IOFBs.In our study,we localized the IOFBs more quickly and accurately by taking advantage of the fixed position of the intraocular segment of the optic nerve,and determined the necessary surgical parameters.

Pancreatic Neuroendocrine Neoplasms: Correlation between MR Features and Pathological Tumor Grades / 华中科技大学学报（医学）（英德文版）

This study investigated the accuracy of MRI features in differentiating the pathological grades of pancreatic neuroendocrine neoplasms (PNENs).A total of 31 PNENs patients were retrospectively evaluated,including 19 cases in grade 1,5 in grade 2,and 7 in grade 3.Plain and contrastenhanced MRI was performed on all patients.MRI features including tumor size,margin,signal intensity,enhancement patterns,degenerative changes,duct dilatation and metastasis were analyzed.Chi square tests,Fisher's exact tests,one-way ANOVA and ROC analysis were conducted to assess the associations between MRI features and different tumor grades.It was found that patients with older age,tumors with higher TNM stage and without hormonal syndrome had higher grade of PNETs (all P＜0.05).Tumor size,shape,margin and growth pattern,tumor pattern,pancreatic and bile duct dilatation and presence of lymphatic and distant metastasis as well as MR enhancement pattern and tumor-topancreas contrast during arterial phase were the key features differentiating tumors of all grades (all P＜0.05).ROC analysis revealed that the tumor size with threshold of 2.8 cm,irregular shape,pancreatic duct dilatation and lymphadenopathy showed satisfactory sensitivity and specificity in distinguishing grade 3 from grade 1 and grade 2 tumors.Features ofperipancreatic tissue or vascular invasion,and distant metastasis showed high specificity but relatively low sensitivity.In conclusion,larger size,poorlydefined margin,heterogeneous enhanced pattern during arterial phase,duct dilatation and the presence of metastases are common features of higher grade PNENs.Plain and contrast-enhanced MRI provides the ability to differentiate tumors with different pathological grades.

The Value of Transvaginal Ultrasound in Clinical Surgical Treatment of Cesarean Scar Pregnancy / 华中科技大学学报（医学）（英德文版）

The clinical value of transvaginal ultrasound in clinical surgical treatment of cesarean scar pregnancy (CSP) was evaluated.The clinical data of 104 patients with CSP admitted at Tongji Hospital from 2013 to 2016 were collected and analyzed retrospectively,including the patients' age,gestational age,the size of gestational sac or uneven mass,the uterine scar thickness,β human chorionic gonadotropin (hCG) levels and so on.Of these 104 cases,30 cases were subjected to laparotomy,29 cases to laparoscopy,27 cases to hysteroscopy,16 cases to ultrasound-guided uterine curettage,and 2 cases to conservative treatment.The transvaginal ultrasound showed that uterine scar thickness and gestational sac or uneven mass size had significant difference (P＜0.05) among different surgical methods by comparatively analyzing the patients' data.It was suggested that transvaginal ultrasound may provide the valuable reference for choosing clinical surgical procedures for CSP.

Metabolic response and dynamic nutrition support after trauma / 中华烧伤杂志

Trauma can induce a series of dynamic metabolic responses with different characteristics in three stages, including ebb phase, flow phase, and recovery phase. Nutrition support after trauma should be dynamically adjusted according to metabolic response and its individual phase, focusing on energy balance as well as metabolic modulation, adjusting immunological response to trauma by reduction of the exaggerated cytokines production, maintaining body protein balance, and enhancing wound healing.

A cross-sectional study on the prevalence of HIV drug resistance in patients receiving antiretroviral treatment in Shenqiu county, Henan province / 中华流行病学杂志

Objective To understand the prevalence of drug resistance in AIDS patients who had been receiving HAART in a long run,in Shenqiu county,Henan province.Methods This crosssectional study included 120 HIV infected patients who began receiving ART (antiretroviral therapy) in 2003.Viral loads and CD4 +T cells counts were measured,and In-house drug resistance test was performed in VL ＞ 1000 copies/ml patients.Results 114 cases out of 120 patients had complete viral load data.Among them,33 cases having viral loads less than 50 copies/ml,and the remaining viral loads showed an average of lg (4.09 ± 1.10) copies/ml.The average of CD4+ T cell counts was (377 ±2 1 8) cells/ml,with 64 (53.3％) cases showing their CD4+ T cell counts higher than 350 cells/ml.In 67 patients,58 of them showed genotypic resistance,and 40 cases showed reverse transcriptase inhibitors (RTIs) resistance.The ratios of nucleoside reverse transcriptase inhibitors (NRTIs) and nonnucleoside reverse transcriptase inhibitors (NNRTIs) resistance were 53.4％ (31/58) and 67.2％ (39/58),respectively.There were no differences of drug resistance ratio in the three treatment programs.The highest drug resistance rates in NRTIs and NNRTIs were zidovudine,lamivudin,nevirapine.However,protease inhibitors (PIs) resistance variants were not found.Conclusion The prevalence of drug-resistant strains seemed to be high in Shenqiu country,Henan province.Long-term follow-up monitoring strategy should be developed to optimize the timely treatment programs.

Evaluation of dry eye syndrom in C57BL/6.NOD-Aec1Aec2 mice / 中华实验眼科杂志

Background C57BL/6.NOD-Aec1Aec2 mouse is considered to be an idea model for the study of the pathogenesis of Sj(o)gren syndrome,but the cause of dry eye in these mice is unclear.Objective This study was to investigate the histopathological change of the ocular surfaces of C57BL/6.NOD-Aec1Aec2 mice,and to determine whether dry eye is developed spontaneously in these mice.Methods Forty-five clean C57BL/6.NOD-Aec1 Aec2 mice were used as the experiment group and forty-five C57BL/6J mice(both male and female)were used as the control group in this study.Detection of fasting blood-glucose,Schirmer' s test Ⅰ (S Ⅰ t),lissamine green staining and scoring of the corneal and conjunctival epithelium in the mice were performed at the age of 4,8,12,16 and 20weeks.Five mice from each group were sacrificed and their corneas were obtained to measure the central corneal epithelium thickness and to count the number of conjunctival goblet cells.In addition,lymphocyte infiltration in the lacrimal gland of the mice was examined with hematoxylin-eosin staining.The uhrastructure of the corneal epithelial cells and microvilli were assessed by scanning electron microscopy.The use and care of the mice were approved by the Experimental Animal Care Committee of the Third Military Medical University.Results No sign of dry eye was seen in both the 4-week-old C57BL/6.NOD-Aec1Aec2 mice and 4-week-old C57BL/6J mice.The S Ⅰ t values in 8-week-old,12-week-old,16-week-old and 20 week-old mice from the experiment group were (2.7 ±0.9) mm,(2.5 ±0.8) mm,(1.8±0.6) mm and (1.9± 0.1) mm,respectively,showing a significant reduction in comparison with those of the control mice of the same age(all P＜0.01).The amount of lissamine green staining in the C57BL/6.NOD-Aec1Aec2 mice gradually increased with age,showing elevated scores in 12-week-old,16-week-old and 20-week-old mice in the experiment group(all P＜0.01).The central corneal epithelium thicknesses were(20.18±3.75)μm,(17.01 ±5.25) μm,(14.19±5.72) μm and(12.00±3.25) μm in the 8-week-old,12-week-old,16-week-old and 20-week-old C57BL/6.NOD-Aec1Aec2 mice,respectively,which were significantly lower than those of the C57BL/6Jmice of the same age (all P＜0.01).The numbers of conjunctival goblet cells were (8.2±2.4),(6.2±2.1),(6.1 ±2.2) and (4.1 ± 2.0) in the 8-week-old,12-week-old,16-week-old and 20-week-old C57 BL/6.NOD-Aec 1Aec2 mice,respectively,showing a gradual decrease with age and a significant decline in comparison with those of the C57BL/6Jmice of the same age(all P＜0.01).Lymphocyte infiltration in the lacrimal gland and destruction of gland ducts were seen by hematoxylin-eosin staining,and acinar abnormality aggravated with aging.Reduction of corneal epithelial cells and the number of microvilli were distinguished with aging under the scanning electron microscope.The fasting bloodglucose levels of the two groups were both less than 6.0 mmol/L,and no significant difference was found between them at any age(P=0.637,0.610,0.163,0.086,0.938).Conclusions C57BL/6.NOD-Aec1Aec2 mice develop dry eye spontaneously with aging.The course of disease and characteristics of dry eye in C57BL/6.NOD-Aec1Aec2mice is similar to human dry eye.The C57BL/6NOD-Aec1 Aec2 mouse is the perfect model to study the pathogenesis of dry eye.

Comparison of nutritional status between pancreaticojejunostomy and pancreaticogastrostomy following pancreaticoduodenectomy / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To compare the nutritional status between pancreaticojejunostomy(PJ) and pancreaticogastrostomy(PG) following pancreaticoduodenectomy.</p><p><b>METHODS</b>A retrospective clinical analysis was performed on 37 patients undergoing pancreaticoduodenectomy(PD) for duodenal carcinoma and pancreatic non-epithelial tumor with PG(n=19) and PJ(n=18) in the First Hospital of Sun Yat-sen University from April 2006 to December 2010. All the patients had a needle catheter jejunostomy inserted at the conclusion of laparotomy. Postoperative early enteral nutrition and parenteral nutrition was performed for all the patients. Nutritional status of two groups was compared in body mass index (BMI), serum nutritional parameters such as albumin, transferrin and prealbumin before surgery and on 1, 3, and 6 months postoperatively.</p><p><b>RESULTS</b>There were no significant differences between PG and PJ groups in operative time, blood loss, pancreatic fistula, perioperative death, or postoperative length of hospital stay. One month after surgery, there were no significant differences in BMI [(17.1±7.0) vs. (19.0±4.8) kg/m(2), P>0.05], albumin [(30.1±0.5) vs. (32.1±1.3) g/L, P>0.05], transferrin [(1.89±0.57) vs. (2.01±0.61) g/L, P>0.05] and prealbumin[(0.18±0.05) vs. (0.18±0.09) g/L, P>0.05]. These parameters were decreased at 1 month after surgery, and gradually recovered to baseline or higher than the preoperative levels at 6 months after surgery. However, the differences were still not statistically significant between two groups.</p><p><b>CONCLUSIONS</b>The influence of PJ and PG on the postoperative nutritional status are comparable.</p>

Genotype of hantavirus in Zhejiang province / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To identify the genotype and clades of hantavirus (HV) in Zhejiang province.</p><p><b>METHODS</b>The partial S and M segment of the HV in Zhejiang province were amplified with RT-PCR using genotype-specific primers, and then were sequenced and compared with other known hantaviruses.</p><p><b>RESULTS</b>The genotype of 11 strains were HTNV and other 7 strains were SEOV by homology and phylogenesis analysis, yet the clade distribution was significantly different among foci of Zhejiang with 5 clades of HTNV and 3 clades of SEOV. There also existed special clade of HTNV named ZNB-1, ZNB-2, A3 and of SEOV named Gou3, ZJ5. The homology of M segments of ZNB-1 and ZNB-2 with other HTNV clades were 69.7%-74.0% except Nc167, A3 with other HTNV clades were 73.6%-76.3% except B78.</p><p><b>CONCLUSION</b>Zhejiang province is co-circulating with HTN and SEO. Say the least of the clades are 5 of HTNV and 3 of SEOV and there also existed special clade of HTNV and SEOV.</p>

Study on comprehensive monitoring of mouse and effect of hemorrhagic fever with renal syndrome vaccine in high prevalence areas of natural focus infectious disease of Zhejiang province in 1994--2010 / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the comprehensive monitoring mechanism of mouse and the effect of hemorrhagic fever with renal syndrome (HFRS) vaccine in the high prevalence areas of natural focus infectious disease of Zhejiang province in 1994 - 2010.</p><p><b>METHODS</b>The night trapping method was used to monitor the population proportion, density and the rate of hantavirus (HV) carriers in mice in Xikou township Longyou county in August and September from 1994 to 2010. The healthy residents in Xikou township aged 16 to 60 years were recruited. The subjects were randomly selected as vaccination group and control group according to age, sex, occupational distribution (10 178 in intervention group and 16 159 in control group). Intervention group was given purified and inactivated vaccine from suckling mouse brain, while the control group received no intervention. The prevention effect was evaluated by protective rate of vaccine.</p><p><b>RESULTS</b>The mouse population was stable in the sixteen years and the apodemus agrarius was the main type (76.5% (564/737)). The average density of mouse was 4.73% (1170/24 727). The average rate of virus carrier of mouse was 3.87% (41/1033). In 1994 - 1995, the density of mouse was 22.82% (186/815) and the rate of virus carrier was 7.0% (10/143). In 2009 - 2010, the density of mouse decreased to 2.75% (119/4330) and the rate of virus carrier was 5.5% (13/237). The average antibody positive rate of mouse from 2005 to 2010 was 4.8% (35/728) and the rate was 4.4% (6/138), 0.0% (0/113), 11.8% (16/136), 1.0% (1/104), 3.7% (4/109) and 6.3% (8/128) in each year (P < 0.01). The protective rate of HFRS vaccine was 96.2% (1 case in intervention group and 41 cases in control group).</p><p><b>CONCLUSION</b>The density of mouse decreased significantly in Zhejiang province. The rate of virus carrier of mouse is stable. The vaccine is effective.</p>

The expression and bioactivity of S gene in Vero-E6 cells using baculovirus vector / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>The S gene of a Hanta Virus (HV) Z10 strain was cloned into a baculovirus shuttle bacmid pDual-CMV contained a CMV promoter to generated a recombinant baculovirus BAC-pDual-CMV-HVS, then the recombinant baculovirus was transfected into Vero-E6 cell. The cells with recombinant baculovirus were applied to the detection of HV antiserum.</p><p><b>METHODS</b>To generate the recombinant baculovirus BAC-pDual-CMV-HVS, the sequence of CMV promoter was obtained from the plasmid pEGFP-N1 by PCR, and subsequently cloned to the baculovirus shuttle bacmid pFastBacDUAL resulting the recombinant plasmid pDual-CMV. Then the sequence of HV-S gene was inserted to the plasmid pDual-CMV, to generate the plasmid pDual-CMV-HVS. Plasmid pDual-CMV-HVS was transformed into the DH10BAC competent cells to get the recombinant baculovirus BAC-pDual-CMV-HVS. The antigen substrate slides were made by transfecting the recombinant virus into Vero-E6 cells.</p><p><b>RESULTS</b>The plasmid pDual-CMV-HVS was verified by sequencing. The recombinant virus BAC-pDual-CMV-HVS was generated according to the protocol of the baculovirus and transfected into Vero-E6 cells. The expression of the HV-S gene was verified by positive HV antiserum.</p><p><b>CONCLUSION</b>[corrected] The recombinant virus were successfully generated and applied to prepare the antigen substrate slides. The antigen substrate slides was conveniently prepared without special equipments, and can be used to detect the antiserum of HV virus.</p>

A study of immunoassay by using quantum dots to detect Hantavirus infection / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To develop a new method to detect anti-Hantavirus IgG antibodies (HV IgG) based on quantum dots (QDs) and indirect immune technique.</p><p><b>METHODS</b>The carbodiimide crosslinking method was used to couple protein G and goat antihuman IgG on the surface of water-solubility QDs. The coverglass covered HV antigen was used as carrier, and QDs-PG-IgG conjugates was used as labeled second antibody to detect the HV-IgG in the serum samples. The detecting conditions were optimized.</p><p><b>RESULTS</b>The optimum reaction time, pH and goat antihuman IgG concentration for conjugating the QDs with goat antihuman IgG were 6.0, 2h, and 20 microg/ml, respectively. The optimum working dilution of QDs-PG-IgG conjugates was 1: 200. The detection limit of the serum samples was about 1: 1280 dilution.</p><p><b>CONCLUSION</b>The method established in this study has been demonstrated to be a specific, sensitive, rapid test for detecting HV antibodies, laying the foundation of single molecule detection. The anti-fluorescence quenching ability of this method was significant improved.</p>

Studying on the prevalence and mutation pattern of N348I which related to the resistance of HIV-1 / 中华流行病学杂志

Objective To elucidate the prevalence and the mutation pattern of N348I that related to the resistance seen in the AIDS patients, in China. Methods Partial pol gene of HIV-1 comprising of full protease (PR) and reverse transeriptase (RT) was obtained from plasma samples of therapy-failure individuals (n=614) and therapy-naive individuals (n=619) by using reverse transcription polymerase chain reaction(RT-PCR). 1233 sequences were then submitted to the HIV-1 drug resistance database of the Stanford University to analyze the prevalence and the emergence pattern of N348I. Results The prevalence of N348I was 6.5％ in the therapy-failure patients and 0.8％ in the naive individuals, respectively. The prevalence of N348I was more popular among those patients whose ART regimens containing zidovudine (AZT or ZDV) than those without AZT in regimens( 14.1％ vs. 4.7％, x2=10.21, P＜0.01 ). N3481 always emerged, and combined with others mutations among patients of ART, whose frequencies were: 85.0％ in combination with thymidine analog mutations (TAMs) and 52.5％ with M184V/I, respectively. Conclusion N348I was somehow prevalent in the therapy-failure patients when using the first-line antiretroviral drugs,and it emerged as unique patterns. This study laid the ground in improving the techaology of drug resistance genotypes detection and providing theoretical basis to study the mechanism of resistance and the law of molecular evolution.

Phase I trial of icotinib, a novel epidermal growth factor receptor tyrosine kinase inhibitor, in Chinese patients with non-small cell lung cancer / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>The preclinical experiments and studies of congener drugs show icotinib, a new epidermal growth factor receptor (EGFR) tyrosine kinase inhibitor, can specifically bind to the tyrosine kinase domain of the EGFR, block the EGFR related signal, thereby inhibit the growth of tumor cell. The objective of this study was to investigate the safety, tolerability and dose-related biologic effects of icotinib in patients with non-small cell lung cancer (NSCLC) in a Chinese patient population.</p><p><b>METHODS</b>This was an open-label, phase I, dose escalation, safety/tolerability trial of oral icotinib (100 to 400 mg), administered twice per day for 28-continuous-day cycles until disease progression or undue toxicity.</p><p><b>RESULTS</b>Forty patients with stage IIIB (15%) or IV (85%) NSCLC were included in the study. They had mainly adenocarcinoma (85%), with a performance status (PS) of 0 (45%) or 1 (55%) and less than half the patients (45%) had histories of smoking and all were pretreated by at least one regimen of chemotherapy. Patients were assigned to three dose levels of 150 mg b.i.d, 200 mg b.i.d, or 125 mg t.i.d. The follow-up periods ranged from 5 to 80 weeks. Adverse events were found in 35% patients, most of which were mild and reversible. The adverse events mainly occurred in the first 4 weeks and included rash (25%), diarrhea, nausea and abdominal distention. One definite interstitial lung disease (ILD) was found in a patient in the dose of 200 mg b.i.d. According to an 8-week assessment, one (2.5%) patient receiving 150 mg gained complete response (CR) that persisted for 44 weeks, seven (17.50%) patients had partial remission (PR), and 18 (45%) patients had stable disease (SD). The objective response including CR + PR was 20%. The median time of progression-free survival for the 40 patients was 20 weeks (range: 12 to 32 weeks). The response was not affected by pathological type, history of smoking, or numbers of previous therapeutic regimens. No relationship between dose, response, adverse effect, or duration of the study was observed.</p><p><b>CONCLUSIONS</b>Icotinib, given as oral twice daily, showed favorable safety and tolerability. Mild and reversible rash, diarrhea, and nausea were the main adverse events. Antitumor activity was obvious at each dose in heavily pretreated patients. Pharmacodynamic evaluations and further phase II/III trials are in progress.</p>

Screening program on novel drug resistance mutations of subtype B' in human immunodeficiency virus type 1 in China / 中华流行病学杂志

Objective To screen the level of novel drug resistance mutations in subtype B' in China. Methods 451 pol sequences collected from the previous study, which including 354 AIDS patients who had received antiretroviral treatment(ART)and 97 the untreated patients. Entire protease gene(codous 1-99)and full-length reverse transcriptase gene(codons 1-560)were included.Variation of mutations between the treated and the untreated patients with consensus/ancestral sequences were compared and the mutations with higher frequencies in the treated patients than in the untreated patients were screened before submitting the mutations to the Stanford HIV Drug Resistance Database(SHDB)(http://hivdb.stanford.edu/). Relation between the mutations and resistance preliminarily was then analyzed, according to the information including SHDB. Results Frequencies of 7 mutations at 6 positions, DI23E, V292I, K366R, T369A, T369V, A371V and 1375V, 2 at DNA polymerase domain and 5 at connection domain of reverse transcriptase(RT)were higher in the treated patients than in the untreated patients. The information of 7 mutations including the SHDB showed that 7 mutations were major variants at corresponding positions, and theirs frequencies were higher in the treated patients using some drugs, than in the untreated patients. Conclusion 7mutations being screened from the China subtype B were possibly associated with the resistance,which called for the construction of mutated viruses by site-directed mutagenesis to identify their effects on the susceptivity of different drugs.

Pharmacokinetics comparison of amphotericin B using three administrating approach in a rabbit model / 中华实验眼科杂志

Background Intracameral or intracorneal administration of amphotericin B (AMB) can achieve significant therapeutic efficacy in the treatment of recalcitrant fungal keratitis in cases that do not respond to conventional antifungal therapy. However, the ocular pharmacokinetics of the two routes of administration is unclear.Objective The goal of this study was to investigate the level of amphotericin B in cornea and aqueous humor of rabbits after administration of AMB via three different routes. Methods Forty-five healthy domestic rabbits were randomly divided into three groups. 1% amphotericin B of 10 μg was intrastromally or intracamerally injected into 15 rabbits, respectively,in group A and group B. Topical 0. 25% amphotericin B was topically administered to the eyes with corneal epithelial debridement (group C). Experimental animals were sacrificed and the corneas and aqueous humor samples were obtained for the detection of levels of amphotericin B at 30 minutes,6 hours, 1 day,3 and 7 days by high-performance liquid chromatography (HPLC). Results Calibration curves were linear over the range of 0. 10-100. 00 mg/L. The concentration of 0. 10 mg/L was the lowest quantifiable limit. The recovery of amphotericin B ranged from 89. 1% -95.7% from aqueous humor samples and 81.4% -83.6% from the cornea samples. After a single injection,effective drug levels were achieved and maintained for 7 days in cornea in group A, exceeding the minimum inhibitory concentration at which 90% of isolates are inhibited (MIC90) for a wide spectrum of fungi and molds with significant differences in comparison with group B and group C ( P＜0. 05 ). Effective drug levels were achieved in the aqueous humor in group B at 30 minutes after a single injection, but drug levels decreased dramatically within 6 hours. The evident differences were found between group B and group A or group C (P＜ 0.05). A considerable amount of amphotericin B was detected in the cornea and aqueous humor in group C within 1 day.Conclusion Effective high drug levels can be reached in rabbit cornea and aqueous humor after intrastromal and intracameral injection, respectively. Penetration of topical amphotericin B was greatly elevated after epithelial debridement.

Allele-specific real-time PCR testing for minor HIV-1 drug resistance mutations: assay preparation and application to reveal dynamic of mutations in vivo / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>It is very important for the clinical management to test for minor HIV-1 resistance mutations accurately and sensitively. The conventional genotypic assays of HIV drug resistance detection based on sequencing can only discriminate the mutations which present in more than 20% - 30%. The aim of this study was to evaluate allele-specific real-time PCR (ASPCR) to detect the resistance-related mutations located at positions 103, 184 and 215.</p><p><b>METHODS</b>We developed the allele-specific PCR assay, using the most common drug resistance mutations in Chinese AIDS patients, K103N, M184V/I, T215F/Y as a model system. The standards were constructed by cloning the wild-type and mutant DNA fragments into the T-vector. We designed specific primers to discriminate mutant templates in the real-time PCR using SYBR green as a fluorescence reporter. And then we evaluated the ASPCR assay and tested 140 clinical samples using this method.</p><p><b>RESULTS</b>The sensitivities of ASPCR assay were 0.04% for K103N, 0.30% for M184I, 0.40% for M184V, 0.03% for T215F and 0.02% for T215Y. The intra-assay and inter-assay coefficients of variation were less than 0.42. One hundred and forty plasma samples were tested by ASPCR and dynamic resistance curves of ten patients were obtained.</p><p><b>CONCLUSIONS</b>Drug resistance emerged half a year after the start of antiretroviral therapy. The mutation of T215Y emerged 1 to 1.5 years after starting treatment and then increased rapidly. The ASPCR assay we developed was a sensitive, accurate and rapid method to detect the minor HIV-1 variants and it can provide earlier and more drug-resistance information for HIV research and AIDS antiretroviral therapy.</p>