LncRNA DRAIC regulates the proliferation, apoptosis, migration and invasion of lung adenocarcinoma cells by targeting let-7i-5p / 中华肿瘤杂志

Objective: To investigate the effects of lncRNA DRAIC on proliferation, apoptosis, migration and invasion of lung adenocarcinoma cells and its mechanism. Methods: Reverse transcription-quantitative real-time polymerase chain reaction (RT-qPCR) was used to detect the expression of DRAIC in lung cancer tissues and corresponding adjacent normal tissues of 40 patients with lung adenocarcinoma who underwent surgery in Tangshan People's Hospital from 2019 to 2020. Lung adenocarcinoma cells A549 and H1299 were cultured in vitro and divided into si-NC group, si-DRAIC group, miR-NC group, let-7i-5p mimics group, si-DRAIC+ inhibitor-NC group, and si-DRAIC+ let-7i-5p inhibitor group. CCK-8 method and clone formation experiment were used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis. Transwell array was used to detect the cell migration and invasion. Western blot was used to detect the protein expressions of Caspase-3, Caspase-9, Bcl-2 and Bax. The double luciferase reporter gene experiment was used to verify the regulatory relationship between DRAIC and let-7i-5p. Independent sample t test was used for comparison between two groups, one-way ANOVA was used for comparison between multiple groups, and Pearson correlation analysis was used for correlation analysis. Results: Compared with adjacent tissues, the expression level of DRAIC in lung adenocarcinoma tissues increased (P<0.05), but the expression level of let-7i-5p decreased (P<0.05). The expression levels of DRAIC and let-7i-5p in lung adenocarcinoma tissues were negatively correlated (r=-0.737, P<0.05). The absorbance value of A549 and H1299 cells in the si-DRAIC group at 48, 72 and 96 hours were lower than those in the si-NC group (P<0.05), the number of clones formed [(91.00±6.08 vs. 136.67±6.51); (50.67±1.53 vs. 76.67±4.51)], the number of migration [(606.67±31.34 vs. 960.00±33.06); (483.33±45.96 vs. 741.67±29.67)], the number of invasion [(185.00±8.19 vs. 447.33±22.05); (365.00±33.87 vs. 688.00±32.97)] were lower than those in the si-NC group (P<0.05). However, the apoptosis rates of cells [(13.43±2.79)% vs. (4.53±0.42)%; (23.77±1.04)% vs. (6.60±1.42)%] were higher than those in the si-NC group (P<0.05). The protein expressions of Caspase-3, Caspase-9 and Bax in si-DRAIC group were higher than those in si-NC group, and the protein expression of Bcl-2 was lower than that in si-NC group (P<0.05). DRAIC is located in the cytoplasm. DRAIC targeted and negatively regulated the expression of let-7i-5p. The absorbance values of A549 and H1299 cells in the let-7i-5p mimics group at 48, 72 and 96 hours were lower than those in the miR-NC group (P<0.05), the number of clones formed [(131.33±14.47 vs. 171.33±6.11); (59.33±4.93 vs. 80.33±7.09)], the number of migration [(137.67±3.06 vs. 579.33±82.03); (425.00±11.14 vs. 669.33±21.13)], the number of invasion [(54.00±4.36 vs. 112.67±11.59); (80.00±4.58 vs. 333.33±16.80)] were lower than those in the miR-NC group (P<0.05). However, the apoptosis rates of cells [(14.57±1.10)% vs. (6.97±1.11)%; (23.97±0.42)% vs. (7.07±1.21)%] were higher than those in the miR-NC group (P<0.05). The protein expressions of Caspase-3, Caspase-9 and Bax in let-7i-5p mimics group were higher than those in miR-NC group, and the protein expression of Bcl-2 was lower than that in miR-NC group (P<0.05). The absorbance values of A549 and H1299 cells in the si-DRAIC+ let-7i-5p inhibitor group at 48, 72 and 96 hours were higher than those in the si-DRAIC+ inhibitor-NC group (P<0.05), the number of clones formed [(82.00±5.29 vs. 59.00±5.57); (77.67±4.93 vs. 41.33±7.57)], the number of migration [(774.33±35.81 vs. 455.67±19.04); (569.67±18.72 vs. 433.67±16.77)], the number of invasion [(670.33±17.21 vs. 451.00±17.52); (263.67±3.06 vs. 182.33±11.93)] were higher than those in the si-DRAIC+ inhibitor-NC group (P<0.05). However, the apoptosis rates of cells [(7.73±0.45)% vs. (19.13±1.50)%; (8.00±0.53)% vs. (28.40±0.53)%] were lower than those in the si-NC group (P<0.05). The protein expressions of Caspase-3, Caspase-9 and Bax in si-DRAIC+ let-7i-5p inhibitor group were higher than those in si-DRAIC+ inhibitor-NC group, and the protein expression of Bcl-2 was lower than that in si-DRAIC+ inhibitor-NC group (P<0.05). Conclusion: DRAIC is highly expressed in lung adenocarcinoma, and DRAIC promotes the proliferation, migration and invasion of lung adenocarcinoma cells and inhibits apoptosis by targeting let-7i-5p.

Association between atmospheric particulate matters and outpatient visits for respiratory disorders in Jiaxing City of Zhejiang Province from 2019 to 2021： a time series analysis / 上海预防医学

ObjectiveTo explore the effect of exposure to atmospheric particulate matters on the outpatient visits of respiratory disorders in Jiaxing City，Zhejiang Province. MethodsDaily air pollutant monitoring data，meteorological data and outpatient visits of respiratory disorders in Jiaxing City from 2019 to 2021 were collected．A generalized additive model was applied to evaluate the effect and laggeel effect of the concentrations of atmospheric particulates for outpatient visits of respiratory disorders after adjusting for secular trend， day-of-the-week effect， holiday effect， and meteorological variables. ResultsThe daily average concentrations of PM2.5， PM10， O3 and NO2 exceeded the standard， and the proportion of days exceeding the standard was 3.4%， 1.3%， 11.0% and 0.8%， respectively. Every 10 μg·m-3 increase in PM2.5 concentration showed the strongest effects on the daily outpatient visits of respiratory disorders， adult and childhood respiratory disorders all on lag07 with ER（95%CI） being 2.29%（1.35%‒3.24%）， 2.31% （1.39%‒3.23%） and 2.65 % （1.36%‒3.96%）， respectively. The maximum ER of outpatient visits for respiratory disorders in children was higher than that in adults. Every 10 μg·m-3 increase in PM10 concentration showed the strongest effects on the daily outpatient visits of respiratory disorders on lag07， adult respiratory disorders on lag06 and childhood respiratory disorders on lag07 with ER（95%CI） being 1.42% （0.87%‒1.96%）， 1.49%（0.99%‒1.99%） and 1.61% （0.87%‒2.36%）， respectively. The results of double-pollutant model showed that the effect of atmospheric particulate reduced after O3 was introduced into the model. ConclusionThere are a short-term effect and a laggeel effect of atmospheric particulate on the outpatient visits of respiratory disorders. It is necessary to strengthen the health protection of the respiratory system of the population， especially the children.

Time trend of pesticide poisoning in Jiaxing from 2008 to 2020 / 环境与职业医学

Background Pesticide poisoning is not only a common acute poisoning, but also an indispensable public health problem. It is important to describe and analyze the epidemic characteristics and trends of pesticide poisoning for its prevention and control. Objective To understand the epidemiological characteristics and trends of pesticide poisoning in Jiaxing from 2008 to 2020, and provide a basis for making effective intervention measures. Methods The relevant information of pesticide poisoning cases in Jiaxing from 2008 to 2020 was collected through the Occupational Disease and Occupational Health Information Monitoring System of the China Disease Prevention and Control Information System, and the demographic information was obtained from the statistical yearbook of Jiaxing. Joinpoint regression models were used to analyze trends in overall, gender, age, season, type of poisoning, and type of pesticide among poisoned individuals. Results A total of 3109 cases of pesticide poisoning were reported in Jiaxing City from 2008 to 2020. The overall pesticide poisoning incidence trended downward from 2008 to 2014, with an annual percent change (APC) of −9.0% (95%CI: −16.6%-−0.7%). The female pesticide poisoning incidence trended downward from 2008 to 2015, with an APC of −8.6% (95%CI: −13.9%-−2.9%). The 18-34 age group showed a decreasing trend of pesticide poisoning incidence from 2008 to 2015, with an APC of −11.0% (95%Cl: −17.4%-4.3%), and an increasing trend from 2015 to 2020, with an APC of 18.5% (95%Cl: 4.7%-34.0%). The >60 age group showed a decreasing trend of pesticide poisoning incidence from 2008 to 2014, with an APC of -12.9% (95%Cl: −20.4%-−4.7%). The second quarter showed an increasing trend of pesticide poisoning incidence from 2010 to 2020, with an APC of 4.4% (95%CI: 0.3%-8.5%); the third quarter showed a decreasing trend, with an APC of −4.9% (95%CI: −8.6%-−1.1%); the fourth quarter showed an increasing trend from 2015 to 2020, with an APC of 17.8% (95%CI: 4.4%-33.0%). Productive poisoning showed a decreasing trend, with an APC of −11.1% (95%CI: −16.2%-−5.7%); self-poisoning showed a decreasing trend from 2008 to 2014, with an APC of -9.5% (95%CI: −17.4%-−0.7%), and an increasing trend from 2014 to 2020, with an APC of 10.2% (95%CI: 0.5%-20.8%). The incidences of poisoning by herbicides, fungicides, and mixed formulations all showed an increasing trend from 2008 through 2020, with an APC of 8.6% (95%CI: 5.8%-11.5%), 9.1% (95%CI: 0.3%-18.7%), and 193.3% (95%CI: 11.6%-671.0%), respectively; the incidence of poisoning by other types of pesticides showed a decreasing trend from 2008 to 2020, with an APC of −14.1% (95%CI: −23.7%-−3.2%). Conclusion The overall reported pesticide poisoning incidents in Jiaxing City present a decline then a rise in 2008 to 2020. Relevant departments should take timely measures to prevent and reduce the occurrence of pesticide poisoning according to the changing characteristics and occurrence trends of local pesticide poisonings.

Research progress of traditional Chinese medicine formulas and active compounds in the treatment of non-alcoholic fatty liver disease by regulating gut microbiota / 药学学报

Non-alcoholic fatty liver disease (NAFLD) is one of the most common chronic liver diseases. However, due to its complex pathogenesis, there are no officially approved drugs for NAFLD treatment currently. Therefore, it is extremely urgent to find safe and effective anti-NAFLD drugs. Nowadays, lipid-lowering drugs are the main option for NAFLD therapy, but the clinical efficacy of chemical drugs is also very limited, as well as the frequent side effects or adverse reactions. Traditional Chinese medicine (TCM) has attracted more and more attention in the treatment of NAFLD due to its unique advantages through multiple targets and pathways with few side effects. In recent years, numerous studies have demonstrated that the imbalance of gut microbiota plays an important role in the occurrence and development of NAFLD. This review systematically summarizes the experimental and clinical evidences of TCM active compounds and TCM prescription involved in the regulation of intestinal flora in the treatment of NAFLD in recent years, so as to provide a reference for further exploring the pathogenesis of NAFLD and exploring TCM treatment methods.

Targeted therapy for malignant peripheral nerve sheath tumor: translational research and clinical application / 中华肿瘤杂志

Malignant peripheral nerve sheath tumor (MPNST) is a rare invasive soft tissue sarcoma that originates from peripheral nerve branches and peripheral nerve sheaths. Early radical surgery is an effective treatment for MPNST. Since it is insensitive to radiotherapy and chemotherapy, the disease manifests a rapid progression, poor prognosis and high mortality. In recent years, the translational researches on the driving factors and therapeutic targets of MPNST have been rapidly developed, including the pathways of NF1-Ras, Raf-MEK-ERK, PI3K-AKT-mTOR, Wnt signaling, and abnormal expressions of apoptotic proteins, the general loss of polycomb repressive complex 2 (PRC2), upregulation of the HDAC family, abnormal expressions of receptor tyrosine kinases, expressions of programmed cell death ligand (PD-L1), aurora kinase and various microRNAs.This review summarizes the current translational researches on potential therapeutic targets of MPNST, and the clinical trials which provide helpful information for MPNST targeted therapy.

Consistency Analysis of Gene Mutation Sites of Bone Marrow Tumor DNA and Circulating Tumor DNA in Patients with Myelodysplastic Syndrome / 中国实验血液学杂志

OBJECTIVE@#To analyze the consistency of gene mutation sites between bone marrow DNA (BM-tDNA) and perepheral plasma circulating tumor DNA (PP-ctDNA) in patients with myelodysplastic syndrome (MDS).@*METHODS@#The simultaneous sampled BM and PP from 19 patients (SBPP) was detected by NGS-127 gene panel, and the consistency of VAF between BM-tDNA and PP-ctDNA was analyzed. The peripheral blood cell tumor DNA (PC-tDNA) of 5 out of 19 patients was detected randomly, the consistency of VAF among PC-tDNA，BM-tDNA and PP-ctDNA was analyzed. The non simultaneous sampled BM and PP from 13 patients (NBPP) was detected, and the difference value of VAF between BM-tDNA and PP-ctDNA in SBPP and NBPP was analyzed.@*RESULTS@#The average concentration of PP-ctDNA in SBPP was 0.59 ng/µl and 0.604 ng/µl in NBPP. The median concentration of PP-ctDNA in SBPP and NBPP was 0.330 ng/µl and 0.338 ng/µl, respectively. The study showed a good consistency of VAF between BM-tDNA and PP-ctDNA in the SBPP (R=0.9693, P<0.05), and the consistency of VAF between BM-tDNA and PP-ctDNA in single base replacement (SNP) sites (R=0.9712) was better than that in insertion deletion (Indel) sites (R=0.6813). The results showed a good consistency of VAF between BM-tDNA and PP-ctDNA both in 12 patients before treatment (R=0.9325, P<0.05) and 5 patients (R=0.9875, P<0.05) after treatment. The results also showed that the VAF of PC-tDNA had a good consistency with the VAF of BM-tDNA (R=0.8783) and PP-ctDNA (R=0.8783) (P<0.05). The difference value of VAF between BM-tDNA and PP-ctDNA in SBPP was significantly lower than that in NBPP (P<0.05).@*CONCLUSION@#PP can replace BM as a biological sample for genes mutation detection in patients with MDS due to its stable concentration, high degree of consistency with bone marrow in clinical significant mutation sites and easy collection.

Relationship between the expression of Cofilin in alveolar and peripheral blood mononuclear cells in COPD patientsand the function of phagocytic aspergillus / 实用医学杂志

Objective To investigate the expression of Cofilin in alveolar and peripheral blood mononuclear cells in COPD patients and the correlation with the function of phagocytic aspergillus. Methods 20 COPD pa-tients were selected from July 2015 to May 2016 in the outpatient department of respiratory medicine of Zhujiang Hospital of Southern Medical University ,then divided into the experimental group 1(group Aand B in 2011 version of GOLD),group 2(test group C and group D in 2011 version of GOLD),and the healthy control group in 10 people. The AM and MDM in the peripheral bloodwere extracted respectively in the 3 groups by bronchoalveolar lavage and,and the ability of AM and MDM in each group were detected. The expression of Cofilin protein was measured by real-time fluorescence quantitative(qRT-PCR) and Western blotting ,and the differenceswere compared among the three groups. Results The colony numbers of MDM/AM in the 3 groups were 17 ± 3,16 ± 2, 42 ± 3(F = 73.446 ,P < 0.001),and the colony numbers of MDM/AM in the two test groupshad significantly different from the healthy group ,while no significant difference was found in the two test groups. The results of fluorescence quantitative reverse transcription polymerase chain reaction and Western blot analysis showed that the expression of Cofilin in the two test groups was significantly higher than that in the healthy group. Conclusion The decreased function of phagocytic aspergillus in the alveolar and peripheral blood mononuclear cells in COPD patients may relates to the increase of cofilin.

Application of IMA and H-FABP in Forensic Diagnosis of Sudden Cardiac Death / 法医学杂志

Acute myocardial ischemia is the most common cause of sudden cardiac death.The diagnosis of early myocardial ischemia is a hot point in forensic medicine,which is also an early and important part for a prevention against myocardial infarction.This paper conducts a comprehensive discussion of the structure,function,clinical value and forensic medicine application prospect of ischemia modified albumin (IMA) and heart-type fatty acid binding protein (H-FABP),aiming to determine whether the two proteins can be used as biochemical detection indicators of early myocardial ischemia for the diagnosis of sudden cardiac death in forensic medicine.

Application of Ischemia Modified Albumin for Acute Ischemic Heart Disease in Forensic Science / 法医学杂志

Objective To explore the application value and forensic significance of ischemia modified albumin (IMA) in pericardial fluid to diagnose sudden cardiac death.Methods IMA level in pericardial fluid was detected in acute ischemic heart disease group (n=36),acute myocardial infarction group (n=6),cardiomyopathy group (n=4) and control group (n=15) by albumin cobalt binding method.The levels of IMA were compared among these groups.The best cut-off IMA value was estimated and the sensitivity and specificity of acute myocardial ischemia group was distinguished from control group by receiver operating characteristics (ROC) curve.Results The IMA level in acute ischemic heart disease group was significantly higher than that of control group (P＜0.05).Compared with acute myocardial infarction group and cardiomyopathy group,the IMA level in acute ischemic heart disease group had no significant difference (P＜0.05).The cut-off value for the identification of acute myocardial ischemia which obtained by ROC analysis was 40.65U/mL.And the sensitivity and specificity for distinguishing acute ischemia cardiac disease was 60.0％ and 80.5％,respectively.Conclusion The IMA value in pericardial fluid can be a reference marker for the diagnosis of acute myocardial ischemia,which also can provide objective basis for the forensic identification of sudden cardiac death.

Construction of JSRV-NM Pseudovirions by High Efficiency Packaging System of the Lentivirus / 天津医药

Objective To overcome the fact that SRV-NM virus can only multiple and amplify through partially pu-rified jaagsiekte retrovirus inoculated intratracheally in sheep but it cannot be augmented using in vitro cell culture, we con-structed JSRV-NM pseudovirions based on high efficiency packing system of lentivirus. Methods Lentivirus of three high efficiency packing plasmids system pMD.G, pCMV-HIV 8.2 and pHIV-eGFP was developed, and JSRV-NM-env coated plasmid pCMVJSRV-NM was used to substitute VSV-G virus coated plasmid pMD.G then co-transfected into 293T cells to replicate, package and produce restructured JSRV-NM pseudovirions. Gene expression of pseudovirion was determined through WPRE using real time PCR; Virus infectivity was detected through inoculating JSRV-NM pseudovirions into 24 pore plates. Results We construct JSRV-NM pseudovirions successfully based on the lentivirus system. JSRV-NM pseudo-virions can also be concentrated to higher titer (108 TU/mL detected by real time PCR by ultracentrifugation without signifi-cant loss of activity. JSRV-NM and VSV-G pseudovirions infected on Hela cells (both MOI= 3) respectively and no obvi-ous difference were shown on their infection efficiency detected by real time PCR. Conclusion Based on lentivirus system, JSRV-NM pseudovirions can be multipled and amplified in 293T cell culture in vitro. JSRV-NM pseudovirions is stable without loss its infection activity and the requirements of biological laboratory safety II was also met. JSRV-NM pseudoviri-ons will provide a useful tool for further study of JSRV-NM-env infection across species or its induction of lung adenocarci-noma.

Construction and identification of a mouse spermatocyte-derived cell line with a stable expression of PIAS-NY / 中华男科学杂志

<p><b>OBJECTIVE</b>To construct a lentiviral expression vector of the PIAS-NY gene, and establish a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY.</p><p><b>METHODS</b>PIAS-NY was synthesized, amplified by PCR and cloned into the lentiviral vector expression plasmid pGC-FU. After digestion and sequencing, pGC-FU-PIAS-NY, pHelper 1.0 and pHelper 2.0 were co-transfected into 293T cells. Then the lentiviral particles were used to transfect the mouse spermatocyte-derived cells. The expression of the PIAS-NY protein was detected by Western blot.</p><p><b>RESULTS</b>We successfully constructed the lentiviral expression vector pGC-FU-PIAS-NY and established a mouse spermatocyte-derived cell line with a stable overexpression of PIAS-NY.</p><p><b>CONCLUSION</b>The construction of the lentiviral expression vector pGC-FU-PIAS-NY and the obtainment of stably transfected mouse spermatocyte-derived cells have paved the way for further studies on the roles of the PIAS-NY gene in spermatogenesis.</p>

Expression and significance of mRNA and protein of eIF4E, p-eIF4E and MCl-1 in pathological scar / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the expression of eIF4E, p-eIF4E (Ser 209) and Mcl-1 gene in the pathological scars and to investigate its role and its probable mechanism in the pathogenesis of abnormal scar.</p><p><b>METHODS</b>Quantitative real-time PCR and Western Blot was performed to detect the expression and distribution of mRNA and protein of eIF4E and Mcl-1 in hypertrophic scar (10 cases), keloid (10 cases), normal scar (10 cases), and normal skin (10 cases). Western Blot was performed to detect the expression and distribution of protein of p-eIF4E in hypertrophic scar (10 cases), keloid (10 cases), normal scar (10 cases), and normal skin (10 cases).</p><p><b>RESULTS</b>The expression of eIF4E mRNA and protein were 1.38 +/- 0.45, 1.23 +/- 0.23 in the normal skin (10 cases); 5.400 +/- 0.450, 5.460 +/- 0.460 in normal scar (10 cases); 0.597 +/- 0.060, 0.590 +/- 0.040 in hypertrophic scar (10 cases) and 0.694 +/- 0.066, 0.697 +/- 0.022 in keloid (10 cases). The expression of p-eIF4E protein in the normal skin (10 cases), normal scar (10 cases), hypertrophic scar (10 cases), and keloid (10 cases) were 0.202 +/- 0.037, 0.216 +/- 0.019, 0.426 +/- 0.026, 0.433 +/- 0.027. The expression of Mcl-1 mRNA and protein were 1.510 +/- 0.660, 1.400 +/- 0.530 in the normal skin (10 cases); 6.65 +/- 0.85, 7.23 +/- 1.53 in normal scar (10 cases); 0.589 +/- 0.059, 0.660 +/- 0.063 in hypertrophic scar (10 cases) and 0.870 +/- 0.118, 0.914 +/- 0.064 in the keloid (10 cases). The positive rate of mRNA and protein of eIF4E and Mcl-1 was not statistically different between the hypertrophic scar and keloid (P > 0.05), while they were all remarkably significant between normal scar and abnormal scar (P < 0.05). The phosphorylation of eIF4E in pathological scar was higher than that in control group. In pathological scar, mRNA and protein of eIF4E and Mcl-1 showed a strong positive correlation.</p><p><b>CONCLUSIONS</b>The result indicates that the expression of eIF4E, p-eIF4E and Mcl-1 is increased in pathological scar. eIF4E plays an important role in pathological scar. Its activity is regulated by its phosphorylation. Therefore, eIF4E, p-eIF4E and Mcl-1 overexpression may play an important role in the proliferation of fibroblasts and in the pathogenesis of pathological scar.</p>

Expression and significance of P57(kip2) and Maspin in pathological scar / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the expression of P57(kip2) and Maspin in the pathological scar and their possible role in the pathogenesis of abnormal scars.</p><p><b>METHODS</b>Immunohistochemistry integrated image analysis and reverse transcription polymerase chain reaction (RT-RCR) were performed to detect the expression of P57(kip2) and Maspin in hypertrophic scar, keloid, mature scar and normal skin. Statistics was used to analyze the datas.</p><p><b>RESULTS</b>The expression of P57(kip2) protein was fixed to fibroblast intranuclear in abnormal scar, and the expression of P57(kip2) protein and P57(kip2) mRNA decreased (P < 0.05). The expression of Maspin protein was fixed to fibroblast cytoplasm and intranuclear in abnormal scar, and the expression of Maspin protein and Maspin mRNA decrease, compared with that in normal group (P < 0.05). There was positive correlation between P57(kip2) protein and Maspin protein expression (P < 0.01).</p><p><b>CONCLUSIONS</b>The decreased expression of P57(kip2) and Maspin in abnormal scar shows that they are cicatrix-related genes. There is a positive relationship between the two genes. It may be one of the mechanisms of pathogenesis of abnormal scar. It makes effect through fibroblasts.</p>

Establishment of a fluorescent real-time quantitative RT-PCR assay for detection of genotype 4 hepatitis E virus in swine stools / 病毒学报

The primers and probes for the Real-time RT-PCR were designed based on the multiple sequence (swine and humans HEV strains) alignments of the ORF3 region of genotype 4 HEV. A rapid, sensitive and stable TaqMan Real-time RT-PCR assay was established, and its specificity and sensitivity were assessed, and comparison of the Real-time RT-PCR with conventional and nested RT-PCR was performed. The results found that the crossing points showed linearly proportional to the logarithm of the input copy number. The correlation coefficient (R2) and the slope value of the standard curves with plasmid DNA were 0.994 and -3.312, respectively. The efficiency (E) of the PCR was 100%. Coefficients of variation values of the different diluted plasmid DNA were low in the same or different repeated experimental group. In addition, the assay was able to correctly detect genotype 4 HEV RNA from swine fecal samples. The sensitivity of established assay was 100-fold higher than that of conventional RT-PCR and 10-fold higher than nested RT-PCR.

Correction of severe depression deformities at lower eyelid / 中华整形外科杂志

<p><b>OBJECTIVE</b>To evaluate the application of temporal fascia flaps in the correction of severe depression deformities at lower eyelids.</p><p><b>METHODS</b>Severe depression deformities at lower eyelid were corrected with temporal fascia flaps pedicled with superficial temporal artery in 9 cases.</p><p><b>RESULTS</b>All flaps survived with good appearance. All patients were followed up for 6-12 months with good long-term results. The donor sites had no obvious scalp scar.</p><p><b>CONCLUSIONS</b>Temporal fascia flap is an optimal choice for correction of the severe depression deformities at lower eyelid. It is easily performed with good result and less donor site morbidity.</p>

Susceptibility of Candida albicans to Fluconazole by Rapid Flow Cytometry / 中华医院感染学杂志

0.05) and the two methods had good correlation(r=0.822).CONCLUSIONS The method of FCST established by as in this study is simple,repeatable,with high accuracy and easy to determine MIC and has good application prospects in clinical antifungal susceptibility testing.

Island skin flap supplied by superficial temporal vessel for the whole ear lobe defect / 中华整形外科杂志

<p><b>OBJECTIVE</b>To introduce a new method for reconstruction of the whole ear lobe defect.</p><p><b>METHODS</b>The free island skin flap supplied by superficial temporal vessel which was designed at the area anterior and superior to crus helicis. The flap was transferred through subcutaneous tunnel and self-folded to reconstruct the whole ear lobe defect.</p><p><b>RESULTS</b>Since 1999, 6 cases were treated with no complication. The ear lobe shape and skin colour were very natural.</p><p><b>CONCLUSIONS</b>The island skin flap supplied by superficial temporal vessel is very suitable for the whole ear lobe defect with good cosmetic results.</p>

Expression and significance of Skp2 and p27kip1 protein in pathological scar / 中华整形外科杂志

<p><b>OBJECTIVE</b>To study the expression of Skp2 gene (s-phase kinase associated protein 2) in the pathological scars and its relationship with p27kip1 level and to investigate its role and its probable mechanism in the pathogenesis of abnormal scars.</p><p><b>METHODS</b>Immunohistochemical technique was performed to detect the expression and distribution of Skp2 and p27kip1 protein in hypertrophic scar (42 cases), keloid (18 cases), normal scar (40 cases) and normal skin (50 cases), statistics was used to analyze the data.</p><p><b>RESULTS</b>The positive rate of Skp2 and p27kip1 protein expression was not statistically different between the hypertrophic scar and keloid (P > 0.05), while they were all remarkably significant in comparison between normal scar and abnormal scar (P < 0.05). In pathological scar the protein of Skp2 and p27kip1 showed a strong inverse correlation (P < 0.01).</p><p><b>CONCLUSION</b>The result indicated that the expression of Skp2 was increased and it may lead to decrease p27kip1 level in the hypertrophic scar and keloid, Skp2 overexpression might play an important role in the proliferation of fibroblasts and in the pathogenesis of pathological scar by adjusting the regulation of cyclins such as p27kip1.</p>

Cloning and Overexpression of Phytase Gene appA form Escherichia coli / 微生物学通报

A strain Escherichia coli with high production of phytase was screened from pig excreta. Phytase gene appA, with 1,299 bp coding region in full length, was cloned from its genome by polymerase chain reaction (PCR) . The gene appA was then cloned into the prokaryotic expression vector pET-28a ( + ) . In the host BL21, the phytase appA was overexpressed by shaker-cultivation (up to 692 U/mL) . The enzymatic analysis of the prokaryotic derived appA phytase revealed that its optimal pH and temperature was 4.5 and 60℃, respectively.