Environmental Microbiology is an important basic course of Environmental Engineering.Its characteristic is content broad,quick development and strong practicality,thus this curriculum's teaching has certain degree of difficulty.Some suggestions and concrete measures about teaching reform,which included curriculum's course content,teaching method,experiment teaching and assessment methods were proposed in this paper.

Aim To observe the changes of diaphragm contractility and cytoskeletal proteins titin,nebulin and sarcoplasmic reticulum Ca~(2+)-ATPase gene expressions in adriamycin-induced cytotoxicity in rats.Methods The animal models of diaphragm damage were duplicated by injecting adriamycin into abdominal cavity one time.Forty male sprague-dawley rats were randomly divided into four groups(n=10):Three groups received adriamycin in low,middle and high dosage(10,20 and 40 mg·kg~(-1))respectively.Meanwhile,the normal saline was given to rats in control groups.Three days later,these rats were killed,and the diaphragm was removed by thoracotomy.The diaphragm contractility was assessed in isolated diaphragm strips perfusion by these paramemters including peak twitch tension(Pt),maximum tetanic tension(Po),time to peak contraction(CT),half relaxaion time(1/2RT),maximal rates of contraction(+dt/dt_(max))and maximal rates of relaxation(-dt/dt_(max)).The expressions of titin,nebulin and sarcoplasmic reticulum Ca~(2+)-ATPase(SERCA)at mRNA level were detected by RT-PCR analysis.Results In contrast to those in control group,Po,Pt,±dt/dt_(max) in the adriamycin group were lower(P<0.01);CT,1/2RT in the adriamycin group increased significantly(P<0.01).The levels of titin,nebulin and SERCA gene expressions in middle-dose group were lower than those in control group(P<0.01).Conclusions The mRNA levels of titin,nebulin and SERCA of diaphragm are down-regulated in adriamycin-induced cytotoxicity in rats.It may be associated with the decline of diaphragm contractility.

Objective To explore the effects and mechanism of cigarette smoke extract (CSE) on the proliferation of air?way smooth muscle cells (ASMCs) and the expression of CCAAT/enhancer-binding protein (CEBPα) and calreticulin. Meth?ods (1) The ASMCs were stimulated with different concentrations of CSE for twenty-four hours. According to the concentra?tions of CSE,the cells were divided into control group, 2.5%CSE group, 5%CSE group and 10%CSE group. The prolifera?tion of ASMCs was measured by MTT colrimetric method. The CEBPαmRNA was analyzed by RT-PCR. Western bloting as?say was performed to detect the levels of CRT and CEBPαprotein. (2) In 10%CSE group, transfection of the siRNA respec?tively for negative control or calreticulin was performed in accordance with instructions. The cell proliferation and the expres?sion of calreticulin and CEBPαwere compared in negative control siRNA group and calreticulin siRNA group. Results (1) With the increasing of the concentrations of CSE, the protein expression of CEBPαdecreased gradually (P<0.05), while the proliferation of ASMCs and the protein expression of calreticulin increased (P<0.05), but the expression of CEBPαmRNA in ASMCs showed no significant difference in groups with different concentrations of CSE (P>0.05). (2) Under the 10%CSE, the expression of CEBPαwas significantly higher in CRT siRNA group than that in negative control group (P<0.05),but the cell proliferation and CRT were significantly lower in the calreticulin siRNA group than those in negative control siRNA group (P<0.05). Conclusion The CSE exposure contributes to the expression of calreticulin protein,and then inhibits the translation of CEBPαmRNA,thus promotes the proliferation of ASMCs.

Objective To evaluate the value of transvaginal ultrasound in diagnosing intrauterine adhesions.Methods Transvaginal ultrasound was performed in 136 patients with suspicious intrauterine adhesions and compared with hysteroscopy correspondingly .The ultrasonographic features of intrauterine adhesions on transvaginal ultrasound were summarized .Results One hundred and twenty one cases (89.0%, 121/136 ) of intrauterine adhesions were verified by hysteroscopy .The hysteroscopic findings included:(1) Forty seven cases(38.9%,47/121) were minimal intrauterine adhesions , 46 cases(38.0%, 46/121) were moderate intrauterine adhesions , and 28 cases (23.1%,28/121) were severe intrauterine adhesions.(2) Sixty one cases(50.4%,61/121) were central intrauterine adhesions , 24 cases(19.8%, 24/121) were marginal intrauterine adhesions , and 36 cases (29.8%, 36/121) were mixed type of intrauterine adhesions.The transvaginal ultrasound findings included:(1)Nineteen cases(40.4%,19/47) were minimal intrauterine adhesions ,33 cases(71.7%,33/46)were moderate intrauterine adhesions ,and 23 cases(82.1%,23/28) were severe intrauterine adhesions .(2) Thirty nine cases (63.9%,39/61) were central intrauterine adhesions ,9 cases(37.5%,9/24) were marginal intrauterine adhesions ,and 27 cases (75.0%, 27/36 ) were mixed type of intrauterine adhesions .By transvaginal ultrasound, seventy-five (62.0%,75/121) cases of intrauterine adhesions were correctly diagnosed , whereas 46 cases (38.0%, 46/121) were missed.And 3 cases ( 3.8%, 3/78 ) were misdiagnosed as intrauterine adhesions on transvaginal ultrasound,including one endometrial polyp ,one thin endometrium and one septate uterus .The sensitivity, specificity and accuracy of transvaginal ultrasound in diagnosing intrauterine adhesions were 62.0%(75/121), 80.0%(12/15) and 64.0%(87/136) respectively.There were significant statistical differences in diagnosing different degrees of intrauterine adhesions ( χ2 =15.956,P=0.000) and different parts of intrauterine adhesions( χ2 =8.792,P=0.012) by transvaginal ultrasound.Conclusions Transvaginal ultrasound is an effective, easy to perform and noninvasive technique in screening and diagnosing intrauterine adhesions.Transvaginal ultrasound is an effective way in diagnosing intrauterine adhesions showing a noninvasive and simpler way than hysteroscopy .Transvaginal ultrasound is of great value in screening and diagnosing intrauterine adhesions .

To study the effects of Pingchuan Recipe, a compound traditional Chinese herbal medicine for treating bronchial asthma, on macrophage inflammatory protein-1alpha (MIP-1alpha) and immunoglobulin E (IgE) contents and CD86 expression in a mouse model of bronchial asthma, and to investigate the mechanism of Pingchuan Recipe in regulating airway remodeling in mice with bronchial asthma.

Aim To investigate the role of mitochondrial aldehyde dehydrogenase 2 ( ALDH2) in the cardio-protection of ischemic postconditioning in isolated rat hearts. Methods Hearts isolated from male Sprague-Dawley rats were perfused on a langendorff apparatus and subjected to 30 min of regional ischemia( occlusion of left anterior descending artery) followed by 120 min reperfusion. Ischemic postconditioning was achieved by 6 cycles of 10 s reperfusion/10 s global ischemia starting at the beginning of reperfusion. The ventricular hemodynamic parameters and lactate dehydrogenase ( LDH) release during reperfusion were measured. The infarct size was measured by TTC staining method. The expressions of ALDH2,Bcl-2 and Bax at mRNA level of left anterior myocardium were detected by RT-PCR analysis. Results In contrast to ischemia and reperfusion,ischemic postconditioning improved the recovery of left ventricular developed pressure,rate pressure product during reperfusion,and reduced LDH release and infarct size. The expressions of ALDH2 mRNA level and the ratio of Bcl-2 /Bax were increased. Adminis-tration of ALDH2 antagonist cyanamide at the beginning of reperfusion attentuated the role of ischemic postconditioning. Conclusion Ischemic postconditioning plays a role in the cardioprotection partially through increasing mitochondrial ALDH2 mRNA expression.

Objective To investigate the relationship between EVI1 gene expression and clinical features and prognosis of children with acute myeloid leukemia (AML). Methods EVI1 gene was detected in AML children, correlation of clinical and lab features, prognosis of AML children with EVI1 gene were analyzed. Results EVI1 expression is positive in 38 of 145 children with AML. There were no significant differences in age, gender, hemoglobin concentration, leukocytes and platelet count, subtype of morphology, ratio of chromosomal anomaly and complex karyotypes between EVI1 positive and EVI1 negative group (P>0.05); coexist genes were detected in 9 cases (23.68%) of EVI1 positive group. Rate of complete remission (CR) was 91.67% in 24 cases of EVI1 positive patients received chemotherapy. Relapse rate was 64.29% and 14.29% in EVI1 positive patients who received chemotherapy and allo-hematopoietic stem cell transplantation (allo-HSCT), retrospectively and significant differences were found (P<0.05). There was no significant difference in CR but significant difference was found in event free survival (P<0.05) for EVI1 positive and EVI1 negative patients who received chemotherapy. EVI1 gene kept negative when bone marrow relapse occurred in two patients with EVI1 positive at diagnosis. Conclusion EVI1 gene may play adverse role in pediatric AML; prognosis of EVI1 positive AML patients can be improved by allo-HSCT; follow-up of EVI1 transcript levels is insufficient to monitoring of minimal residual disease.

Objective To establish the cell model of ethanol-induced liver injury and explore the protective effects of tea poly-phenols (TP)on ethanol-induced liver injury .Methods Cell morphology were observed by microscope ,and then alanine aminotrans-ferase (ALT) ,nmda transaminase (AST) ,gamma GGTP ,GGT and ROS changes were detected .Results Alcohol maked L02 hepa-tocyte fatty degeneration .Compared with ethanol group ,steatosis in TP + ethanol group was lighter ,its ALT ,AST ,GGT content and intracellular ROS reduced .Conclusion TP can decrease cell fatty change degree in vitro experiments ,improue the enzymology indexes ,reduce the generation of reactive oxygen species to avoid liver damage .

Objective To evaluate the treatment of current status and prognosis in childhood APL with APL2008 ,which was administrated since 2008 in our center .Methods A total of 43 children with newly diagnosed APL between 2008 to 2014 were studied retrospectively .Treatment options and current status were summarized from 28 patients who received APL2008 therapy . Results Studied 43 patients were at median age of 8 years and 4 months ,with 28 boys and 15 girls .The main clinical manifestations were infection ,anemia ,bleeding ,fever ,hepatomegaly ,splenomegaly and lymphadenopathy .The proportions of low ,intermediate and high risk groups were 27 .9% ,48 .8% and 23 .3% ,respectively .Eleven cases could be diagnosed as DIC .Bone marrow morphology showed abnormal elevation of promyelocyte .37 patients had distinctive immunophenotype such as frequent expression of CD33 , CD117 and MPO .PML/RARαfusion gene positive rate was 100% in 43 children and cytogenetic analysis were positive in 37 cases , of which specific genetic lesion in APL cells with t (15 ;17)(q22 ;q12) was found in 28 cases ,and karyotypes was found in 9 cases as infrequent chromosomal abnormalities .In 43 patients ,4 cases were early dead from intracranial hemorrhage at early stage ,and 11 cases were given up early .There were only 2 cases dead ,2 cases relapsed and 1 case lost among 28 APL children ,which enabled ef‐ficacy analysis possible .96 .4% of these 28 cases achieved HCR .The 2 year Kaplan Meier estimates of OS and EFS were 85 .9% ± 7 .6% and 80 .4% ± 8 .8% .But OS and EFS would be 94 .7% ± 5 .1% and 88 .9% ± 7 .4% if 3 patients who had non standard treat‐ment were excluded .Conclusion Childhood APL were characterized by anemia ,bleeding ,fever and infiltration .APL′s coincidence rate between PML/RARa fusion gene and morphology ,immunology and cytogenetics were 95 .3% ,90 .2% and 86 .5% ,respective‐ly .APL2008 significantly improved the prognosis of APL .

Objective To understand the ABO blood type distribution of Buyi and Shui ethnic populations in Libo county of Guizhou province .Methods Totally 726 Buyi and 163 Shui individuals who were unrelated within three generations were detected ABO blood stype with glass-clotted method .Results The ABO blood type distributions in the Buyi and Shui ethnic populations both were in line with the Hardy-Weinberg equilibrium .The Buyi ethnic population ABO phenotypic distribution and gene frequen-cies were O>B>A>AB and r>q>p (r=0 .676 9 ,q=0 .176 7 ,P=0 .146 4) and the national index was 0 .842;while the Shui eth-nic population ABO phenotypic distribution and gene frequencies were O>A>B>AB and r>p>q(r=0 .522 6 ,q=0 .104 0 ,P=0 .164 7) and the national index was 1 .529 .Conclusion The Buyi and Shui ethnic populations′ABO blood type distribution in Libo county of Guizhou province is basically consistent with that of the same ethnic populations in other regions of Guizhou Province of the same ethnic groups .The genetic distance analysis prompts that the regional and ethnic differences exist in ABO blood type dis-tribution .