Microbiology experiment existing independently from microbiology theoretical curriculum is an indispensable compulsory course in contemporary life science. This article presents the principle applied by the National Excellent Microbiology Course teaching group in Nankai University, which is to strengthen the undergraduates’ basic skills of conducting microbiology experiments. With an aim to enhance the core competitiveness of the undergraduates, we have established the three-level experimental contents. A new multilevel teaching pattern focusing on basic skill training as the cornerstone has been applied to enhance the overall competences of the students and to stimulate their innovation abilities. Students’ experimental accomplishment will also be taken into consideration when their experiment results are evaluated, which helps to standardizing their research ethics.

MP-Rep fusion gene which size is about 1000bp was constructed by recombinant PCR technique.In order to construct fusion gene,Tobacco mosaic virus(TMV) partial movement protein gene(MP) and Cucumber mosaic virus(CMV) partial replicase gene(rep) were ligated.Two copies of MP-Rep fusion gene were ligated with soybean intron in inverted repeat manner,the recombinant fragments were then inserted into binary vector pBIN438 under the control of 35S promoter.Recombinant clone pBIN438-MP-Rep(i/r) which contained two different virus derived genes was constructed.Recombinant clone pBIN438-MP-Rep(i/r) in accord with expected design was certified by restriction endonuclease enzymes digestion and PCR analysis.This approach provides a basis for Broad-spectrum plant virus resistance mediated by RNA Silencing.

OBJECTIVETo investigate the effect of intensive insulin therapy on resting energy expenditure in postoperative patients underwent radical distal gastrectomy.

METHODSSixty-four patients with gastric neoplasms in the middle or lower part of stomach from January to October 2010 were enrolled and underwent radical distal gastrectomy, then were randomized to intensive insulin therapy (IIT) group to keep glucose levels from 4.4 to 6.1 mmol/L or conventional insulin therapy (CIT) group to keep levels from 4.4 to 10.0 mmol/L. Resting energy expenditure (REE), respiratory quotient (RQ), resting energy expenditure per kilogram (REE/kg) and lipid oxidation ratio (LOR) were monitored by indirect energy metabolic system on preoperative and postoperative 1(st), 3(rd) and 7(th) day. Fasting blood glucose and insulin concentration were measured for HOMA-IR assessment.

RESULTSCompared with preoperative baseline, postoperative REE, REE/kg, LOR, Ln-HOMA-IR increased dramatically (P < 0.05, respectively). RQ decreased markedly (P < 0.05). Compared with group CIT, IIT reduced the REE/kg level [(27.2 ± 1.3) kcal/kg vs. (30.0 ± 1.5) kcal/kg, P = 0.008; (24.7 ± 1.4) vs. (25.7 ± 1.6) kcal/kg, P = 0.013]; and decreased the Ln-HOMA-IR score (P = 0.019 and 0.028) on postoperative 1(st) and 3(rd) day; IIT could decrease obviously the level of C-reaction protein level on postoperative 1(st) and 3(rd) day (P = 0.017, 0.006). The total protein and albumin concentration in IIT group were more than its levels in group CIT (P = 0.023, 0.009).

CONCLUSIONThere are some benefits of IIT in reducing mean energy expenditure and the consumption of proteins, decreasing postoperative insulin resistance level in this small population underwent radical distal gastrectomy.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Energy Metabolism ; Female ; Gastrectomy ; Humans ; Insulin ; administration & dosage ; therapeutic use ; Insulin Resistance ; Male ; Middle Aged ; Postoperative Period ; Stomach Neoplasms ; metabolism ; surgery ; Young Adult

This study is to investigate the effect of the major chemical composition in rhizome of Pterocypsela elata, lactuside B, on expression of bcl-2, bax mRNA and their protein in rats' cerebral cortex after cerebral ischemia-reperfusion injury. First, middle cerebral artery ischemia-reperfusion injury model was established, and each group was treated with the corresponding medicines. Animals were separately sacrificed at 24 h and 72 h. The brain infarct volumes were detected by TTC dye, bcl-2 and bax mRNA expression was checked by RT-PCR, and the proteins of bcl-2 and bax were explored by two-step immunohistochemistry in cerebral cortex of rats. Lactuside B can reduce brain infarct volume of cerebral cortex of rats, increase the expression of bcl-2 mRNA and decrease that of bax mRNA. Moreover, the ratio of bcl-2 to bax mRNA is higher in 12.5 and 25 mg kg(-1) dose group, respectively, which is significantly different from that of model group (P < 0.05 or P < 0.01). Generally, either 12.5 or 25 mg kg(-1) dose group is better than positive control medicine nimodipine (P < 0.05 or P < 0.01). In addition, the expression of bcl-2 and bax protein is consistent with their gene expression. Infarct volume and the ratio of bcl-2 to bax mRNA expression are significantly different (P < 0.05 or P < 0.01) between 72 h and 24 h group. The results demonstrated that lactuside B could play a good role in resisting cerebral ischemia by upregulating the expression of bcl-2 mRNA and protein and downregulating that of bax mRNA and protein.
Animals ; Apoptosis ; drug effects ; Asteraceae ; chemistry ; Brain Ischemia ; metabolism ; pathology ; Cerebral Cortex ; metabolism ; pathology ; Dose-Response Relationship, Drug ; Glucosides ; administration & dosage ; isolation & purification ; pharmacology ; Male ; Neurons ; drug effects ; pathology ; Plants, Medicinal ; chemistry ; Proto-Oncogene Proteins c-bcl-2 ; genetics ; metabolism ; RNA, Messenger ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; metabolism ; pathology ; Rhizome ; chemistry ; Vasodilator Agents ; administration & dosage ; isolation & purification ; pharmacology ; bcl-2-Associated X Protein ; genetics ; metabolism

OBJECTIVETo investigate the influence of cigarette smoking on human sperm DNA integrity.

METHODSTotally, 784 cases of male infertility were selected from our case database and grouped according to whether they were smokers or nonsmokers, how much they smoked (< or = 10, 11-19 and > or = 20 cigarettes/d) and how long they smoked (< or = 5, 6-9 and > or = 10 yr). Sperm DNA integrity was measured using sperm chromatin structure assay (SCSA) and flow cytometry. DNA fragmentation and immature spermatozoa were expressed by the DNA fragmentation index (DFI) and high DNA stainability (HDS) respectively. Conventional sperm parameters and sperm DNA integrity were compared among different groups.

RESULTSThe total semen volume and percentage of grade a + b sperm were lower and the sperm morphological abnormality was higher in the > or = 20 cigarettes/d and > or = 10 yr groups than in the others (P < 0.05). DFI and HDS were significantly higher in the smokers than in the nonsmokers (P < 0.05). HDS was negatively correlated with the percentage of grade a + b sperm (r = -0.18, P < 0.05) and both DFI and HDS were positively correlated with the rate of sperm malformation (r = 0.31 and r = 0.39, P < 0.05).

CONCLUSIONSmoking more than 20 cigarettes a day or longer than 10 years has deleterious effects on the semen volume, percentage of grade a + b sperm and sperm morphology of the smokers. Cigarette smoking decreases sperm DNA integrity and nuclear maturation.

Adult ; DNA Damage ; drug effects ; DNA Fragmentation ; Humans ; Infertility, Male ; genetics ; Male ; Middle Aged ; Smoking ; adverse effects ; Sperm Count ; Sperm Motility ; Spermatozoa ; drug effects ; Young Adult

To investigate the relationship between the plasma biomarker proteins and the states of Zang-Fu organs in patients with phlegm or blood stagnation syndromes due to hyperlipidemia and atherosclerosis.

Objective To evaluate the findings of computed tomography (CT) as objective markers of bone remodeling in rabbit models with chronic rhinosinusitis.Methods Forty-eight rabbit models were 1established by vaccination of staphylococcus aureus.The rabbits were divided into 3 groups according to the time of infection:group A,B and C (4,8,12 weeks after infection),16 rabbits in each group.Each group was subdivided into the medication administration team and the control team,8 rabbits in each team.All the rabbits were examined by CT before vaccination to rule out the disease of nasal cavity and sinuses,and the CT images were used as the negative control.No interference was given to the control teams which were only examined again by CT when reached the end week.Dexamethasone sodium phosphate were administered to the medication administration teams 2 weeks before the end of experiment,and were examined by CT at the end.The images of both horizontal position and coronal position by reconstruction were obtained.The Hounsfield unit (Hu) of the bone which was the thickest position in each image were measured.The data was analyzed by SPSS 16.0 software.The Hu was analyzed statistically to compare the situation of the bone remodeling in different periods and administration in the rabbit models with CRS.Results Average Hu ((x) ±s) of normal rabbits was 810.0 ± 99.7,average Hu at the end time:control team in group A was 964.0 ± 84.6,medication administration team in group A was 833.0 ± 92.5 ; control team in group B was 987.0 ± 91.5,medication administration team in group B was 886.0 ± 91.6; control team in group C was 1086.0 ± 74.0,medication administration team in group C was 899.8 ± 76.5.The Hu in all groups were higher than normal (t value were 2.747,4.5 1 3 and 7.350 respectively,all P ＜ 0.05 ).No statistical difference was found between control teams of group A and B ( t =0.423,P =0.667 ).The Hu in control team of group C was higher than group B ( t =3.905,P =0.001 ) ; There was no statistical difference between medication administration teams of group A and B ( t =0.892,P =0.384 ),and group B and C ( t =0.886,P =0.385 ).The Hu of all medication administration teams in 3 groups were lower than all the control teams ( t value were 2.717,3.687,8.379 respectively,all P ＜ 0.05 ).Conclusions Bone remodeling was found in rabbit models with rhinosinusitis,and the phenomenon was more obvious if the period was lengthened.The Hu could reflect the degree of bone remodelling.Glucocorticoids could depress the bone remodeling in the rabbit models with rhinosinusitis.

OBJECTIVEThe growth and metastasis of solid tumors are dependent on angiogenesis. Endostatin, the C-terminal proteolytic fragment of collagen XVIII, is a potent endogenous angiogenesis inhibitor. The authors designed a topical antiangiogenic gene therapy with recombinant human endostatin adenovirus (Ad-hEndo) and assessed its effects on the inhibition of angiogenesis in vitro, and tumor growth and metastasis in vivo.

METHODSMalignant cells (A549) were infected with Ad-hEndo. The expression of recombinant protein and the inhibition of cultured human umbilical vein endothelial were investigated. Immunodeficient A549 nude mice were treated with intratumoral injection of Ad-hEndo, the empty vector Ad-control or saline (NS). The dose-response, side effects, and serum concentration of endostatin were observed.

RESULTSRecombinant endostatin protein was detected in the infected tumor cells with different MOI Ad-hEndo and its inhibitory effect on endothelial cells growth was shown. In animal study, the volume of tumor and the number of pulmonary metastatic lesions in the Ad-hEndo treatment group were significantly smaller than those in the control groups (P<0.05).

CONCLUSIONThe present findings provide evidence of the anti-tumor effects of the endostatin and may be important for the further use of it in topical antiangiogenic gene therapy of cancer.

Adenoviridae ; genetics ; Animals ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Cells, Cultured ; Disease Models, Animal ; Endostatins ; biosynthesis ; genetics ; therapeutic use ; Endothelial Cells ; Genetic Therapy ; Genetic Vectors ; Humans ; Kidney ; cytology ; metabolism ; Mice ; Mice, Nude ; Neoplasm Metastasis ; Neoplasm Transplantation ; Neoplasms, Experimental ; prevention & control ; Random Allocation ; Recombinant Proteins ; biosynthesis ; genetics ; therapeutic use ; Transfection ; Umbilical Veins ; cytology ; Xenograft Model Antitumor Assays

OBJECTIVETo identify the etiology of an aseptic encephalitis outbreak (ten cases) in a hospital of Xiamen city from 11 to 17 May, 2011.

METHODSA total of ten patients' throat swabs, anal swabs and cerebrospinal fluid were collected and detected by RT-PCR for pan-enterovirus. The samples containing detectable pan-enterovirus were tested by PCR with genotype-specific general primers located in VP1 region of enterovirus genotype A, B and C (HEV-A, B and C). The PCR products of VP1 segment were purified and sequenced, and phylogenetic analysis was performed. Meanwhile, the pathogens in those samples were isolated in Vero cell culture. Homologous analysis of VP1 sequences were carried out for the cultured virus samples and the original clinical samples to identify the outbreak etiology.

RESULTSAmong the ten cases, seven cases were positive for pan-enterovirus nucleic acid. When tested by genotype-specific PCR, the throat and anal swab samples from those 7 patients were positive with HEV-B VP1 primers. Meanwhile, the HEV-B VP1 segments were sequenced and phylogenetic analyzed, which indicated the seven cases were all infected by enterovirus Echo 30. The sequences from those samples had homology of 95.3% - 97.1% with the epidemic strains in Zhejiang, 2004. Out of the seven cases, the sequences of XM2, XM3, XM4, XM8 throat swab samples and XM3, XM6 throat samples showed 99.4% - 100.0% homology which were different from the sequence of XM1, and the homology was 92.8% - 93.4%. Furthermore, the viruses were isolated using Vero cells from XM1, XM2, XM3, XM4 and XM8 throat swab samples, and the VP1 sequence showed more than 99.9% homology with the original specimens.

CONCLUSIONThe local outbreak of aseptic encephalitis was caused by Echo 30 of enterovirus genotype B, and the epidemic strains may have different genetic background.

Child, Preschool ; China ; epidemiology ; Cross Infection ; epidemiology ; virology ; Disease Outbreaks ; Encephalitis ; epidemiology ; virology ; Enterovirus ; genetics ; Enterovirus B, Human ; genetics ; Female ; Genotype ; Humans ; Male ; Molecular Sequence Data

OBJECTIVETo study the metastasis and micrometastasis in No.14v lymph nodes in patients with lower third gastric cancer.

METHODSA retrospective study was performed. A total of 53 patients undergoing radical resections by a single surgeon for lower third gastric cancer in the Department of General Surgery at the Affiliated Hospital of Qingdao Medical College were included. Conventional pathological section was used to detect lymph nodes metastasis and telomere TRAP-ELISA was used to identify the micrometastasis in No.14v lymph nodes.

RESULTSA total of 96 lymph nodes were dissected from the No.14v group and lymph nodes metastasis were discovered in 9 patients by conventional pathological section. Forty-four patients had no metastasis on conventional pathological examination, of whom 13(29.6%) were found to have micrometastasis. The overall metastatic rate was 41.5%(22/53). Metastasis and micrometastasis in the No.14v lymph nodes were associated with Borrmann types, depth of invasion, No.6 lymph nodes metastasis, tumor diameter, and TNM staging(P<0.05).

CONCLUSIONSNo.14v lymph nodes in patients with lower third gastric cancer is associated with a high incidence of metastasis and micrometastasis. The status of No.6 lymph nodes may be used as an useful indicator for No.14v lymph nodes metastases during the operation.

Adult ; Aged ; Female ; Humans ; Lymph Node Excision ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; pathology ; Male ; Middle Aged ; Neoplasm Micrometastasis ; pathology ; Retrospective Studies ; Stomach Neoplasms ; pathology ; surgery