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.

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 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

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.

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 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

OBJECTIVETo assess the association between single nucleotide polymorphisms (SNPs) of calcium channel β 2 subunit (CACNB2) gene and essential hypertension (EH) in ethnic Han Chinese in Wenzhou area, and to study the influence of rs7069292 alleles on gene expression with luciferase reporter technique.

METHODSSixty hundred and thirty seven Han Chinese with EH and 600 normal controls were enrolled. Genotypes of 6 SNP within CACNB2 gene including rs2228645, rs2357928, rs7069292, rs7099380, rs10764319 and rs11014166 were determined with matrix assisted laser desorption ionization/time of flight mass spectrometer (MALDI-TOF MS). A luciferase reporter gene plasmid containing the fragment flanking rs7069292 (-2831 bp to -2460 bp) in the 5' regulatory region of CACNB2 was constructed.

RESULTSCompared with the control, CT and TT genotypes for the rs7069292 locus were significantly more common in EH group (5.20% vs. 2.17%, 2.59% vs. 1.08%, P< 0.05). CC genotype was not found. Promoter activity for allele C of the rs7069292 locus was significantly increased compared with allele T (P< 0.05). No significant difference was detected for other 5 SNPs in terms of genotypes and allele frequency.

CONCLUSIONThe rs7069292 CT polymorphism of the CACNB2 gene is associated with EH in ethnic Han Chinese from Wenzhou area. A T>C mutation may affect the expression of CACNB2.

Aged ; Alleles ; Base Sequence ; Calcium Channels, L-Type ; genetics ; Case-Control Studies ; Cell Line ; Female ; Gene Frequency ; Genetic Predisposition to Disease ; Genotype ; Humans ; Hypertension ; genetics ; Male ; Middle Aged ; Polymorphism, Single Nucleotide

Objective To explore the effect of Internet addiction on adolescent's attention.Methods Neuropsychological evaluations of attention function were done separately in 18 adolescents who met the diagnostic criterion for Internet addiction and in other 18 ones without Intemet addition tendency as a control group. With auditory and visual oddball paradigms, and stimulated by standard,target and novel stimuli, EEG was recorded and analyzed to get the event-related potential P300 and compare the latency and amplitude of P3a and P3b between the 2 groups. Results Compared with the controls, the attention of Internet addiction group was decreased significantly in neuropsychological evaluations. The latency of P3a potentials induced by novel stimulus was much shorter and the amplitude of it was higher in addiction group than in control group, but the latency of P3b potentials generated by target stimulus was prolonged and the amplitude decreased obviously. Conclusions Intemet addiction can cause damage to adolescent's attention function. There is a correlation between the attention impairment and the change of P300 potentials.