Epigenetic regulation involved in eukaryotic gene expression plays an important role in the progression of colorectal cancer. Colorectal cancer epigenetics is evolved in DNA methylation which is associated with the activation of oncogene and inactivation of tumor suppressor gene. The potential reversibility of DNA methylation offers exciting opportunities for therapy and diagnosis of colorectal carcinoma.

Objective To establish a rapid, sensitive, and specific quantitative method to detect hepatitis C virus. Methods A primer set targeting HCV 5'UTR was designed. The isothermal amplification was performed by the Bst DNA polymerase and AMV reverse transcriptase, under the temperature of 60℃ for 60 min. The signal was monitored by SYBR Green Ⅰ. Results One hundred and twenty positive serum samples, confirmed by the real-time PCR. All were detected by the isothermal amplification, while 110 healthy subjects' samples were negative by the both methods. The lower detect limit was determined to 10 IU/ml HCV-RNA, by the assay on serial dilutions of the quality control standards obtained from clinical investigation center of MOH. Conclusion A real time reverse loop-mediated isothermal amplification method was developed to detect HCV, with the characteristic of rapidity, high sensitivity and specificity.

Objective To summarize clinical manifestations and diagnostic criteria of ischemic reperfusion (IR) injury to the spinal cord. Method The clinical manifestations and management of 15 cases of spinal cord IR injury after operation during 2000 to 2005 were retrospectively analyzed. Result All the patients with spinal cord IR injury presented progressive ascending motor and sensory functional impairment beginning from lower extremities 3h post operation, and they were all treated immediately with methylprednisolone and neurotrophy drugs to restore spinal cord function. However, mechanical compression with organic lesion should be ruled out to establish the diagnosis of spinal cord IR injury in the retrospective analysis. Different stages of spinal cord IR injury occurred in these 15 cases. Through symptomatic treatment, 8 patients recovered completely; the clinical symptoms were basically improved in 4 cases who could lead a normal life; in 3 patients clinical symptoms were improved but not satisfactory. Conclusion Though spinal cord IR injury seldom occurs in the clinic, its damage is disastrous. Proper management is critical to save patients from poor life quality.

Objective A retrospective study of the results of surgical or non-operative treatment in patients with herniation of lumbar disc 10 years or longer before,in order to evaluate the efficacy of different therapeatic strategies for lumbar disc herniation. Method Definite diagnosis of lumbar disc herniation was made in all cases before treatment,and the follow-up period was 10 to 38 years. The patients were divided into three groups according to different treatment methods: non-operative group,simple nucleus resection group,and nucleus resection combined with space fusion group. The long term therapeutic effects and image change of the three groups were observed. The difference of height of intervertebral space at the herniation site was measured before treatment and at the latest follow-up. Adjacent spaces were also observed. The data were analyzed statistically by t -test and ? 2 -test. Results Among 98 cases in non-operative group,85 cases enjoyed almost a normal life and work;vertebral canal decompression was performed for the rest 13 cases because of intermittent claudication. The protruded space was narrowed for 3.17?1.65mm in height in average after non-operative treatment,and there was degeneration of adjacent spaces in 13 cases. Among the 84 cases in simple nucleus resection group,relative good outcome was seen in 71 patients,and the remaining 13 cases had to receive a second operation because of symptom relapse. The protruded space was decreased 5.50?1.51mm in height in average as measured at the latest follow-up comparing with that of before operation. The degeneration of the adjacent space was observed in 15 cases. In 71 patients who received nucleus resection combined with space fusion experienced a good relief of symptoms. The space,where disc herniation had occurred,fused very well and no re-narrowing occurred. Degeneration of the adjacent space was observed in 39 cases,but the patients were symptomless. Conclusions The effect of either non-operative therapy or operation for patients with lumbar disc herniation was good after a long-term follow-up if the indication was chosen properly. We suggest that non-operative therapy should be the first choice for the lumbar disc herniation. The relatively normal nucleus should be preserved as much as possible when nucleus resection was performed. Intervertebral fusion was a relative better treatment comparing with the other two treatments.

Objective To investigate the HAART in PMTCT different intervention time of blocking effect. Methods 49 cases who received out -patient examination and hospital delivery of HIV positive pregnant women were selected,the intervention time pregnant 14 weeks,pregnant 28 weeks.The patients were divided into three groups,group A (n =20),group B (n =19),group C (n =10).The safe delivery,the newborn 50 out of 18 months of tracking and detection,and the results were analyzed.Results Three groups of antiviral HIV infected maternal immune status (P >0.05 ),antiviral CD +4 cell number variation had no statistical significance (P >0.05 ),the blocking effect was A group of infants 0 infection.In group B,1 cases of infant infection.Group C 2 cases of infant infection.Compared between group A and group B,the difference was not statistically significant (χ2 =0.97,P >0.05),compared between group B and group C,the difference was not statistically significant (χ2 =1.40,P >0.05). Compared between group A and group C,the difference had statistical significance (χ2 =4.08,P <0.05 ). Conclusion HAART for blocking HIV mother to child transmission effect is very significant,if in 14 weeks pregnant initiation of HAART,the infant HIV infection rate is lower,recommended the early antiviral treatment and after childbirth to strengthen follow -up and management.

Objective To study the expression of connexin (Cx) genes and the effects of a variety of inducers on the expression changes of Cx and mutations of Cx coding sequences in human gastric cancer. Methods Northern Blot, RT PCR and PCR single strand conformational polymorphism were used. Results 1.There were regular expression patterns of Cx genes in human normal gastric epithelium, paracancerous tissues and gastric cancers. 2.RA and DMSO can induce the expression of Cx43 in gastric cancer and decrease of Cx46 expression by RA and TPA induction. 3.There was no any mutation in Cx43 coding sequences. Conclusion 1.Cx32 gene may be a specific gene in genome of gastric epithelium constructing gap junctional intercellular communication. 2.Cx43 is a inducible gene in gastric cancer cells. 3.The down regulation of Cx43 expression is not caused by mutation of Cx coding sequences in human gastric cancers. 4.The authors proposed a hypothesis on the variation of expression of Cx genes in human gastric cancer.

In four patients with chronic pancreatitis from two hereditary pancreatitis (HP) families and 63 normal controls, five exons of cationic trypsinogen gene (PRSS1) were amplified by PCR and it's products were analyzed by sequencing, related clinical data were also collected. All the four patients were found mutations in the PRSS1 gene but their clinical feature is absolutely different. Six patients with diabetes mellitus were found in pedigree No. 1, it's members show pancreatitis symptom later, at about 29, the tumor markers (CA19-9, CA72-4) is obviously higher than the patients in pedigree No. 2, two patients with chronic pancreatitis in pedigree No. 2, show symptom earlier without diabetes mellitus, their clinical characterization are different too. The number of CD4+T cell/CD8+T is very low in Ⅲ 8, but Ⅲ 7 is normal, and the level of anti-HBs of Ⅲ 8 is variable in the course of pancreatitis, but the phenomenon was not found in Ⅲ 7. In their PRSS1 gene two guanosine (G) to adenosine (A) mutations were found in PRSS1 exon 3 of pedigree No. 1, one was detected at 336 basyl, the other mutation occurs at 361 basyl. The results of the mutations were Lys →Lys and Ala →Thr. While thymine (T) to adenosine (A) and (guanosine) G→(adenosine) A mutation in PRSS1 exon 3 was detected in the other patient of pedigree No. 2 (Ⅲ 8). One was 361 basyl, the other at 415 basyl. While c.415 T→A was not found in the proband of pedigree No. 2 PRSS1 gene (Ⅲ 7). All of the mutations were heterozygous mutation, that is to say all of the trypsinogen were wild type and mutant type concomitance, the normal and abnormal pathway of active trypsinogen exist partially. At the same time, the mutations of SPINK1 were not observed. Compared with the documents and registration of NCBI, it can be concluded that PRSS1 gene had many kinds of mutations in hereditary pancreatitis, the heterozygous mutations (c.336 G→A, c.415 T→A) were the novel mutations and related with clinical phenotype. What's more, it's the first time that the multisite heterozygous mutations of PRSS1 gene were reported. The presence of the mutations in four patients with chronic pancreatitis, it's absence in their relatives and the strong evolutionary conservation of the mutation, all indicate that the trypsinogen mutation is associated with hereditary pancreatitis and for the first time raises the question whether a gain or a loss of trypsin function participates in the onset of Chinese pancreatitis.

This paper introduces the structure, contents, function of the digital teaching re-courses database in Clinical Skills course of Central South University, analyses the essence and char-acter of the flipped classroom (FC), and emphasizes flexible environment, learning culture, inten-tional content, professional educator in its implementation process. Meanwhile, it introduces the appli-cation of flipped classroom based on digital resources database in the teaching of Clinical Skills course, mentions the importance of the four aspects:preparing teaching materials, learning before class, guidance and cooperation in class, evaluation and knowledge expansion after class. Lastly, it analyses the advantages and foreground of it.

Objective: To evaluate the effect of NGX6 with 5-Fu on the apoptosis of colon cancer cells. Methods: The NGX6-transfected HT-29 cell line with 5-Fu was used in the test group. HT-29 cell line with 5-Fu and PDTC was used in the control group. The expression of NF-κB was detected by EMSA. The proliferation of HT-29 cell line was assayed by MTT. The effect of NGX6 on the apoptosis was detected by FCM. HT-29 cells were double-stained by PI/Annexin-V and AO/EB and observed by fluorescence microscopy. Results: The expression of NF-κB was inhibited in NGX6 transfected colon carcinoma cell group and in colon carcino-ma cell group treated with PDTC. Treatment with the chemopreventive compounds 5-Fu and PDTC resulted in different responses in the effects of anti-proliferation and induced apoptosis of colon carcinoma cells. There was no significant difference in apoptosis between NGX6-transfected HT-29 call line with 5-Fu and the cells in the control group. NGX6 gene enhanced the effect of 5-Fu on the proliferation and apoptosis of colon carcino-ma cells. Conclusion: NGX6 gene can induce apoptosis and inhibit the proliferation of colon carcinoma cells. NGX6 gene can enhance the effect of 5-Fu on the proliferation and apoptosis of colon carcinoma cells through NF-κB pathway.

Objective To investigate the infection status of nanobacteria on patients of chronic hepatopathy and hepatocellular carcinoma,and evaluate the clinical value of PCR.Methods In sera of 68 cases of chronic hepatitis B(CHB),56 chronic severe hepatitis B(CSHB),66 cirrhosis of liver(CL)and 23 hepatocellular carcinoma(HCC),nanobacteria were detected by immunohistochemistry stain(IHC),Transmission Electron Microscopy(TEM)and polymerase chain reaction(PCR),compared with 40 healthy people.Results The positive rates of PCR were 27.69%,50.00%,61.29%,52.38% and 5.00% in patients with CHB,CSHB,CL,HCC and normal control respectively(P