The efficient characterization of genetic and epigenetic alterations in oncology requires highly sensitive and specific high throughput procedures.However, the necessary level of sensitivity and specificity were previously unreachable using conventional testing procedures.By partitioning individual target molecules within separated compartments, digital PCR ( dPCR ) could allow to overcome such limitations and detect with unprecedented accuracy very rare sequences.In such procedure, the sample is diluted such that each individual compartment will contain no more than one target sequences.The assay provides an absolute value and quantitative data.The recent coupling of dPCR procedure with microfluidic systems in commercial platforms should make droplet based digital PCR an essential tool for the management of patients with cancer.Applications range from the analysis of tumor heterogeneity to the analysis of body effluents.Droplet based digital PCR is also particularly suited for the increasing field of liquid biopsy analysis.

Objective To develop a novel targeted delivery system for the treatment of brain metastases from breast cancer.Methods Micelles were prepared by membrane hydration method,and the micelles were connected to Angiopep-2.Results The micelles were confirmed to be well targeted in vitro.Conclusion Nano-particles with the targeting molecule Angiopep-2 has good brain-targeting function.

LSAB immunohistochemistry and digoxin-labeled in situ hybridization methods were used to detect the expression of EGFR and TGF-a and the transcription of EGFR-mRNA in human renal cell carcinoma (RCC) tissues. The expression rate of EGFR and TGF-α in 46 cases of human RCC tissues were significantly higher than that in 38 cases of corresponding autologous normal kidney tissues (EGFR.. 53. 4 % vs 21.0 %;TGF-α: 39. 1 /% vs 13. 2 %, P＜0. 05). Both EGFR and TGF-α were simultaneously overexpressed in some cases of RCC tissues. No relationship existed between EGFR or TGF-α and the RCC staging and grading. The positive rate of transcription EGFR-mRNA in 25 cases of RCC tissues was significantly higher than that in 20 cases of corresponding autologous normal kidney tissues (44 % vs 15 %, P＜0. 05). The above findings demonstrated that RCC tissues overexpressed EGFR and TGF-αand overtranscribed EGFR-mRNA. The overexpressed EGFR and TGF-α might contribute to the growth and development of RCC by taking part in the autocrine growth loop in RCC.

BACKGROUND:Al ogeneic penetrating keratoplasty is the most effective method for treating corneal blindness. However, the incidence of rejections is high after keratoplasty, so it is urgent to develop an immunosuppressive drug with high efficacy and low toxicity. OBJECTIVE:To establish al ogeneic penetrating keratoplasty models and monitor the expression of tumor necrosis factor-αin blank control group and after transforming growth factor-β1 eyedrop during acute rejection period of corneal grafts. METHODS:Al ogeneic penetrating keratoplasty models were established and were randomly divided into blank control group, ciclosporin A group (1%ciclosporin A), and transforming growth factor-β1 group (1μg/ml transforming growth factor-β1 eyedrop). The medications from each group commenced at 1 day after surgery, one eyedrop once, three eyedrops per day. Al the operated eyes were given 0.3%ofloxacin ophthalmic solutions and 0.5%tropicaide ophthalmic solution, three times per day, for 12 days. The corneal grafts were harvested for hematoxylin-eosin staining and immunihistochemical staining (SABC method), to detect tumor necrosis factor-αexpression in corneal grafts. RESULTS AND CONCLUSION:Hematoxylin-eosin staining showed that, corneal grafts were significantly thickened, a large number of histoleucocytes and lymphocytes infiltrated in the blank control group;corneal grafts showed normal thickness and no inflammatory cel s infiltrated in the transforming growth factor-β1 group. Immunohistochemical staining showed that, there were less cel s positive for tumor necrosis factor-αin the transforming growth factor-β1 group compared with the blank control group (P<0.05). Transforming growth factor-β1 eyedrops can reduce the expression of tumor necrosis factor-αin the corneal grafts during acute rejection period, and reduce the inflammatory cel s infiltration in the corneal grafts, which is probably the mechanism of transforming growth factor-β1 to prevent and treat corneal al ograft rejection.

Purpose:To evaluate radiotherapy after operation of esophageal carcinoma. Methods:A comparative study was done between the group of 80 patients treated by postoperative radiotherapy and the group of 80 patients by operation only from January 1989 to June 1994. The radiation dose was 40—50 Gy. Results:The 1 ,3 ,and 5 year survival rates of operative group were 76.3%, 37.5%, 22.5%, and those of postoperative radiotherapy were 77.5 %,56.3 %,32.5 %. There was significant difference between the 3 year survival rates of the two groups ( P

Objective To explore the pattern and quantify the heat shock protein HSP)70 and HSP70 mRNA in Vero-E6 cells after infection with Hantann virus(HTNV).Methods The expres- sion of HSP70 and change of its mRNA level were detected by immunocytochemical staining,nucleic acid hybridization in situ and RT-PCR.Results In situ hybridization and RT-PCR were used to eval- uate the level of HSP70 mRNA during Hantaan 76-118 infection.HSP70 mRNA increased 0.5 h after infection,reached its peak by 12 h and gradually declined to steady state level by 72 h(vs.sham infec- ted group,P＜0.05).The expression of HSP70 protein induced by Hantaan 76-118 infection was e- valuated by quantitative immunocytochemical staining.HSP70 increased 0.5 h after infection,reached its peak by 12 h and decreased at 72 h after infection(vs.sham infected group,P＜0.05).Conclu- sions HSP70 can be induced directly by HTNV infection at both mRNA and protein levels,It pro- vides a basis for the further study of the pathogenesis,prevention and treatment of hemorrhagic fever with renal syndrome(HFRS).

Objective To investigate outpatients' awareness of the essential medicine system and their intention to use it at county hospitals,analyze the problems in the implementation process and give suggestions.Methods Using questionnaires to survey randomly intercepted patients.Data so acquired were keyed in twice with Epidata 3.0 and SPSS 16.0 was used for analysis featuring descriptive statistics.Results Among the 1064 patients surveyed,only 159 (14.9％)have heard of essential medicines,and the top three ways of awareness were respectively from medical staff,television and Internet; patients pay most attention to drug efficacy in their daily life and the majority of them follow doctors' advices.Conclusion It is necessary to strengthen promotions,formulate diversified publicity strategy,train doctors to guide patients to use essential medicines rationally.

Background Previous study on critical period plasticity in local cortical circuits primarily was only to test the role of some proteins in visual cortex on visual development based on the existed neural signals expression system,but whole containing proteins analysis in cortical circuits is lack.To perform a whole containing proteins analysis has an important significance for critical period plasticity study.Objective This study was to investigate the influence of dark rearing on visual sense and proteomics in visual cortex for growing rat.Methods Two SD female rats were fed in two cages together with 12 newborn rats on the same day respectively,and half number of newborn rats were exchanged each other from first day after delivering and marked by eartipping.The newborn rats in a cage were bred in the dark environment for 40 days,and newborn rats in other cage were bred in the nature environment as controls.The blink response of rats to nearby object was examined and compared between the two groups of rats.Then three rats from two cages were sacrificed respectively and bilateral primary visual cortex tissue was isolated.Proteomics in rat primary visual cortex was detected by two-dimensional electrophoresis and mass spectrometry and the result was checked from database.Then the survival rats in the dark environment returned to the nature environment for 10 days,and the blink response of rats to nearby object were compared with that of agematched rats in the natural environment.The use and care of experimental rats followed the instruction of Ethic Committee of Nankai University.Results The blink response of rats was (0.33 ± 0.35) times in the dark environment for 40-day group,and that in the natural environment for 40-day group was (6.42±0.68) times,with a significant difference between them (t =24.38,P＜0.01).After returned to natural environment for 10 days,the blink response times of rats were less than those of the natural environment group ([5.00±1.22] times vs.[6.11±0.59]times),but this change was not statistically significant (t =2.09,P＞0.05).Two-dimensional electrophoresis and mass spectrometry assay revealed that 36 different proteins in visual cortex were found in the dark-feed rats compared with the natural environment rats,including 26 loss proteins and 10 extra proteins.Among the different proteins,Eps 15 homology domain-containing protein-3 (EHD3),tubulin alpha-1A chain and 2 ',3 '-cyclic-nucleotide 3 ' phosphordiesterase were the known proteins.Conclusions Dark rearing cause reversible visual loss in critical period plasticity newborn rat,and the change of proteomics in visual cortex is probably an affecting factor.

BACKGROUND:Colagen materials have good biocompatibility and biodegradability, but also had some problems such as low mechanical strength, poor resistance to degradation exposed in the process of clinical application. Numerous studies have reported that proper crosslinking could improve the disadvantage of colagen materials, regulate porous network structure, sweling and degradation of colagen materials. OBJECTIVE: To optimize carbodimide crosslinking process of colagen sponge and determine the best process conditions. METHODS:Colagen sponge was cross-linked by carbodimide for the preparation of loose and porous colagen sponge. Meanwhile, we optimized the conditions of cross-linking, in which the selected concentration of carbodimide was 5, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100 mmol/L, linking time was 2, 4, 6, 8, 12, 16, 20, 24 hours, and linking temperature was 5, 10, 15, 20, 25, 30, 35℃. We evaluated the best process conditions of colagen sponge through the aperture, porosity, water absorption, and degradation rate. RESULTS AND CONCLUSION:The optimal conditions were carbodimide concentration 50 mmol/L, crosslinking temperature 20℃,crosslinking time 6 hours. At this point, the average pore diameter of colagen sponge was 105 μm, the porosity was 79.45%, water absorption was 287.14%, and the degradation rate was 15.04% (2 days). The crosslinking of colagen sponge significantly improved its water absorption and degradation resistance.

This study aimed to summarize the diagnosis and treatment of uncommon complications in esophageal and cardiac cancer operations as well as obtain lessons from the failure. Methods:The etiology, clinical diagnosis, treatment, prevention measures, and factors that contribute to the failure of the uncommon complications were analyzed retrospectively. Results: Results showed ten cured cases and eight death cases (except for the reported eight cases). Conclusion:The following conclusions were ob-tained. 1) The complications described in this paper were rare, but they sometimes occur clinically. If wrong treatment was taken, seri-ous consequences would be expected. 2) Once a jet-like bleeding of aortic esophagus avulsion injury occurs, the surgeon must be calm and take proper treatment to successfully patch up the wounds. 3) The right thoracic approach is a new approach to prevent the injury of azygos vein. 4) Patients with anastomotic stoma fistula easily cause a delay in diagnosis if the leakage was wrapped in the inferior phrenic. 5) Taking preventive measures is the key method for the anastomotic aortoesophageal fistula. 6) Occurrence of pulmonary em-bolism after operation should be monitored. 7) Thoracic gastric mediastinal hernia would cause serious consequences if treatment was not taken at a proper time.