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.

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).

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

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.

Objective To compare the effect of low concentration versus ultra-low concentration of ropivacaine in combination with sufentanil delivered via programmed intermittent epidural bolus (PIEB) with patient-controlled epidural analgesia (PCEA) during labour on obstetric and anesthetic outcomes.Methods Seventy-six ASA Ⅰ or Ⅱ nulliparous parturients who were at full term with a singleton fetus in vertex presentation were randomized to receive 0.1 ％ ropivacaine with 0.5 μg/mL sufentanil (Group L,38 cases) or 0.06％ ropivacaine with 0.5 μg/mL sufentanil (Group UL,38 cases).Another matched 76 cases of primiparas without labor analgesia (Group C) served as controls.For primiparas received analgesia,an epidural catheter was inserted when the cervical dilatation was up to 2-3 cm.The analgesia level was controlled below T8,and VAS score was controlled below 4.Thirty minutes after analgesia started,Group L and UL received PIEB with PCEA regimen including basal infusion of 8 mL/h of ropivacaine with sufentanil,patient-controlled bolus 6 mL and lockout interval 10 minutes.The duration of labor,delivery mode,Apgar score,VAS score and anesthetic drug consumption of each group were recorded.Results The second stage of labor were not statistically different between Group UL and C.The second stage of Group L was longer than that of Group C (P＜0.05).Furthermore,with similar performance in pain score and satisfactory level at each time point,Group UL consumed much less ropivacaine than Group L (P＜0.05).The cesarean section rate,instrumental delivery rate and the Apgar score were not significantly different among the 3 groups.Conclusions While using PIEB with PCEA,0.06％ ropivacaine was capable of providing satisfactory analgesia effects for primipara and has less effects on the obstetric outcome.Therefore,it could be recommended for labour analgesia.

Objective:To study heat shock proteins(HSP) and their association with Hantavirus nucleocapsid protein(HV-NP) in Vero-E6 cells infected with Hantaan76-118.Methods: The HV-NP was identified by immunocytochemical staining after infected with Hantaan76-118.The expression of HSP was detected by Western-blot and analyzed by double specific antibody sandwich ELISA.Results: Western-blot exhibited that the expressions of HSP27,HSP70 and Grp94 in the Vero-E6 cells infected with Hantaan 76-118 were significantly higher than those in the control cells(P

Objective:To study the law and significance of stress response after Vero-E6 cells infected with Hantavirus (HTV). Methods:The law of expression of HSP 70 and its mRNA level were detected by immunocytochemical staining, laser confocal microscope and RT-PCR. Results:After being infected with HTV, immunocytochemical staining exhibited that the expression of HSP 70 increased. laser confocal microscope and RT-PCR detection showed that the level of HSP 70 and its mRNA increased quickly and kept at a high level in comparison with the normal control(P