1.Clinical application of digital PCR for accurate detection of gene mutation in cancer patients
Chinese Journal of Laboratory Medicine 2016;39(3):150-153
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.
2.Brain Targeting Drug Nano Micelle in Vitro Experimental Study
Journal of Medical Research 2017;46(5):55-59
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.
3.The Distribution of EGFR and TGF-α in Human Renal Carcinoma Tissues and Its Implication
Heng LI ; Xulin XIONG ; Gongcheng LU
Acta Medicinae Universitatis Scientiae et Technologiae Huazhong 2000;29(5):420-422
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.
4.Postoperative irradiation after radical surgery of esophageal carcinoma
Lu CHAO ; Kaijiong SHI ; Heng WANG
China Oncology 1998;0(01):-
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
5.Effects of transforming growth factor-beta1 on tumor necrosis factor-alpha expression of corneal allografts during acute immunological rejection in rats
Heng WANG ; Xiaohe LU ; Wei ZUO
Chinese Journal of Tissue Engineering Research 2015;(5):778-782
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.
6.Quantitative analysis of the patterns of heat shock protein 70 expression induced by Hantaan virus infection in vitro
Lu YU ; Heng MA ; Chun-Guang DUAN ;
Chinese Journal of Infectious Diseases 2001;0(06):-
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).
7.Analysis on outpatients' awareness of the essential medicine system and their intention to use it in county hospitals
Shuman XU ; Hua LU ; Jingya WU ; Heng WANG
Chinese Journal of Hospital Administration 2013;29(7):492-495
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.
8.The Expression of HSP induced by Hantavirus in Vero-E6 cells in vitro
Lu YU ; Heng MA ; Chunguang DUAN ; Shoujing YANG
Journal of Medical Postgraduates 2003;0(09):-
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
9.The effect of low concentration vs.ultra-low concentration of ropivacaine delivered via intermittent epidural boluses for labour analgesia on obstetric and anesthetic outcomes
Heng LU ; Shaoqiang HUANG ; Jianying HU ; Liang JIN ; Dan XU
Fudan University Journal of Medical Sciences 2017;44(4):498-502
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.
10.Study on optimum technology for extracting total flavonoids from Acanthus illicifolius by orthogonal design
Hongyou HU ; Heng ZHU ; Zhaochao ZHANG ; Changyi LU
Chinese Journal of Marine Drugs 2001;0(05):-
Objective To study the optimum conditions for extracting total flavonoids from Acanthus illicifolius through the two extraction processes.Methods An orthogonal test of L_9 (3~4)was designed to select optimum operation conditions for extracting total flavonoids from Acanthus illicifolius through Soxhelt and ultrasonic extraction process.According to the output of total flavonoids under the different extraction conditions,which were selected as ethanol contents,extraction temperatures and extraction time for Soxhelt extraction process, and extraction time,ethanol contents and ratio of liquid to material for ultrasonic extraction process.Results The optimum conditions were established as follows:ethanol content 60%, extraction temperature 85℃and extraction time 2.5h in Soxhlet extraction process;Ethanol content 50%,extraction time 50min,ratio of liquid to material 40:1 in ultrasonic extraction process.The extraction temperature was the main factor on the Soxhlet extraction process, and the ethanol content on the ultrasonic extraction process.Conclusion The technology of ultrasonic extraction is better than that of Soxhlet extraction to extract the total flavonoids from Acanthus illicifolius.The max output of total flavonoid from Acanthus illicifolius can reach to 3.82%.