1.Construction of recombinant eukaryotic expression vector pEGFPCI-HIF-1α and its expression in eukaryotic cells PC12
Ninghui ZHAO ; Ran LI ; Xiaowei HUANG
Chinese Journal of Trauma 2010;26(2):160-164
Objective To construct the eukaryotic expression vector with the mutant of hypoxia inducible factor 1α (HIF-1α) that could over express nonhypoxic dependent HIF-1α and investigate its expression in eukaryotic cells PC12. Methods The coding sequence of HIF-1α was amplified from total RNA from the injured spinal cord of rats by means of RT-PCR. Then, overlapping extension PCR was employed to obtain one complete sequence encoding HIF-1α mutant without oxygen-sensitive degradation domain (ODD). By using gene recombination technique, the gene HIF-1α△ODD was inserted into eukaryotic expression vector _pEGFPC1 that contained a reporter gene of enhanced green fluorescent protein. The recombinant plasmid was successfully constructed and transfected into the neural cell line PC12. The expression of HIF-1α△ODD was analyzed by fluorescent observation and Western blot. Results A 2kb gene fragment encoding H1F-1α△ODD was successfully obtained by overlapping extension PCR. On this basis, the recombinant plasmid _pEGFPC1-HIF-1 α△ODD was constructed and testified with restriction enzyme digestion and DNA sequencing. After this recombinant plasmid was transfected into the neural cell line PCI2, both fluorescent observation and Western blot demonstrated that HIF-1α△ODD was over expressed under nonhypoxia conditions. Conclusion The eukaryotic expression vector _pEGFPC1-HIF-1α△ODD is successfully constructed and HIF-1α△ODD over expresses in transfected PCI2 cell lines.
2.Expression of hypoxia inducible factor-1? in spinal cord injury
Ninghui ZHAO ; Boyong MAO ; Wangning ZHOU
Chinese Journal of Trauma 2003;0(09):-
Objective To explore the expression pattern and effects of hypoxia inducible factor-1? (HIF-1?) in experimental spinal cord injury. Methods The expression of HIF-1? at various time was detected at levels of mRNA and protein by using methods of reverse transcription-polymerase chain reaction (RT-PCR),in situ hybridization (ISH) and immunohistochemistry. Results HIF-1? expressed more significantly at levels of mRNA and protein in all kinds of cells in the injured spinal cord than in the normal spinal cord (P
3.Influence of 256-slice Spiral CT Perfusion Imaging on the Cerebral Blood Flow before and after Cranioplasty
Jianping ZHU ; Ninghui ZHAO ; Zhi YAN ; Qiang YANG ; Xin SHI
Journal of Kunming Medical University 2016;37(7):48-52
Objective To evaluate changes in cerebral blood flow before and after cranioplasty by 256-slice Spiral CT perfusion imaging,and evaluate the effect of cranioplasty on the cerebral blood flow in patients with skull defect.Methods 256-slice spiral CT scan was performed in 20 cases with early cranioplasty surgery,CTP check time points were 1 to 2 days before and 10 to 14 days after cranioplasty surgery.We recorded the the CBF and CBV of the cortex,basal ganglia,and thalamus and other parts,MTT on rCBV,parameter values rCBF,MTT and 1TrP etc.and analyzed and compared.(RCBF,rCBV,MTT and TTP) Results The CBF of cortex after cranioplasty at injured side had statistically significant increase (P<0.05).The CBF of cortex,basic nuclei,thalamus on contrateral had no statistically significant difference.The cerebral blood flow on both sides of the basal ganglia and the thalamus was increased after surgery,but there was no significant difference between before and after surgery (P>0.05) Conclusion Cranioplasty can significantly improve the ipsilateral cortex cerebral blood flow,and CT brain perfusion can accurately assess changes in brain tissue blood flow before and after cranioplasty.
4.Analysis of Effectiveness of Deep Brain Stimulation for Forty Cases of Parkinson's Disease
Guoli SHI ; Mingrui ZHOU ; Jiachu ZHANG ; Ninghui ZHAO ; Boming SUN
Journal of Kunming Medical University 2016;37(9):113-115
Objective To investigate the effectiveness of deep brain stimulation (DBS) treating Parkinson's disease.Methods Forty cases of Parkinson's disease were selected from March 2014 to August 2015.The clinical symptoms of these patients were described and quantitatively analyzed with Unified Parkinson's Disease Rating Scale (UPDRS) before and after the procedure of DBS surgery.Results After deep brain stimulation surgery,the symptoms including muscle stiffness,static tremor,bradykinesia were improved,UPDRS scores were significantly lower and the demanding dosage of Parkinson disease drugs such as L-dopa/benserazide and L-dopa/carbidopa were also reduced.Conclusion Deep brain stimulation for Parkinson's disease is safe and effective.It can obviously control the symptoms,reduce the dosage of oral drugs,and improve the quality of life.
5.Decreased inhibitory synaptic function in CA1 region of hippocampus of adult rats following global ischemia-reperfusion
Lingfeng XU ; Wenpei MA ; Ninghui ZHAO ; Zhongtang FENG
Basic & Clinical Medicine 2006;0(02):-
Objective To observe the effects of global ischemia-reperfusion on inhibitory synaptic function in hippocampal CAl region of adult rats.Methods Animals were randomly divided into three groups: sham-operation group(SH),ischemia-reperfusion 3 day(IR-3) and 7 day group(IR-7).Global ischemic episode was achieved by 4-vessel occlusion.Evoked inhibitory postsynaptic currents(eIPSCs) were recorded from CA1 pyramidal cells in hippocampal slices using whole-cell voltage-clamp.Results The eIPSCs amplitudes generated by lower stimulus intensities were significantly decreased in both IR-3 and IR-7 rats as compared with SH rats.Moreover,the rise time of eIPSCs in IR-7 group was significantly decreased as compared with SH group(P
6.An approach for the measurement of the surface area of scalp flap over the cranial defect after decompressive craniectomy.
Qinhu ZHANG ; Lanjuan LIU ; Ninghui ZHAO ; Jinxi ZHAO ; Lian GAO ; He SUN ; Xinling SHI
Chinese Journal of Medical Instrumentation 2013;37(6):401-403
OBJECTIVETo introduce a simple, fast and universal measuring method used in measurement of the surface area of scalp flap over the cranial defect after decompressive craniectomy.
METHODSThe first step: CT images of the patient with craniocerebral trauma after decompressive craniectomy were obtained and imported into Mimics. The second step: based on the defined threshold, the 3D geometric models of brain and skull were reconstructed after the original Dicom format pictures three-dimensional processed by Mimics. The third step: based on the two builded 3D models, utilizing the segmentation and measurement tools of Mimics to conduct cutting, splitting and measuring operations for the 3D model of brain. The forth step: estimating the surface area of scalp flap over the removed bone flap by using mathematical computation methods.
RESULTSThe application of the introduced method estimated the surface area of scalp flap over the cranial defect of different people with different position of craniocerebral trauma.
CONCLUSIONSThis paper introduces a simple, fast and universal new measuring method. We can conveniently estimate the surface area of scalp flap by using the introduced method.
Decompressive Craniectomy ; methods ; Humans ; Imaging, Three-Dimensional ; Models, Anatomic ; Scalp ; surgery ; Surgical Flaps ; Tomography, X-Ray Computed
7.Comparison of Effects on Intraoperative Cerebral Metabolism between Total Intravenous Anesthesia and Intravenous Inhalation Anesthesia in Patients in Neurosurgery
Yijiao ZHOU ; Ping HU ; Shulan JIAO ; Ninghui ZHAO ; Luyan YANG ; Tianzi CHEN
Journal of Kunming Medical University 2016;37(12):103-106
Objective To compare the effect on intraoperative cerebral metabolism between the propofol combined with remifentanil infusion in total intravenous anesthesia and the desflurane combined with remifentanil intravenous inhalation anesthesia in patients in neurosurgery.Methods Thirty-four patients were randomly divided into the propofol combined with remifentanil group (group A,n=16) and the desflurane combined with remifentanil group (group B,n =18).The B-ultrasound under the guide of retrograde catheterization through right internal jugular vein and artery was prepared after the patients entered the operation room.Atropine,propofol,fentanyl,rocuronium were used in the induction of anesthesia.The mechanical ventilation was applied after conventional trachea cannula.Once patients were anesthetized steadily,jugular bulb venous oxygen saturation (SjvO2),jugular bulb venous oxygen partial pressure (PjvO2),Jugular Bulb venous hemoglogin (Hbv),jugular bulb venous lactate (Ljv),jugular bulb venous gluxose (Gv),arterial oxygen saturation (SaO2),arterial oxygen pressure (PaO2),arterin (Hba),arterial blood lactate (La),arterial blood gluxose (Ga),arterial blood oxygen content (CaO2),jugular bulb venous oxygen content (CjvO2),arteriovenous oxygen difference (AVDO2),cerebral extraction of oxygen (CEO2),cerebral lactate production rate (CLP) and cerebral glucose uptake rate (CGU) at different time [before anesthesia induction (T1),1 hours after the start of the operation (T2),2 hours after the start of the operation (T3),half an hour after the operation]were collected.Results (1) The value of SjvO2,PjvO2,CjvO2 and CaO2 in group B was significantly higher than that in group A (P<0.001);The value of AVDO2 and CEO2 in group B was lower than that in group A (P<0.05);(2) The value of Gv and Ga in group B was higher than that in group A (P<0.05);There were no significant differences about CGU in two groups (P>0.05);(3) The value of CLP in group B was lower than that in group A (P<0.05);there were no significant differences about Ljv and La in two groups (P>0.05);(4) Compared with the value at moment of T1 between group A and group B,the value ofPaO2,SaO2,PjvO2 and SjvO2 were increased with time (P<0.05),the value of CaO2,CjvO2,AVDO2 and CEO2 showed a downward trend (P <0.05).Conclusion (1) Both total intravenous anesthesia and intravenous inhalation anesthesia can reduce the cerebral oxygen metabolism;(2) For the cerebral protection of neurosurgery operation,it seems that the effect of intravenous inhalation anesthesia is more stronger than total intravenous anesthesia.
8.Research progress on"one-two punch"strategy in the therapy of glioma
Kui ZHANG ; Kai ZHAO ; Wenhu LI ; Minhao HUANG ; Ninghui ZHAO
Chinese Journal of Clinical Oncology 2023;50(21):1106-1109
Glioblastoma(GBM)originates from glial cells,and complete surgical resection followed by radiotherapy and chemotherapy is the current standard treatment.However,gliomas are subjected to not only accelerated cell death after radiotherapy and chemotherapy but also cellular senescence.Senescent cells produce a senescence-associated secretory phenotype(SASP),which has a dual effect on the tumor microenvironment.The"one-two punch"strategy of specifically eliminating senescent cells and inhibiting SASP-derived secretions provides a new direction for tumor therapy.In this article,we review the mechanisms that mediate tumor cellular senescence and SASP,the elimina-tion of senescent cells by senolytics for SASP inhibition,and the current situation of the"one-two punch"strategy for the treatment of glioma.