Molecular diagnostics of lung cancer and its research progress / 中国实用内科杂志

Molecular markers are research hotspot in the field of malignant tumor research, and have important clinical significance for early diagnosis, prognosis monitoring and treatment of tumors. The precise treatment of individualized patients through molecular diagnosis has gradually become the consensus of clinical treatment of tumors. This article introduces the research progress of molecular diagnostic techniques in lung cancer.

Improving the thermostability of Trichoderma reesei xylanase 2 by introducing disulfide bonds

Background: Xylanases are considered one of the most important enzymes in many industries. However, their low thermostability hampers their applications in feed pelleting, pulp bleaching, and so on. The main aim of this work was to improve the thermostability of Trichoderma ressei xylanase 2 (Xyn2) by introducing disulfide bonds between the N-terminal and α-helix and the ß-sheet core. Results: In this work, two disulfide bonds were separately introduced in the Xyn2 to connect the N-terminal and α-helix to the ß-sheet core of Xyn2. The two disulfide bonds were introduced by site-directed mutagenesis of the corresponding residues. The half-life of the mutants Xyn2C14­52 (disulfide bond between ß-sheets B2 and B3) and Xyn2C59­149 (disulfide bond between ß-sheets A5 and A6) at 60°C was improved by approximately 2.5- and 1.8-fold compared to that of the wild type Xyn2. In addition, the enzyme's resistance to alkali and acid was enhanced. Conclusion: Our results indicated that the connection of the N-terminal and α-helix to the ß-sheet core is due to the stable structure of the entire protein.

Fluid shear stress regulates epithelial-mesenchymal transition (EMT) in Hep2 cells / 医用生物力学

Objective To examine the effects of fluid shear stress (FSS) on epithelial-mesenchymal transition (EMT) in Hep2 cells. Methods Hep2 cells were exposed to 140 mPa FSS. The morphologic changes of Hep2 cells exposed to FSS at different durations were observed using inverted microscope. The migration ability of Hep2 cells after FSS loading was investigated using scratch wound assay. The distribution and expression of cytoskeleton protein F-actin were examined by confocal microscope. The expression of the EMT marker proteins were detected by Western blotting. Results Most of Hep2 cells changed their morphology from polygon to elongated spindle with well-organized F-actin under FSS. After removing FSS, Hep2 cells recovered their initial morphology with flat polygon. FSS regulated Hep2 cells to enhance their migration capacity in a time-dependent manner. FSS promoted the rearrangement of cytoskeletal protein F-actin，which enhanced the migration behavior of Hep2 cells. In addition, FSS induced a time regularity of expression of the EMT marker proteins in Hep2 cells. Conclusions FSS as an important physical factor can induce EMT in Hep2 cells.

Efficacy of levetiracetam combined with short-term clonazepam in treatment of electrical status epilepticus during sleep in children with benign childhood epilepsy with centrotemporal spikes / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the efficacy of levetiracetam (LEV) combined with short-term clonazepam (CZP) in the treatment of electrical status epilepticus during sleep (ESES) in children with benign childhood epilepsy with centrotemporal spikes (BECCT).</p><p><b>METHODS</b>Fifteen children (9 boys and 6 girls) diagnosed with BECCT with ESES, who had continuous spike-and-wave accounting for over 85% of the non-rapid eye movement sleep as monitored by 24-hours ambulatory EEG or 3-hours video EEG, were enrolled. The clinical manifestations and EEG characteristics of patients were retrospectively analyzed. These children received two months of CZP treatment in addition to oral LEV [20-40 mg/(kg·d)]. All patients were followed up for 6-18 months.</p><p><b>RESULTS</b>The 15 children were orally given LEV in the early stage, but showed no improvement when reexamined by EEG or had seizures during treatment. Then, they received LEV in combination with short-term CZP. Re-examinations at 1 and 6 months after treatment showed that 14 cases had significantly reduced discharge (only little discharge in the Rolandic area) or no discharge, as well as completely controlled seizure; one case had recurrent ESES and two epileptic seizures during follow-up. The recurrent case received the combination therapy again, and re-examinations 1 and 6 months later revealed normal EEG; no seizure occurred in the 8 months of follow-up.</p><p><b>CONCLUSIONS</b>LEV combined with short-term CZP is effective and has few side effects in treating ESES syndrome among children with BECCT.</p>

Copper ions stimulate the proliferation of hepatic stellate cells via oxygen stress in vitro / 华中科技大学学报（医学）（英德文版）

This study examined the effect of copper ions on the proliferation of hepatic stellate cells (HSCs) and the role of oxidative stress in this process in order to gain insight into the mechanism of hepatic fibrosis in Wilson's disease. LX-2 cells, a cell line of human HSCs, were cultured in vitro and treated with different agents including copper sulfate, N-acetyl cysteine (NAC) and buthionine sulfoximine (BSO) for different time. The proliferation of LX-2 cells was measured by non-radioactive cell proliferation assay. Real-time PCR and Western blotting were used to detect the mRNA and protein expression of platelet-derived growth factor receptor β subunit (PDGFβR), ELISA to determine the level of glutathione (GSH) and oxidized glutathione (GSSG), dichlorofluorescein assay to measure the level of reactive oxygen species (ROS), and lipid hydroperoxide assay to quantify the level of lipid peroxide (LPO). The results showed that copper sulfate over a certain concentration range could promote the proliferation of LX-2 cells in a time- and dose-dependent manner. The effect was most manifest when LX-2 cells were treated with copper sulfate at a concentration of 100 μmol/L for 24 h. Additionally, copper sulfate could dose-dependently increase the levels of ROS and LPO, and decrease the ratio of GSH/GSSG in LX-2 cells. The copper-induced increase in mRNA and protein expression of PDGFβR was significantly inhibited in LX-2 cells pre-treated with NAC, a precursor of GSH, and this phenomenon could be reversed by the intervention of BSO, an inhibitor of NAC. It was concluded that copper ions may directly stimulate the proliferation of HSCs via oxidative stress. Anti-oxidative stress therapies may help suppress the copper-induced activation and proliferation of HSCs.

Copper ions stimulate the proliferation of hepatic stellate cells via oxygen stress in vitro / 华中科技大学学报（医学）（英德文版）

This study examined the effect of copper ions on the proliferation of hepatic stellate cells (HSCs) and the role of oxidative stress in this process in order to gain insight into the mechanism of hepatic fibrosis in Wilson's disease. LX-2 cells, a cell line of human HSCs, were cultured in vitro and treated with different agents including copper sulfate, N-acetyl cysteine (NAC) and buthionine sulfoximine (BSO) for different time. The proliferation of LX-2 cells was measured by non-radioactive cell proliferation assay. Real-time PCR and Western blotting were used to detect the mRNA and protein expression of platelet-derived growth factor receptor β subunit (PDGFβR), ELISA to determine the level of glutathione (GSH) and oxidized glutathione (GSSG), dichlorofluorescein assay to measure the level of reactive oxygen species (ROS), and lipid hydroperoxide assay to quantify the level of lipid peroxide (LPO). The results showed that copper sulfate over a certain concentration range could promote the proliferation of LX-2 cells in a time- and dose-dependent manner. The effect was most manifest when LX-2 cells were treated with copper sulfate at a concentration of 100 μmol/L for 24 h. Additionally, copper sulfate could dose-dependently increase the levels of ROS and LPO, and decrease the ratio of GSH/GSSG in LX-2 cells. The copper-induced increase in mRNA and protein expression of PDGFβR was significantly inhibited in LX-2 cells pre-treated with NAC, a precursor of GSH, and this phenomenon could be reversed by the intervention of BSO, an inhibitor of NAC. It was concluded that copper ions may directly stimulate the proliferation of HSCs via oxidative stress. Anti-oxidative stress therapies may help suppress the copper-induced activation and proliferation of HSCs.

Kallikrein-binding protein promotes axonal regeneration and protect rat retinal ganglion cells following optic nerve injury / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the effect of kallikrein-binding protein (KPB) in protecting retinal ganglion cells (RGCs) and promoting axonal regeneration following optical nerve injury in rats.</p><p><b>METHODS</b>Crush injury of the optic nerve at 0.5-1.0 mm from the eyeball was induced in rats, which received subsequent KBP injection into the vitreous cavity (experimental group) and PBS injection (control group). At 7, 14 and 21 days after the injury, the rats were sacrificed and frozen sections of the eyeball were prepared to observe the structure and thickness of the retina and count the number of survival RGCs with HE staining. The optic nerves were collected for Western blotting to assess the effect of KBP on the RGCs and axonal regeneration.</p><p><b>RESULTS</b>RGC counts and retinal thickness showed significant differences between the two groups. Western blotting also demonstrated a significant difference in the expression of the nerve regeneration marker protein GAP-43 between the two groups.</p><p><b>CONCLUSION</b>KBP offers protection on RGCs and promotes regeneration of the optic nerve axons after optic nerve injury in rats.</p>

Changes in glial fibrillary acidic protein and growth-associated protein-43 expressions in retinal ganglial cells during axonal regeneration / 南方医科大学学报

<p><b>OBJECTIVE</b>To explore the changes in the expressions of glial fibrillary acidic protein (GFAP) and growth- associated protein-43 (GAP-43) in retinal ganglial cells after neural transplantation.</p><p><b>METHODS</b>Thirty-nine rats were randomized into normal control group, nerve amputation group and nerve amputation with peripheral nerve transplantation group. Immunohistochemistry was used to detect the changes in the expressions of GFAP and GAP-43 at different time points after the operations, and real-time PCR was employed to detect the mRNA expressions of 13 genes in the retinal ganglial cells of the rats.</p><p><b>RESULTS</b>Immunohistochemistry showed obviously increased GFAP expressions in the retina following the nerve amputation. GFAP expression was down-regulated while GAP-43 expression upregulated in the retinal ganglial cells after peripheral nerve transplantation. Real-time PCR results showed that 5 days after the operations, retinal GFAP and GAP-43 expressions increased significantly in the nerve amputation group and peripheral nerve transplantation groups as compared with those in the control group, but GAP-43 expression decreased significantly in the former two groups afterwards.</p><p><b>CONCLUSION</b>The regenerated retina may adjust the production of GFAP. The retinal ganglial cells express GAP-43 during retinal regeneration. Up-regulation of the expression of GAP-43 provides the evidence for nerve regeneration following the nerve transplantation.</p>

Studies on spermatogonial stem cells cultured in vitro of Wuzhishan Mini Porcine / 生物工程学报

The isolation and culturation of SSCs of different stage of Wuzhishan Mini Porcine (WZSP) with different way of enzymatic digestion and culturation were deaded in this study. The results of the experiment described are as the following: The proper time of isolation and culturation of SSCs of WZSP is 1-20 old days. Different old of piglets with different method. Using DMEM medium as a fundmental culture medium add different gradient at 34 degrees C in a water-saturated atmosphere of 95% air, 5% CO2. The mulberry-shaped SSCs clusters appeared as original generation in 7-8 days culture. The SSCs clusters developed half-suspendedly in the culture medium. SSCs alkaline phosphatase (AKP) staining expressed positively. Mouse embryonic fibroblast was used as feeder layer for the SSCs passage cultured, The SSCs show good attached attributes, but the number of SSCs decreased quickly after 4 days culture. By seminiferous cord fragment culturation can also appear SSCs clusters in 5 days, The SSCs clusters developed half-suspendedly in the culture medium. In addition, the testes placed in cold (4 degrees C) PBS banlanced salt solution for 24 h also can be used as a good matierials for preparation of SSCs. These results indicate that the method of solation and culturation of SSCs are very correct and efficient, all these can be utilized as a good reference for future studies.

Comparison of retrograde amnesia changes within different injury levels of cerebral concussion in rats / 法医学杂志

OBJECTIVE@#To investigate the retrograde amnesia changes within different injury levels of cerebral concussion in rats.@*METHODS@#A metallic pendulum striker device of brain injury was deployed to duplicate CC models of different injury levels within Sprague-Dawley (S-D) rats. The investigated animals were divided into two groups according to classification standard, that is, Pure Cerebral Concussion (PCC) group and Complicated Cerebral Concussion (CCC) group. One control group was used, and each group included 8 animals. The retrograde amnesia of each group was assessed by Morris Water Maze (MWM) Test from 3 days preinjury to 7 days postconcussion.@*RESULTS@#Compared with the control group, the retrograde amnesia was detected within 3 days in PCC group, and 5 days in CCC group after injury. At the same time, the two groups both manifested space recognition deficit.@*CONCLUSION@#The retrograde amnesia existed in both pure cerebral concussion group and complicated cerebral concussion. Furthermore, the lasting time of retrograde amnesia in animals correlates to the injury level of brain concussion.

Establishment of maca mulatta model of resuscitation after selective cerebral ultra-deep hypothermic blood flow occlusion / 中华外科杂志

<p><b>OBJECTIVE</b>To study the feasibility of resuscitation after selective cerebral ultra-deep hypothermia and blood flow occlusion.</p><p><b>METHODS</b>Ten 4-10 year-old maca mulattas were divided into 3 groups: four-vessel occlusion group, two-vessel occlusion group and identical temperature perfusion group. MRI were examinated before and after operation, the vital signs and the hemodynamical parameters were observed during the experiment, neurological deficient evaluation was performed after operation.</p><p><b>RESULTS</b>In all of the ten monkeys, the hemodynamical parameters of two-vessel occulation were steady during the operation, and all of them lived after filling 60 minutes. MRI were normal after operation, and the function of neurological deficient scale was normal. The others of identical temperature perfusion group and four-vessel occlusion group were not resuscitation after filling 60 minutes and died.</p><p><b>CONCLUSION</b>Monkey could resuscitate from selective cerebral ultra-deep hypothermia and blood flow occlusion of bilateral common carotid artery in 60 minutes.</p>

Research of ultra-structural pathological changes of nervous, endocrine and immune system in heroin addicts / 法医学杂志

OBJECTIVE@#To investigate ultrastructural pathological changes of Heroin-Addicts.@*METHODS@#Heroin-Addicts' central nervous system, endocrine system, immune system and reproductive system in 4 cases are observed by using transmission electron microscope(TEM).@*RESULTS@#The changes of central nervous system are mitochondrion swelling, crista fragmentation and disappear. Endoplasmic reticulum dilation, nervous fibres and cell organelles reduction; mitochondrion swelling, Partial crista fragmentation and endoplasmic reticulum dilation are also found in endocrine system; Lymphocytes reduction, cytoplasm ingredient reduction and dead lymphocytes increase in immune system; in reproductive system, spermatogenic cells and cell organelles are reduced in the male and follicle disappeared in the female.@*CONCLUSION@#Ultra-structural pathological changes of heroin-addicts are presented acute, chronic oxygen deficiency degeneration and necrosis.