Effectiveness and safety of Jiejing Runmu decoction in treatment of dry eye disease.

OBJECTIVE: To investigate the effectiveness and safety of Jiejing Runmu decoction in relieving the clinical manifestations of dry eye disease (DED). DESIGN AND INTERVENTIONS: This single-arm prospective intervention study was conducted at the Peking University Third Hospital and People's Hospital of Yongqing. Of the 211 patients recruited, 200 completing the follow-up were included in the analysis. Patients received Jiejing Runmu decoction once a day for 4 weeks continuously, without any change in eye care habits. Individuals were evaluated at four time points: pretreatment (baseline), 2 weeks, 1 month, and 3 months (2 months after completion of treatment), using the Ocular Surface Disease Index (OSDI), tear film breakup time (TBUT), corneal fluorescein staining, Schirmer test I and meibomian gland assessments. Adverse effects were evaluated at each follow-up visit and systematic examinations were performed during the first and last visits. RESULTS: OSDI, TBUT, corneal fluorescein staining, Schirmer test I, meibomian gland expressibility, and quality of secretions improved at 2 weeks, 1 month and 3 months compared to baseline (P < 0.0001). No significant differences were found between the sexes. Patients above 45 years showed worse subjective symptoms and objective signs, and greater improvements in corneal fluorescein staining, meibomian gland expressibility, and quality of secretions were observed in this group. No obvious adverse effects were detected during any follow-up visit. CONCLUSION: Jiejing Runmu decoction significantly improved both the subjective symptoms and objective signs of DED, with favorable tolerance.

Genetic safety evaluation of allogeneic bone marrow mesenchymal stem cells in hosts following traumatic brain injury / 中华神经医学杂志

Objective:To investigate the genetic safety of allogeneic bone marrow mesenchymal stem cells (BMSCs) transplantation in traumatic brain injury (TBI).Methods:(1) In vivo experiment: BMSCs from male SD rats were isolated and cultured. Moderate TBI models were prepared by implanting and fixing micro-drug injection cannula into the left ventricle of 12 female SD rats, and 3 d after that, striking the right cerebral cortex of the rats with pneumatic precision percussion device was performed. Four h, and 3, 6, 9, and 12 d after modeling, TBI rats were given a single/multiple BMSCs infusion (2.5×10 5/time, total volume 10 μL) by cannula; 48 and 72 h, and 10 and 14 d after modeling, brain tissues of TBI rats (3 at each time point) were prepared into paraffin specimens. Immunofluorescent staining was used to detect the microglia activation, and RNAscope ? technology was used to detect the co-localization of astrocytes, neurons, microglia and transplanted BMSCs to observe whether the allogeneic BMSCs were integrated with the host brain cells after transplantation into TBI host. (2) In vitro experiment: the frozen and revived microglial cell line BV2 was transfected with green fluorescent protein (GFP)-positive lentiviral particles, and then, BMSCs prelabeled with pHrodo RED probe and BV2 cells pretreated with lipopolysaccharide were co-cultured in a certain ratio (BV2:BMSCs=1:1, 1:2, 2:1); after 36 and 72 h of co-culture, the phagocytosis between the 2 kinds of cells was observed under confocal fluorescence inverted microscope to observe the specific action forms of microglia on BMSCs. Results:(1) In vivo experiment: 48 and 72 h, and 10 and 14 d after modeling, no colocalization of transplanted BMSCs with astrocytes or neurons was found in paraffin sections of brain tissue in TBI rats; however, 10 and 14 d after modeling, microglia in TBI rats were obviously activated and migrated to the left lateral ventricle and choroid plexus, and co-localization of microglia with transplanted BMSCs was observed. (2) In vitro experiment: phagocytosis occurred after co-culture of BV2 cells at different proportions with BMSCs for 36 and 72 h. Conclusion:After transplantation, allogeneic BMSCs do not integrate with astrocytes or neurons of the TBI host, but they could be phagocytosed by microglia, indicating that allogeneic BMSCs transplantation for TBI is genetically safe.

Application of three-dimensional (3D) surgical display system in vitreoretinal surgery training / 中华医学教育探索杂志

At present, vitreoretinal surgery has gradually entered the era of minimally invasive surgery, and high-speed vitrectomy, micro-incision approach and wide-angle illumination have also pushed it to a higher level. Minimally invasive vitreoretinal surgery has become one of the key and difficult points in the teaching of ophthalmic microsurgery. The three-dimensional (3D) surgical display system can provide high-resolution, multi-level magnification, and stereoscopic images for surgery teachers and multiple observers at the same time, breaking the traditional "one-to-one" teaching of the main surgeon and assistant mirrors, realizing "one-to-more" and "head-up" surgery teaching, and thereby improving the teaching effect of vitreoretinal surgery.

Analysis of current ophthalmic microsurgery training system and exploration of hierarchical comprehensive training system / 中华医学教育探索杂志

In view of the problems and shortcomings of the domestic ophthalmic microsurgery training system, drawing lessons from the training programs of famous ophthalmic centers abroad, our hospital has explored a set of hierarchical comprehensive training system for ophthalmic microsurgery. Through the four levels-eight scales microsurgery training, the hierarchical comprehensive training system organically integrates the multimedia theoretical teaching, the microscopic practice of Wet-Lab laboratory, microscopic training of surgical simulator and the clinical practice to achieve a better teaching effect in clinical practice, being widely praised by teachers and students.

Correlation of NUF2 Overexpression with Poorer Patient Survival in Multiple Cancers / Journal of the Korean Cancer Association, 대한암학회지

Purpose@#NUF2 has been implicated in multiple cancers recently, suggesting NUF2 may play a role in the common tumorigenesis process. In this study, we aim to perform comprehensive meta-analysis of NUF2 expression in the cancer types included in the Cancer Genome Atlas (TCGA). @*Materials and Methods@#RNA-sequencing data in 31 cancer types in the TCGA data and 11 independent datasets were used to examine NUF2 expression. Silencing NUF2 using targeting shRNAs in hepatocellular carcinoma (HCC) cell lines was used to evaluate NUF2’s role in HCC in vitro and in vivo. @*Results@#NUF2 up-regulation is significantly observed in 23 out of the 31 cancer types in the TCGA datasets and validated in 13 major cancer types using 11 independent datasets. NUF2 overexpression was clinically important as high NUF2 was significantly associated with tumor stages in eight different cancers. High NUF2 was also associated with significantly poorer patient overall survival and disease-free survival in eight and six cancers, respectively. We proceeded to validate NUF2 overexpression and its negative association with overall survival at the protein level in an independent cohort of 40 HCC patients. Compared to the non-targeting controls, NUF2 knockdown cells showed significantly reduced ability to grow, migrate into a scratch wound and invade the 8 μm porous membrane in vitro. Moreover, NUF2 knockdown cells also formed significantly smaller tumors than control cells in mouse xenograft assays in vivo. @*Conclusion@#NUF2 up-regulation is a common feature of many cancers. The prognostic potential and functional impact of NUF2 up-regulation warrant further studies.

The effect of hormone therapy on the ocular surface and intraocular pressure for postmenopausal women: a systematic review and meta-analysis of randomized controlled trials.

OBJECTIVE: The aim of the study was to investigate the impact of hormone therapy (HT) on the ocular surface and intraocular pressure in postmenopausal women. METHODS: This systematic review and meta-analysis was conducted according to the Preferred Reporting Items for Systematic Reviews and Meta-analyses Statement. PubMed, EMBASE, Cochrane Library of Systematic Reviews, Cochrane Central Register of Controlled Trials, ClinicalTrials.gov, Chinese Biomedical Literature Database, and China National Knowledge Infrastructure were searched from inception to November 2019 without language restrictions. Only randomized controlled trials that evaluated the impact of HT on the ocular surface and intraocular pressure in postmenopausal women were eligible. The trials had to report at least one of the following outcomes: break-up time, Schirmer test, corneal staining, ocular surface symptom score, and intraocular pressure. Two investigators independently extracted the information, assessed the risk of bias, and evaluated the publication bias. All data were analyzed by Review Manager V.5.3. Sensitivity analysis and subgroup analysis were performed to find the source of heterogeneity and evaluate the different effects among subgroups. RESULTS: Nine randomized controlled trials (N = 612) were included. The HT group showed significant improvements compared with the control group in break-up time (mean difference [MD] = 2.09, 95% confidence interval [CI] 1.00-3.19, P = 0.0002), Schirmer test without anesthesia (MD = 4.17, 95% CI 1.55-6.80, P = 0.002), Schirmer test with anesthesia (MD = 1.44, 95% CI 0.71-2.18, P = 0.0001), and corneal staining scores (standardized mean difference [SMD] = -0.85, 95% CI -1.39 to -0.30, P = 0.002). Moreover, significant beneficial effects were observed on all four symptoms, including dryness (SMD = -1.21, 95% CI -1.99 to -0.44, P = 0.002), foreign body sensation (SMD = -1.02, 95% CI -1.29 to -0.76, P < 0.00001), ocular fatigue (SMD = -1.74, 95% CI -2.12 to -1.36, P < 0.00001), and burning (SMD = -0.53, 95% CI -0.78 to -0.29, P < 0.0001) after HT. Subgroup analysis revealed that, in terms of break-up time, postmenopausal women younger than 55 years achieved more improvements (MD = 0.88, 95% CI 0.16-1.59, P = 0.02) than women older than 55 years old (MD = 2.60, 95% CI -1.34 to 6.55, P = 0.20), and the estrogen subgroup received more benefits (MD = 3.11, 95% CI 0.93-5.30, P = 0.005) than the estrogen plus progestogen subgroup (MD = 0.42, 95% CI -0.02 to 0.85, P = 0.06). Sensitivity analysis and subgroup analysis suggested that the heterogeneity might derive from the methodological quality, the age of participants, and the intervention of the control group. Intraocular pressure (MD = -1.54, 95% CI -3.39 to 0.32, P = 0.10) was not evidently decreased after HT. No more specific adverse events (relative risk  = 1.66, 95% CI 0.41-6.77, P = 0.48) were found in the HT group. CONCLUSIONS: Our study revealed that HT could improve ocular surface function in postmenopausal women effectively and safely, especially for those who were younger than 55 years, and estrogen only showed more improvements than estrogen plus progestogen. The effectiveness of HT in treating dry eye in postmenopausal women is, however, still a controversial topic. In addition, we did not find HT led to a significant reduction of intraocular pressure.

Effect of human amniotic mesenchymal stem cell transplantation on motor behaviors in hSOD1-G93A mouse models of amyotrophic lateral sclerosis / 中华神经医学杂志

Objective To explore the therapeutic effect of intravenous transplantation of human amniotic mesenchymal stem cells (hAMSCs) on protection of motor behaviors in hSOD1-G93A mouse models of amyotrophic lateral sclerosis (ALS).Methods Amnion membranes were obtained from placentas delivered by healthy mother donors.The hAMSCs were gradually isolated and purified from amnion membranes using tissue culture method.Immunophenotypes of the isolated hAMSCs were analyzed using fluorescence activated cell sorter (FACS).Transgenic mice harboring a high copy number of hSOD1-G93A (B6SJL-TgN [SOD1-G93A] 1Gur) transgene were used in this study.Hemizygous transgenic progenies were maintained by mating the transgenic males with F1 hybrid wild-type (WT)females.The progenies were genotyped by polymerase chain reaction (PCR) using genomic DNA isolated from mouse tail after birth.The study included hSOD1-G93A mice transplanted with hAMSCs,PBS-injected transgenic mice,and normal WT mice (n=12).The hAMSCs were administered intravenously in jugular vein of the mice under anesthesia.The cells (1 ×106) in 200 μL PBS were delivered over 10 min.Animals received cells or PBS at 12,14,and 16 weeks old,respectively.The disease onset and progression of ALS mice models were monitored using rotarod performance test,PaGE test,and CatWalk gait analysis since 8 weeks old every week.Results (1) The immunophenotype of the isolated hAMSCs was conformed using FACS.These cells were positive for CD29,CD44,CD73,CD90,and CD166,but negative for CD14,CD34,CD45,CD123,and human leukocyte (site) DR antigen.Interestingly,stage specific embryo surface antigen 4 and octuber binding transcription factor 4 were detected in hAMSCs.(2) ALS mice in the hAMSCs transplantation group had significantly improved motor functions than those in the PBS treatment group:motor performance on the rotarod test (from 14 to 18 weeks old),PaGE test (from 15 to 18 weeks old) and CatWalk gait analysis (from 15 to 19weeks old) in hAMSCs-injected ALS mice was significantly improved as compared with that in the PBS treatment group (P＜0.05).Conclusions The multiple transplantation of hAMSCs by intravenous delivery can bring amelioration of the disease phenotype,as evidenced by improved motor function in hSOD1-G93A mouse models.The hAMSCs transplantation can be considered as a promising cellular treatment for ALS.

The role of vascular endothelial growth factor and bevacizumab on the wound healing in trabeculectomy patients / 中华实验眼科杂志

Proper wound healing and scaring are the key factor to the success of trabeculectomy in glaucoma patients.The inadequate wound healing will lead to bleb leakage and ocular hypotension after surgery;however,excessive wound healing and scaring will cause the failure of the surgery and eventually increase the intraocular pressure.The applying of antimetabolic drugs such as mitomycin C (MMC) and 5-fluorouracil (5-FU) are able to relieve the excessive wound healing in some degree;however,the side effects like ocular hypotension,dysesthesia endophthalmitis can never be ignored.What is worse,some patients are not sensitive to such drugs.Subconjunctival injection of CAT-152 (monoclonal antibody against transforming growth factor-β) was able to control wound healing in animal trabeculectomy model,while failed in multi-center clinical trial.Recent studies have focused on the role of vascular endothelial growth factor (VEGF) and VEGF inhibitors on the wound healing after trabeculectomy.This paper aims to review the mechanism of wound healing after trabeculectomy,as well as the role of anti-VEGF on this kind of wound healing and scaring.

Dynamic changes of Notch1 protein expression in the subventricular zone after traumatic cerebral injury of the rats / 中华神经医学杂志

Objective To observe the dynamic changes of Notch1 protein expression in the subventricular zone (SVZ) after traumatic cerebral injury (TBI) in rats,and to explore the correlation between Notch signaling pathway and neural stem cells (NSCs) proliferation after TBI.Methods Ninety-six SD rats were randomly divided into sham-operated group (only opening the scalp and shall window,n=16),control group (without any treatment,n=16) and experimental group (establishing medium-sized TBI models with Feeney's free fall method,n=64);rats in the experimental group were divided into 4 sub groups according to the execution times (one,three,7 and 14 d after TBI,n=16).Immunofluorescence staining was performed to detect the Notchl/nestin labeled cell expressions in SVZ of rats,and Notch1 protein expression was detected by Western blotting.Results Immunofluorescence results showed that the Notch1/nestin double staining positive cells in the right SVZ brain tissues of the experimental group on the first day of TBI had peak level,which was 2-3 times of the control group;and then,it gradually declined and basically returned to normal on 144 day of TBI;the differences between the 4 subgroups were statistically different (P＜0.05).The positive cell counts in the subgroups of first,third,and 74 d after TBI were statistically different as compared with those in the control group and sham operated group (P＜0.05).Western blotting results indicated that there was no significant difference in the Notch1 protein expression between control group and sham operated group;Notch1 protein expression in SVZ of the experimental group had peak level on the first day of TBI,gradual decline was noted after that,and normal level was noted on the 14th d of TBI.Conclusion TBI can activate the Notch1 protein expression,which induces the opening of Notch signaling pathway so as to affect the regulation of NSCs proliferation.

Expressions of cancer-related genes in human bone marrow-derived neural stem cells / 中华神经医学杂志

Objective To investigate the expression profile of cancer-related genes in human bone marrow-derived neural stem cells (Md-NSCs) to determine whether there are any characteristics that could help the evaluation of their tumorigenic potentials.Methods Md-NSCs were cultured in vitro and identified (experimental group);fresh human adult bone marrow cells were used as control group (sifting erythrocytes).The expression profiles of 440 cancer-related genes in cells from the two groups were analyzed by Oligo GEArray Human Cancer Microarray OHS-802;real-time quantitative PCR was performed to detect the expressions of oncogene MYC,matrix metalloproteinase 2 (MMP2),Notch congener 2 (Notch2),stanniocalcin 1 (STC1),integrin α3 (ITGA 3),signal transduction and transcriptional activation factor 5b (STA T5b),Ras congene gene family C (RhoC),and wingless-type MMTV integration site family member 1 (Wnt1).Results As compared with those in the control group,the Md-NSCs from experimental group had 66 tumor-related genes with high expressions (＞3 folds).MYC,MMP2,Notch2,STCI,ITGA3,STA T5b,RhoC and Wnt1 expressions in the Md-NSCs from experimental group were significantly higher than those in the control group (P＜0.05),whose results were accorded with genechip detection results,enjoying the folds of 4.35×100,2.84×100,2.87×100,3.41 ×102,2.22×102,6.99× 100,4.92 × 100 and 3.64 ×100,respectively.Conclusion A number of cancer-related genes are over-expressed in Md-NSCs,and the activations of some of these important oncogenes have been proved to promote human tumorigenesis.

Effects of endogenous sulfur dioxide on pulmonary vascular inflammation in rats with monocrotaline-induced pulmonary hypertension / 中华实用儿科临床杂志

Objective To investigate the effects of endogenous sulfur dioxide(SO2) on pulmonary vascular inflammation in rats with monocrotaline (MCT)-induced pulmonary hypertension.Methods Thirty-two Wistar rats were randomly divided into 4 groups(n =8 for each group):control group,MCT group,MCT + L-aspartic acid-β-hydroxamate(HDX) group,and MCT + SO2 group.Rats in the MCT group,MCT + HDX group,and MCT + SO2 group were subcutaneously injected with MCT(60 mg/kg) on the first day.For rats in MCT + HDX group,HDX(25 mg/kg,on day 0,7 and 14) was given orally after injection of MCT; and rats in MCT + SO2 group were subcutaneously injected with the SO2 donor sodium sulfite/sodium bisulfate(Na2SO3/NaHSO3,and mole ratio was adjusted to approximately 3:1) each day.Rats in the control group received only the same volume of solvent vehicle only.After 3 weeks,mean pulmonary artery pressure(mPAP) of each rat was evaluated by using a right cardiac catheterization procedure.Immunohistochemistry was used to detect the expression of inflammatory related factor intercellular adhesion molecule-1 (ICAM-1) and the key molecules of nuclear factor-κB (NF-κB) signal transduction pathway,including p65 and inhibitor of NF-κB (IκBα) in the small pulmonary artery endothelial cells.Results The differences in mPAP,expression of ICAM-1,IκBα and p65 in the small pulmonary artery endothelial cells were found among the 4 groups (mPAP:F =53.334,P ＜ 0.01 ; ICAM-1:F =183.82,P ＜ 0.01 ; IκBα:F =142.89,P ＜ 0.01 ; p65:F =105.46,P ＜0.01).The mean pulmonary artery pressure(mPAP) was significantly raised in MCT group rats as compared with that of the control group along with upregulated expressions of ICAM-1 protein and p65 protein in small pulmonary artery endothelial cells,while the expression of IκBα protein in small pulmonary artery endothelial cells was significantly low.After administration of HDX,the mPAP and the expression of ICAM-1 protein and p65 protein in small pulmonary artery endothelial cells further increased compared with those of MCT group,while the expression of IκBα protein in small pulmonary artery endothelial cells was significantly lower than that of MCT group.Whereas with treatment of SO2 derivatives,the mPAP,the expression of ICAM-1 protein and p65 protein in small pulmonary artery endothelial cells were significantly lower than those of MCT group,while the expression of IκBα protein in small pulmonary artery endothelial cells increased significantly compared with that of MCT group.Conclusions Endogenous SO2 might inhibit the activation of NF-κB pathway in the small pulmonary artery endothelial cells,attenuate the pulmonary vascular inflammation and prevent the MCT-induced pulmonary hypertension in rats.

The migrating pharyngeal foreign body resulted in cervical mass: one case report / 临床耳鼻咽喉头颈外科杂志

Pharyngeal foreign body is a common disease. The diagnosis and treatment are easy. However, in a few cases, pharyngeal foreign bodies migrated to other part of body, which often causing missed diagnosis or misdiagnose to delaythe treatment, and even lead to fatal complications. Here we present a case report of a 52-year-old female patient.who was found to have cervical mass 20 days before. Contrast-enhanced computed tomography showd a foreign body and foreign body granuloma on the left side of the neck. To look back on the history, the patient swallowed a fish bone in mistake one month ago.

Brain-derived neurotrophic factor transgenic neural stem cells can alleviate toxicity of β-amyloid to cultured neurons / 中华神经医学杂志

Objective To explore the resistance of neural stem cells (NSCs) in β-amyloid (Aβ) toxicity after being transfected with brain-derived neurotrophic factor (BDNF) and its mechenism.Methods NSCs in the C57 mice were transfected with BDNF gene by using lipofectamine technique; the BDNF gene expression in transfected NSCs and their differentiated cells were detected by immune-fluorescent staining and ELISA.Rat neurons were cultured in vitro and divided into 3 groups:neurons group,neurons co-cultured with NSCs group,neurons co-cultured with BDNF transfected NSCs group; two-parallel group design was adopted.Three d after the group culturing,cells from each parallel group were added Aβ40 protein.Neuron survival of each group was recorded 24 and 48 h after adding Aβ40 by ELISA.The Aβ,tau,and phosphorylating tau (ptau) levels were detected by Western blotting.Results BDNF-transfected NSCs could continuously express BDNF gene products during the first 1.5 weeks.Both transfected NSCs and their progenies expressed BDNF gene products.Using neurons group as reference,the mean survival rates of neurons plus Aβ40 group,neurons co-cultured with NSCs plus Aβ40 group,neurons co-cultured with BDNFtransfected NSCs plus Aβ40 group were 51.86％±0.03％,73.68％±0.04％ and 85.34％±0.01％ at 24 h after adding Aβ40,and 38.76 ％±0.01％,59.65 ％±0.04％,75.61％± 0.07％ at 48 h after adding Aβ40; the survival rate in the neurons plus Aβ40 group and neurons co-cultured with NSCs plus Aβ40 group was significantly lower than the other groups (P＜0.05).High level of Aβ protein was noted in the neurons plus Aβ40 group,neurons co-cultured with NSCs plus Aβ40 group and neurons co-cultured with BDNF transfected NSCs plus Aβ40 group,without significant differences between each two groups (P＞0.05).The tau protein level showed no significant differences between each two groups (P＞0.05),while the p-tau protein level was significantly different (that in the neurons co-cultured with NSCs plus Aβ40 group and neurons co-cultured with BDNF transfected NSCs plus Aβ40 group was significantly higher than that in the other groups,P＜0.05).Conclusion BDNF-transfected NSCs express BDNF gene products during the early time,which alleviate the toxicity of Aβ to NSCs and reduce the p-tau protein level,leading to increased survival rate of neurons.

A new design immunotoxin for killing high-grade glioma U87 cells: from in vitro to in vivo.

A new wave of engineered antibodies, leading to increased effectiveness of functions such as antibody-dependent cell-mediated cytotoxicity or complement-dependent cytotoxicity, is being evaluated in clinical settings. Several, such as immunotoxins, are expected to receive approval for usage soon. In this study, using a cognate heavy framework region (HFR2), two complementarity-determining regions (CDRs, i.e., LCDR1 and HCDR3) were fused to the first 388 amino acid residues of diphtheria toxin (DT388) to establish the immunotoxin IT-87. It was found that the mimetics of LCDR1-HFR2-HCDR3 retained the antigen recognition of their parent antibody. The immunotoxin IT-87 could especially kill the U87 MG glioblastoma cell line, the targets of the parent antibody, in vitro; however, the IT-87 could not kill Rajicells. In SCID mice bearing both U87 and Raji cells, the IT-87 directly targeted the U87-induced tumors (via tumor-specific surface markers) and inhibited the growth of the cells in vivo over a 20-day daily IT-87 treatment period. It is believed that the design of this particular immunotoxin could be the basis for even more promising molecules to be used in the treatment of human cancers.

Construction of a prokaryotic expression vector of human tau multi-epitope peptide and immunogenicity of the expressed product / 南方医科大学学报

<p><b>OBJECTIVE</b>To construct a prokaryotic expression vector of human tau multiepitope peptide for examining the immunogenicity of a TauP1/P2 DNA vaccine in mice using the expressed product.</p><p><b>METHODS</b>The coding sequence of Tau multiepitope peptide gene was amplified from the plasmid pVAX1-Tau by PCR and inserted into the prokaryotic expression vector pGEX-4T-2 to construct the recombinant plasmid pGEX-4T-2-TauP1/P2. The positive recombinants were transformed into E.coli BL21 cells, and the expression of fusion protein GST-TauP1/P2 was induced by IPTG and identified by SDS-PAGE. Mice was immunized with TauP1/P2 DNA vaccine and the production of the specific antibodies was detected by Dot-blot analysis using the purified fusion protein.</p><p><b>RESULTS</b>A gene fragment 300 bp in length was amplified. Enzyme digestion and DNA sequencing verified correct construction of the prokaryotic expression plasmid pGEX-4T-2-TauP1/P2. The expression of target fusion protein GST-TauP1/P2 was detected by SDS-PAGE. Specific antibodies against TauP1/P2 were detected in the serum of mice immunized with the DNA vaccine using GST-TauP1/P2 fusion protein.</p><p><b>CONCLUSION</b>The constructed prokaryotic expression plasmid of human Tau multiepitope peptide is capable of expressing the target fusion protein, which specifically recognizes the specific antibodies against TauP1/P2 in mice immunized with TauP1/P2 DNA vaccine.</p>

Motor function evaluation in rats receiving umbilical cord mesenchymal stromal cell transplantation for traumatic brain injury using CatWalk automated gait analysis system / 南方医科大学学报

<p><b>OBJECTIVE</b>To assess the effect of CatWalk automated gait analysis system for evaluation of motor function of rats with traumatic brain injury (TBI) after umbilical cord mesenchymal stromal cell (UC-MSC) treatment.</p><p><b>METHODS</b>Eighteen Wistar rats were randomized equally into normal control group, TBI ∓ saline group, and TBI ∓ UC-MSCs group. The rats in the latter two groups were subjected to weight-drop impact to induce TBI followed by injection UC-MSCs or saline into the lesion 7 days after TBI. The neurological function was assessed using CatWalk system and modified neurological severity scores (mNSS) before and 3 days after TBI and 7 days after UC-MSC transplantation. The rats were sacrificed 14 days after the cell transplantation and the brain sections were stained for immunohistochemical analyses.</p><p><b>RESULTS</b>Three days after TBI, mNSS test showed moderate injury of the rats. Seven days after the cell transplantation, the rats showed significant motor function improvement and CatWalk analysis indicated partial recovery of the gait parameters of the 4 limbs compared to the rats with saline treatment. Histological analyses showed that DiO-labeled UC-MSCs were present in the lesion boundary and expressed glial fibrillary acidic protein and β-tubulin III.</p><p><b>CONCLUSION</b>UC-MSC transplantation can promote functional improvement of the brain after TBI in rats. Compared with mNSS test, CatWalk analysis is more sensitive and objective for assessing neurological function and also provides more detailed information on specific gait parameters.</p>

Neural regrowth induced by PLGA nerve conduits and neurotrophin-3 in rats with complete spinal cord transection.

Biomaterials and neurotrophic factors represent two promising strategies for spinal cord injury repair. In this study, a combinatorial approach combining the PLGA nerve conduits and the recombinant human neurotrophin-3 (rhNT3) was utilized in a spinal cord injury animal model. After complete transection of the thoracic cord in rats, rhNT3 was administered as a single dose to the host cord caudal to a 2-mm conduit. Axonal regrowth was enhanced, as indicated by immunostaining and neurofilament-positive area measurement. Neural regrowth was further demonstrated via the retrograde tracing across the lesion. The animals implanted with the PLGA scaffold and rhNT3 exhibited significantly improved performance in BBB rating scale and grid walk tests. These observations suggest that PLGA nerve conduits combined with exogenous NT3 may serve as an alternative therapeutic approach for spinal cord injury repair.

The protective effect of EPO on Alzheimer disease rat learning and memory injury and its effect on Bcl-xl protein expression in hippocampal CA1 subregion / 中华行为医学与脑科学杂志

Objective To explore the protecting mechanism of erythropoietin (EPO) on learning and memory of Alzheimer disease (AD) rats. Methods The AD model was made by injected into rat hippocampal CA1 subregion with amyloid beta-protein(Aβ). Male Spraque-Dawley rats were as the experimental objects, which were randomly separated into 3 groups including Sham, Saline control and EPO treatment. After Aβ was injected into rats hippocampal CA1 subregion ,saline or EPO was respectively injected into the lateral ventricle of rats,with help of stereotaxic coordinates, upon the designed conditions. Hippocampal CA1 subregion Bcl-xl expression changes were observed 24 hours after the operation, and learning and memory abilities were checked 4 weeks after the operation. Results 24 hours after the operation Bcl-xl expression in the EPO group and the Saline group was less than the Sham control ,while Bcl-xl positive cells( 100.42 ± 12.43/field) in the EPO group were more than in the Saline group( 82.06 ± 19.68/field ) (P ＜ 0. 05 ). 4 weeks after the operation learning ability in the EPO group ( 20.38 ± 5.88 ) was better than Saline group ( 25.50 - 3.25 ) (P ＜ 0. 05 ), and memory ability in the EPO group (4.75 ± 1.75 ) was better than the Saline group(2.88 ± 1.55 )(P ＜ 0.05 ). Conclusion EPO could improve the learning and memory abilities in the model rats,and it could be related with EPO restraining Bcl-xl expression decreasing.

Antidepressant-like activity of tetramethylpyrazine measured by acute and chronic experimental methods in rat model of depression / 中华行为医学与脑科学杂志

Objective To study the potential antidepressive-like effect of tetramethylpyrazine (TMP). Methods Forced-swimming and chronic mild stress (CMS) tests were performed to assess the antidepressant-like activity of TMP. Male Sprague-Dawley (SD) strain rats were divided into six matched groups (n= 13 or 14 in each group) based on their sucrose consumption:control, CMS, CMS + fluoxetine, and CMS + TMP groups. The rats except control were housed separately in different rooms, and the rat model of depression was established by exposing to an unpredictable sequence of stressors for 28 days; the rats in CMS + fluoxetine were exposed to CMS and received administration of FLU (2.0 mg· kg-1·d-1 ,ig) for 28 days; the rats in CMS + TMP groups were exposed to CMS and received administration of TMP (10,20,40 mg·kg-1·d-1 ,ig) , respectively, for 28 days. The rats in control group were given ordinary daily care and received ig administration of normal saline simultaneously. The body weight, food intake and fluid consumption were measured, and the behaviors were examined by open field during the duration of the stress procedure, and forced-swimming test was performed 1 day after last unpredictable stressor. Results Acute administration of TMP markedly decreased the duration of immobility during forced-swimming test((89.0 ±37.0)s vs (117.1 ±32. 1)s, P<0.05) . Chronic administration of TMP partially countered the effects of CMS on consumption of sucrose solution and locomotion and exploration behavior, and potently shortened the immobility time during forced-swimming test following CMS in rats. The results showed that long-term administration of TMP partially reversed the effects of CMS on the body weight gain,the consumption of sucrose solution,the squares crossing in open field test and the immobility time during forced-swimming test in rats ((91.9 ±31.5) vs (124.4±27.0)s,P<0.05).Conclusion TMP shows obvious antidepressant-like activity.

Relationship of somatosensory evoked potential grading with prognosis in long-term unconscious patients after severe traumatic brain injury / 中华创伤杂志

Objective To study prognosis and grading of somatosensory evoked potential(SEP)in long-term unconscious patients after severe traumatic brain injury(TBI).Methods Five prognostic factors including age,sex,injury mechanism,history of temporal craniotomy and SEP grading were selected and analyzed in 47 patients after severe TBI with a duration of unconsciousness longer than two weeks.The prognosis was judged by Glasgow Outcome Scale.Results Prognosis was closely associated with SEP grading(P=0.024).The accuracy of SEP in assessing the prognosis was 91.5%.About 95%-100% of patients with SEP at grade Ⅲ-Ⅲ ended up with severe disability,persistent vegetative state or death.However,43.75% of patients with SEP at grade Ⅰ had good prognoses.Conclusions The SEP grading can objectively and accurately evaluate patients' prognosis and demonstrate the brain function.