AIM: To construct lentiviral vector carrying the angiopoietin-1(Ang-1) gene,and make it express Ang-1 in the rat mesenchymal stem cells(rMSCs).METHODS: The cDNA encoding the CDS of Ang-1 gene was obtained from the placenta of the adult Fisher 344 rats with RT-PCR.After digestion with restrication endonuclease,the Ang-1 gene was recombined to construct the transfer plasmid PNL-Ang1-IRES2-EGFP.The three-plasmid system of lentiviral vector was consisted of PNL-Ang1-IRES2-EGFP,the packaging plasmid HELPER,and the envelope plasmid VSVG,which were co-transfected to 293T cells mediated by lipofectamin2000 to produce lentiviral particles.The rMSCs were infected by obtained lentiviral particles.The insertion of Ang-1 gene was detected by PCR,the mRNA expression of Ang-1 in rMSCs was detected with RT-PCR,the protein expression of Ang-1 was observed with immunocytochemistry and Western blotting methods.RESULTS: The result of sequencing showed that the cloned Ang-1 gene was consistent with the sequence reported in GenBank.After digestion with restrication endonuclease,the 1 512 bp fragment of Ang-1 gene and the 10.5 kb vector fragment of PNL-IRES2-EGFP were observed with gel electrophoresis.The insertion of Ang-1 gene in viral genome was confirmed.The EGFP expression was observed with the fluorescent microscope.In infected rMSCs,the mRNA and protein expressions of Ang-1 were confirmed.CONCLUSION: Lentiviral vector carrying Ang-1 gene has been successfully constructed.The infected rMSCs are able to express the Ang-1 mRNA and Ang-1 abundantly.This will facilitate the following exploratory development of Ang-1 gene-modified rMSCs.

Objective:To study the effects of group counseling which based on interpersonal activities and aimed at the college students with computer gaming addiction in China.Methods:Seventeen college students with computer game addiction were chosen as the experimental group,and 10 college students from another college were chosen as the control group.The two groups had no difference in the scores of Computer Gaming Addiction Invention(CGAI)and several other scales before group counseling.There were a pre-test and a post-test separately set when the group counseling was started and finished,focused on CGAI and negative emotions and cognitions.Results:In post-test,the score of CGAI in experimental group was significantly lower than that in control group[(92.6±16.7)vs.(105.9±13.0)] and lower than that in pre-test(98.9±16.4)by analysis of covariance(Ps<0.01).The scores of Beck Depression Inventory(BDI),Self-Rating Anxiety Scale(SAS),and the negative cognition of oneself in socical settings in experimental group were also lower than those in control group [(6.4±7.9)vs.(13.5±5.6),(39.7±6.4)vs.(49.9±10.3),(15.1±4.3)vs.(19.9±3.0),Ps<0.01],while the score of Rosenberg Self-Esteem Scale(SES)was higher than that of the control group [(29.3±2.9)vs.(24.6±4.8),P<0.01].Conclusion:Group counseling based on interpersonal activities has obvious effects on the college students with computer gaming addiction.

BACKGROUND: Peripheral nerve injuries are hoped to promote the regeneration of nerve repair by elevating brain-derived neurotrophic factor (BDNF) concentration in local injury regions. OBJECTIVE: To observe the recanalization of nerve fiber and motor function in sciatic nerve injury rats following BDNF gene-modified bone marrow mesenchymal stem cells (BMSCs). DESIGN, TIME AND SETTING: The randomized, controlled animal study was performed at the Fujian Institute of Neurology from May 2007 to May 2008. MATERIALS: Femur and tibia of F344 male rats aged 2 months were sterilely harvested to prepare BMSCs. BDNF gene-modified BMSCs were prepared using constructed chronic viral vector PNL-BDNF-IRES2-EGFP. METHODS: The right sciatic nerve injury models were made using 60 adult Sprague Dawley rats. All models were randomly assigned into phosphate buffer saline (PBS) group, BMSC group and BDNF gene-modified BMSC group. 2 ?L BPS, 2 ?L BMSC solution and 2 ?L BDNF modified BMSC solution were separately transfered into injury site of each group. MAIN OUTCOME MEASURES: The retrograde horseradish peroxidase tracing was practiced to observe neural cell number in the Lumbar 4 and 5 spinal cord anterior horn at 2 and 4 weeks after microinjection. Sciatic nerve function index was used to observe rat motor function of injured limbs. Fluorescence excitation and immunohistochemistry were employed to detect BMSC survival and BDNF expression. RESULTS: Cell number was more, and nerve function recovery was better in the Lumbar 4 and 5 spinal cord anterior horn in the BDNF gene-modified BMSC group than in the PBS and BMSC groups. BMSC survival was found in the injured sites in the BMSC and BDNF gene-modified BMSC groups. BDNF expression was significantly more in the BDNF gene-modified BMSC group than in the BMSC group. CONCLUSION: BDNF gene-modified BMSCs have promotion effects on the recanalization and functional recovery of nerve fiber following peripheral nerve injury.

Objective To observe the expression level change of vascular endothelial growth factor (VEGF) in patients with obstructive sleep apnea-hypopnea syndrome (OSAHS), and to explore the relationships of VEGF expression with OSAHS, OSAHS related cardiovascular and cerebrovascular diseases. Methods Polysomnography (PSG) was used to conduct sleep apnea monitoring in 24 OSAHS patients from 6OSAHS popular families and 48 healthy controls with normal physical examination results. The expression of VEGF mRNA was examined by reverse transcriptionpolymerase chain reaction (RT-PCR) analysis, meanwhile, the level of VEGF in plasma was measured VEGF mRNA in PBMC were significantly higher in simple OSAHS group [plasma levels: (205.75±2.79) pg/ml; mRNA: 0. 61±0. 02] than in control group [(168.72±4.64) pg/ml; 0. 47±0. 02,cardiovascular and cerebrovascular diseases group [(288.74 ± 2.73) pg/ml, 1.16 ± 0. 03] than in simple OSAHS group [ ( 205.75 ± 2.79 ) pg/ml, 0. 61 ± 0.02, P ＜ 0. 01]. ( 2 ) There was a positive correlation of the levels of VEGF in plasma and mRNA with AHI as well as systolic and diastolic blood pressure in early morning. There was a negative correlation of the level of VEGF in plasma and VEGF mRNA with the lowest saturation of blood oxygen. There was a positive correlation of the level of VEGF mRNA with AHI as well as systolic and diastolic blood pressure in early morning.Conclusions The level of VEGF in OSAHS significantly increases, which may play a role in the pathophysiology of OSAHS and OSAHS related cardiovascular and cerebrovascular diseases.

Aim Toinvestigatetheeffectsoftriptonot-erpene methyl ether ( TME ) , a diterpene derived from the medicinal plant Triptergium wilfordii, on human gastric cancer AGS cell proliferation inhibition and ap-optosisinducedinvitro.Methods MTTassaywas used for screening tumor spectrum and detecting the vi-ability of AGS cells and normal human gastric epitheli-al cells GES-1 . Cell morphology was observed by light microscopy and AO / EB staining. Flow cytometry was used to detect cell apoptotic rate and cell cycle. JC-1 staining and fluorescence probe DCFH-DA were em-ployed to detect the changes of mitochondrial mem-brane potential and reactive oxygen species ( ROS ) . The effect of inhibiting AGS clonogenic survival was as-sayed by the method of plate clone formation. Western blot was used to analyse the expression of caspase-3 , caspase-8,Bcl-2andBax.Results MTTresults showed that TME exhibited significantly higher cytotox-icity to gastric cancer AGS cell line than to noncancer-ous cell line GES-1. IC50 for AGS of 48 h treatment was 23 . 85 μmol · L-1 . TME significantly inhibited colony formation and caused morphological changes in AGS cells. Annexin V-FITC / PI double staining showed the apoptotic rate increased. DCFH-DA stai-ning showed TME resulted in an increase in intracellu-lar ROS levels. Mitochondrial membrane potential de-creased after TME treatment. Western blot results showed that TME increased the proportion of Bax /Bcl-2 , with the activation of caspase-8 and caspase-3 . The broad-spectrum caspase inhibitor z-VAD-fmk pre-treatment reduced the expression of caspase-8 and caspase-3. TME enabled AGS cell cycle arrest in G0/G1phase.Conclusion TMEpossessespotenttumor selected toxicity and can induce apoptosis of AGS cells through cell cycle arrest, which is associated with Bcl-2 protein family.

To unvestugate the evolutuon law and unfluenced factors of dussolved organuc matter for electron transfer capacuty durung unutual landfull stage, dussolved organuc matter ( DOM ) was extracted from landfull wastes at dufferent depth. Shewanella oneudensus MR-1 and cutrate uron ( FeCut ) were respectuvely used as electron donor and electron acceptor to measure electron donatung capacuty, electron acceptung capacuty and electron shuttlung capacuty. Afterwards, the unfluenced factors of electron transfer capacuty were studued by analyzung spectral unformatuon. The results showed that proteun-luke components and humuc-luke components were able to transfer electrons, and they also accepted electrons from mucroorganusms. Electron donatung capacuty and electron acceptung capacuty uncreased furstly and then decreased. However, the electron shuttlung capacuty uncreased persustently durung the landfull process. Proteun-luke components were the maun components of dussolved organuc matter durung the unutual landfull stage, and ut was maunly responsuble for the electron donorung capacuty and electron acceptung capacuty of DOM. Electron shuttlung capacuty resulted from humuc-luke components durung the cycluc redox process. Electron shuttlung capacuty persustently uncreased durung the landfull process based on humuc-luke components generated durung the stage.

Pretreatment with brain-derived neurotrophic factor (BDNF) reduces ischemic damage after focal cerebral ischemia, and bone marrow mesenchymal stem cells(MSCs) were reported to ameliorate functional deficits after stroke in rats. Here we investigate the synergistically therapeutic effects of BDNF gene-modified MSCs on cerebral infarction. We transfected MSCs with the BDNF gene using a lentivirus-based system and investigated whether the BDNF-modified MSCs contributed to improved functional recovery in a rat transient middle cerebral artery occlusion (MCAO) model. Compared to untreated rats, rats that received both MSCs and BDNF-MSCs showed significantly more functional recovery. The difference in modified neurological severity score(mNSS) was statistically significant (P < 0.001). Recovery was better in BDNF-MSCs than in MSCs (P < 0.001). At the second week and second month after the systemic delivery of blank vector-modified MSCs and BDNF-modified MSCs, the treated rats exhibited more significant recovery than the control, including the accumulation and living of enhanced green fluorescence protein (EGFP)-positive cells in the infarct area and surrounding areas, neuron-like changes, expression of surface markers of neural cells, and a large amount of BDNF expression in the BDNF-MSCs-treated group. Our findings suggest that BDNF-gene-modified rMSCs can migrate to surrounding areas of the cerebral infarction lesion, differentiate into neural cells, and survive for extended periods. With the synergy of BDNF, they may promote the recovery of the neurological function following cerebral infarction and represent a new strategy for stem cell-based therapy.
Animals ; Brain-Derived Neurotrophic Factor ; biosynthesis ; genetics ; Genetic Vectors ; genetics ; Infarction, Middle Cerebral Artery ; genetics ; therapy ; Lentivirus ; genetics ; metabolism ; Male ; Mesenchymal Stem Cell Transplantation ; methods ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Rats ; Rats, Inbred F344 ; Recovery of Function ; Transfection

Objective:To investigate the effect of lappaconitine (LA) on neuropathic pain (NPP) mediated by retrograde transport of purinergic P2X3 receptor (P2X3R) in dorsal root ganglion (DRG) neurons of rats with chronic constriction injury (CCI) of the sciatic nerve.Methods:Seventy-two male healthy SD rats were selected to construct the NPP model following CCI of the sciatic nerve by ligating the right sciatic nerve. according to the random number table method. The rats were divided into CCI group, CCI+LA group and normal control group according to the random number table method, with 24 rats in each group. In normal control group, the right sciatic nerve was exposed without ligation. In CCI+LA group, the rats were given 2 g/L LA (ie, 4 mg/kg intravenously for once a day for one day only) after the same treatment as CCI group. Other two groups were injected with the identical amount of normal saline in the same way. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were evaluated before injury and at 2, 6, 12 and 24 hours after injury to evaluate the symptoms of neuralgia caused by nerve injury. The proximal and distal nerve fragments were collected in the three groups at 2, 6, 12 and 24 hours after injury. Western blotting was applied to analyze the expression of P2X3R at 2, 6, 12 and 24 hours after injury and the expression of neurotrophic factor (NGF) and tyrosine kinase receptor A (TrkA) at 24 hours after injury to evaluate the effect of LA on P2X3R, NGF and TrkA.Results:There were insignificant differences in MWT and TWL among all groups before injury (all P>0.05). Compared with normal control group, MWT and TWL were significantly decreased in CCI group and CCI+LA group at 2, 6, 12 and 24 hours after injury (all P<0.05 or 0.01). There were insignificant differences in MWT and TWL between CCI group and CCI+LA group at 2 and 6 hours after injury (all P>0.05), while MWT and TWL were significantly higher in CCI+LA group than those in CCI group at 12 and 24 hours after injury (all P<0.05 or 0.01). In the proximal sciatic nerve segment, Western blotting showed similar levels of P2X3R among all groups at 2, 6, 12 and 24 hours after injury (all P>0.05). In the distal sciatic nerve segment, Western blotting showed higher expression of P2X3R in CCI group than that in normal control group at 2, 6, 12, 24 hours after injury (all P<0.01), higher expression of P2X3R in CCI+LA group than that in normal control group at 2, 6 and 12 hours after injury (all P<0.05), similar expression of P2X3R expression between CCI+LA group and normal control group at 24 hours after injury ( P>0.05), similar expression of P2X3R between CCI group and CCI+LA group at 2 and 6 hours after injury (all P>0.05), and lower expression of P2X3R in CCI+LA group than that in CCI group at 12 and 24 hours after injury ( P<0.05 or 0.01). In the proximal and distal nerve fragments, the expression of NGF was lower in normal control group than that in CCI group and CCI+LA group ( P<0.05 or 0.01), but was similar between CCI group and CCI+LA group at 24 hours after injury ( P>0.05). In the proximal and distal nerve fragments, there were insignificant differences in the expression of TrkA among all groups at 24 hours after injury (all P>0.05). Conclusion:Early LA treatment after injury can alleviate mechanical and thermal hyperalgesia in NPP rats, which may be related to the reduction of P2X3R retrograde transport in DRG neuron axonal.

Objective:To explore the effect of perioperative whole-course closed-loop temperature management system for prevention of hypothermia in elderly patients with femoral neck fracture.Methods:A retrospective case-control study was conducted to analyze the clinical data of 120 elderly patients with femoral neck fracture admitted to Chengdu First People's Hospital from January 2020 to December 2020. There were 48 males and 72 females, aged 60-79 years［（68.3±5.0）years］. All patients underwent total hip arthroplasty. Heat warming with the whole-course closed-loop temperature management system was applied in 40 patients（closed-loop group）, with the medical heating mattress in 40 patients（heat preservation group）, and with the quilt in 40 patients（control group）. The core body temperature of the three groups was recorded 8 minutes after wearing the temperature sensor（T0）, immediately after induction of anesthesia（T1）, 30 minutes after induction of anesthesia（T2）, 60 minutes after induction of anesthesia（T3）, at the end of surgery（T4）, and 20 minutes after entering the postanesthesia care unit（PACU）（T5）. The thrombin time（TT）, prothrombin time（PT）and activated partial thromboplastin time（APTT）were compared among groups before operation and at T4. The incidence of intraoperative hypothermia, incidence of shivering during PACU, incidence of surgical site infection, extubation time（from the day of drug withdrawal to the time of tracheal tube removal）, length of hospital stay（from the day of operation to the time of discharge）and adverse reactions related to the devices were compared among groups.Results:There was no statistically significant difference in core body temperature among three groups at T0（ P>0.05）. The core body temperature in closed-loop group［（36.61±0.28）℃］was higher than that in heat preservation group［（36.45±0.28）℃］and control group［（36.44±0.27）℃］at T1（ P<0.05）, but no statistically significant difference was found in core body temperature between heat preservation group and control group（ P>0.05）. The core body temperature in closed-loop group［（36.46±0.28）℃,（36.40±0.30）℃,（36.45±0.37）℃,（36.50±0.27）℃］was higher than that in heat preservation group［（36.32±0.31）℃,（36.24±0.26）℃,（36.28±0.30）℃,（36.24±0.31）℃］and control group［（36.12±0.30）℃,（35.98±0.28）℃,（35.73±0.31）℃,（35.81±0.32）℃］at T2-T5（ P<0.05）, and a higher temperature was found in heat preservation group than control group（ P<0.05）. The core body temperature in closed-loop group was increased at T1 compared to that at T0（ P<0.05）, while there was no statistical difference at T0 and T1 in heat preservation group and control group（ P>0.05）. The core body temperature in closed-loop group showed no significant difference at T2-T5 and T0（ P>0.05）, while the core temperature in heat preservation group and control group was decreased at T2-T5 compared to that at T0（ P<0.05）. The core body temperature in closed-loop group was within the set target temperature range. There was no statistically significant difference in TT, PT and APTT among the three groups before operation（ P>0.05）. At T4, the levels of TT, PT and APTT in closed-loop group［（18.9±1.7）seconds,（11.9±0.7）seconds,（35.5±3.3）seconds］and heat preservation group［（18.8±1.7）seconds,（11.6±0.8）seconds,（32.8±2.7）seconds］were shorter than those in control group［（20.9±1.3）seconds,（13.0±0.8）seconds,（35.7±3.4）seconds］（ P<0.05）. At T4, the levels of APTT in closed-loop group and heat preservation group were longer than those before operation（ P<0.05）. At T4, the levels of TT, PT and APTT in control group were longer than those before operation（ P<0.05）. There was no significant difference in the incidence of intraoperative hypothermia or shivering during PACU between closed-loop group［18%（7/40）, 3%（1/40）］and heat preservation group［33%（13/40）, 8%（3/40）］（ P>0.05）, which were lower than those in control group［75%（30/40）, 33%（13/40）］（ P<0.05）. There was no significant difference in the extubation time between closed-loop group［（12.5±3.6）minutes］and heat preservation group［（13.2±3.9）minutes］（ P>0.05）, which was shorter than that in control group［（16.6±4.0）minutes］（ P<0.05）. The incidence of surgical site infection and length of hospital stay were statistically insignificant among the three groups（ P>0.05）. No adverse reactions such as allergic reaction or pressure ulcers occurred in the three groups. Conclusion:For the elderly patients with femoral neck fracture, the perioperative whole-course closed-loop temperature management system can effectively maintain the core body temperature, improve coagulation function, prevent hypothermia, reduce shivering and shorten the extubation time, without increasing the risk of other adverse events, so the system can be safely used in clinical practice.

Objective To observe the effect of massage at acupoints on head, neck and face on dysphagia after stroke. Methods 60 stroke patients with dysphagia were randomly divided into treatment group (n=30) and control group (n=30). The control group received conventional rehabilitation, including acupuncture and swallowing training, and the treatment group received acupoint massage on head,neck and face in addition. They were assessed with Standardized Swallowing Assessment (SSA) before and 4 weeks after treatment. Results The score of SSA improved in both groups (P<0.001), and improved more in the treatment group than in the control group (P<0.001) after treatment. Conclusion Conventional rehabilitation with acupoint massage is more effective on dysphagia after stroke.