Objective To investigate the practical efficacy and safety of ultrasound with microbubbles mediated rAAV2-EGFP to retina of rat after intravitreal and subretinal injection. Methods Gene transfer was examined by rAAV2-EGFP intravitreal and subretinal injection into the Wistar rats with or without microbubbles. The eyes were exposed to US (1 MHz,2 W/cm2, duration 5 minutes,duty cycle 50%,pulse recurrent frequency 100 Hz). The onset of EGFP gene expression, lightness of fluorescence, area of fluorescence and its distribution in the fundus in vivo via fluorescence stereosocope were investigated on the 4th,7th, 35th,49th and 120th day respectively. The value of gene transfer was quantified through the EGFP fluorescence quantitative methods by Axiovision 3. 1 software. HE staining was used to observe tissue damage. Results There was no fluorescence observed by fluorescence stereosocope after intravitreal injection after two-month study. After subretinal injection, ultrasound-targeted microbubbles destruction (UTMD) strongly increased gene transfer efficiency. UTMD used in the experiment did no harm to the rat retina structure. Conclusions UTMD could not enhance rAAV2-EGFP transfecion efficiency to rat retina after intravitreal injection but the transduction could be enhanced significantly after subretinal injection.

Objective To evaluate the effect of dexmedetomidine on synaptic transmission in the spinal dorsal horn of rats.Methods Male Sprague-Dawley rats,aged 4-6 weeks,weighing 150-200 g,were used in the study.The lumbar enlargemnent segments of the spinal cord were harvested,and the parasagittal lumbosacral spinal cord slices with attached dorsal roots were prepared and incubated in artificial cerebro-spinal fluid.The whole-cell patch-clamp technique was used to record each index,and 4 spinal cord slices were selected and used for each index records.Experiment Ⅰ Dexmedetomidine was added cumulatively in concentration increments.Aδ and C fibers-mediated evoked excitatory postsynaptic currents (eEPSCs) were recorded before administration (baseline) and during perfusion with dexmedetomidine 4 and 10 μg/ml.Experiment Ⅱ The neurons innervated by Aδ and C fibers were selected,and Aδ and C fibers-mediated eEPSCs were recorded before administration (baseline),at 5 min of perfusion with yohimbine (alpha 2 adrenergic receptor antagonist) 2 μmol/L,and during continuous perfusion with yohimbine 2 μmol/L plus dexmedetomidine 4 μg/ml.Experiment Ⅲ The evoked excitatory postsynaptic potentials (eE-PSPs) and evoked inhibitory postsynaptic potentials (eIPSPs) were recorded before administration (baseline) and during perfusion with dexmedetomidine 4 μg/ml.Results Dexmedetomidine could dose-dependently inhibit Aδ and C fibers-mediated eEPSCs,dexmedetomidine could inhibit Aδ and C fibers-mediated eEPSPs and produced no effect on eIPSPs,and yohimbine could inhibit dexmedetomidine-induced inhibitory effect on eEPSCs.Conclusion The mechanism by which dexmedetomidine inhibits nociceptive information transmission in the spinal dorsal horn is related to inhibition of excitatory synaptic transmission through activating α2-adrenergic receptors,but not related to activation of inhibitory synaptic transmission in rats.

Objective To explore the intervention mechanism of Yiqi Huoxue Fang on deep venous thrombosis ( DVT) of rabbit model,by observing its effects on serum t-PA and PAI-1,D-D,NO,CRP,PT,blood rheology and vascular pathology. Methods Fifty New Zealand rabbits were randomly selected.Ten rabbits were served as blank control group.DVT animal models were established in the remaining 40 rabbits, which were randomly divided into model control group, Yiqi Fang group, Huoxue Fang group and Yiqi Huoxue Fang group (n=10 each group).The changes of serum t-PA,PAI-1,D-D,NO,CRP,PT and pathological changes of blood vessel were observed after 7-days treatment. Results As compared with the blank control group, the serum t-PA,PAI-1,D-D,CRP and PT were significantly increased in the model control group,Yiqi Fang group,Huoxue Fang group and Yiqi Huoxue Fang group (all P<0.01); NO was decreased (P<0.01 or P<0.05 in model control group,Yiqi Fang group and Huoxue Fang group;P>0.05 in Yiqi Huoxue Fang group vs.the blank control group).As compared with model control group,the serum t-PA,NO,D-D increased,PAI-1 and CRP value reduced in Yiqi Huoxue Fang group and Huoxue fang group (P<0.01 or P<0.05).As compared with Yiqi Huoxue Fang group,serum t-PA in both Yiqi Fang group and Huoxue Fang group was significantly lower (P<0.01 and P<0.05) ,other indexes were significantly higher than that in Yiqi Huoxue Fang group (serum PAI-1,P<0.01 in Yiqi Fang group;NO and D-D,P<0.01 or P<0.05 in Yiqi Fang group;CRP value,P<0.01).Vascular pathology showed that blood vessels,especially vascular endothelia,of Yiqi Huoxue Fang group were most slightly injured,with slightest infiltration of inflammatory cells. Conclusion Yiqi Huoxue Fang can prevent DVT through regulating t-PA and PAI-1,keeping the fibrinolytic system function at a relatively high level,protecting vascular intima and relieving the inflammation in venous thrombosis.

Objective: α-asarone is a major effective component that can be isolated from Acorus tatarinowii Schott,a Chinese herbal medicine. Clinical investigations have shown that α-asarone has strong sedative and anti-convulsive action in the central nervous system. In recent years, several medicines containing a-asarone were applied in treatment of asthma, bronchitis, expectorant, or epilepsy. However, the underlying cellular mechanism of ct-asarone is still unknown. Here the authors considered EAAC1, the transporter for the excitatory glutamate, as a possible target. Methods: Supercritical CO2 fluid extraction and silica gel column chromatography were used to obtain ct-asarone from the rhizomes of Acorus tatarinowii Schott. Xenopus oocytes with heterologously expressed EAAC 1 were used as a model system. Rate of glutamate uptake was measured by means of isotopic tracer technique. Glutamate-induced current was recorded under two-electrode voltage clamp. 40μg/mL of ct-asarone was used for testing its effect on EAAC1 activity. Results: ct-asarone induced a slight, but still significant stimulation of rate of glutamate uptake by 15%. In contrast, EAACl-mediated current became reduced (by 30% at -100 mV). Since EAAC 1 can operate in transport and also in an ion-channel mode, the result indicates strong inhibition of the channel mode. This inhibition is voltage-dependent becoming larger at more negative potentials. Conclusion: The stimulation of glutamate uptake reduces glutamate concentration in the synaptic cleft and, hence, reduces excitatory synaptic activity. The inhibition on the ion-channel mode stabilizes the membrane potential, and therefore, also contributes to reduced excitatory activity.

Objective:The standard treatment for inoperable locally advanced esophageal cancer is concurrent chemoradiotherapy, but the survival was not satisfied. Nituzumab is a humanized IgG monoclonal antibody against EGFR. The purpose of this study is to investigate the toxicity and efficacy of concurrent chemoradiotherapy combined with nituzumab for locally advanced esophageal cancer.Methods:We retrospectively reviewed the clinical data of locally advanced esophageal cancer who were treated with concurrent chemoradiotherapy combined with nituzumab in Peking University Cancer Hospital from June 2015 to June 2020. Kaplan- Meier method was used for analysis. Results:Thirty Patients were enrolled this study.After a median follow-up of 22.5 months, The objective response rate was 93%. The 1-year, 2-year, 3-year overall survival rates were 83%, 57% and 41%, with the progression-free survival rates 75%, 47% and 32%, with the local-recurrence free survival rates 83%, 53% and 37%, with the metastasis-free survival rates 75%, 51% and 36%, respectively.The incidence of grade≥3 hematological toxicity was 32%. There were 16% patients experiencing grade≥3 esophagitis.Conclusion:The preliminary result of concurrent chemoradiotherapy combined with nituzumab is effective and safe for patients with locally advanced esophageal cancer.

Objective To evaluate whether S-100β protein could be a serum marker for traumatic spinal cord injury (SCI). Methods From June, 2013 to October, 2014, 24 patients with complete SCI were measured the serum S-100β protein concentrations with en-zyme-linked immunosorbent assay, one week, three and six months after SCI. Serum from ten healthy persons was as normal control. Re-sults The serum S-100βprotein concentrations increased one week and 3 months after SCI (Z>4.273, P<0.001). Conclusion The increase of serum S-100βprotein may help assessing early impairment after complete SCI.

Objective: To investigate the possible mechanisms in acupuncture analgesia by interaction of &opioid receptor with neurotransmitter transport proteins or the Na+-K+pump. Methods: Microinjection of respective heterologous cRNA into the Xenopus oocytes as a model system, and measurement of steady-state currents under two-electrode voltage clamp. Results: The co-expression of the δ-opioid receptor with GAT1, EAAC1 or the sodium pump resulted in reducing activity of the respective transporter. Opioid receptor activation affected transporter activity in different ways: 1) GAT1 was further inhibited; 2) EAAC1 was stimulated; 3) Na+-K+ pump activity interfered with agonist sensitivity of DOR. Pump inhibition led to higher sensitivity for DPDPE. Conclusion: GABA transporter inhibition and glutamate transporter stimulation may counteract pain sensation by affecting the neurotransmitter concentration in the synaptic cleft and, therefore, may contribute synergistically to pain suppression by acupuncture. Sodium pump inhibition by endogenous ouabain may amplify these effects. These synergistic effects may be the molecular mechanism of inhibiting pain sense and/or acupuncture analgesia.

Objective:To assess the validity and reliability of the Chinese version of the eleven-item Kutcher Adolescent Depression Scale (KADS-11)in Chinese adolescents,calculate its optimal cut-off value and the sensi-tivity and specificity,and explore the possibility of providing a useful tool to assess the severity of adolescent de-pressive symptoms.Methods:Totally 3180 students aged 11 -17 years were selected from schools in 6 provinces and Shanghai.All of them were asked to complete the KADS-11 and Children Depression Inventory (CDI). Students whose CDI scores were above 19 (including 19)were diagnosed with the DSM-IV criteria of depressive disorder,73 students from Shanghai sample were assessed with KADS-11 and CDI to analyze the test-retest reliabil-ity 1 month later.Results:Exploratory factor analysis showed that KADS-11 had 2 factors,and confirmatory factor analysis tested proved the 2-factor model fit better than the one-factor model.The KADS-11 total scores were posi-tively correlated with CDI total scores (r =0.74,P <0.01 ),and the KADS-11 scores were higher in depressive group than those in non-depressive group.The mean area under the curve (AUC)of KADS-11was 0.94,the mean area under the curve of each item ranged from 0.7 to 0.9.The optimal cut-off point of KADS-11 was total score≥9,sensitivity and specificity were 89% and 90% respectively.The Cronbach's alpha coefficient of the KADS-11 was 0.84,the spilt-half reliability coefficient was 0.71 (P <0.01),and the test-retest coefficient was 0.77 (P <0.01).Conclusion:The KADS-11 is appropriate for Chinese adolescents because of its good validity,reliability and diagnosis accuracy,it could be used to assess depressive symptoms for adolescents.

Cystic fibrosis (CF) is an autosomal recessive inherited disease affecting multiple body systems.Pseudo-Bartter syndrome (PBS) is a common manifestation of CF, with such clinical features as hypochloremia, hyponatremia, hypokalemia and metabolic alkalosis.However, PBS patients do not have renal tubulopathy.Children with CF are prone to develop electrolyte abnormalities due to fluid and electrolyte loss.In this article, the pathogenesis, clinical manifestations, diagnosis, and treatment of CF associated PBS were reviewed in order to enhance clinical understan-ding of this disease.

Objective The primary aim of this study was to examine the proportion and natural history of Helicobacter pylori (Hp) negative bleeding peptic ulcers. Methods The study was designed as a multiple-center, case-control study conducted in 14 endoscopy centers in China from April 2006 to March 2007. Each center was expected to recruit 30 peptic ulcer patients with bleeding ( PUB group) and 30 without (PU group). All screened patients with upper gastrointestinal bleeding received endoscopy within 24 hours of admission. Biopsy specimens were taken from the antrum to determine Hp infection by rapid urease test and pathology. Patients with negative Hp infection at first examination were asked to receive urease breathe test (UBT) one month later. Results A total of 617 patients were enrolled with 263 in PUB group and 354 in PU group. There is no significant difference in demographic characters between 2 groups ( P ＞0. 05). The rate of Hp infection in PUB group ( 161/263, 61.2% ) was significantly lower than that in PUgroup (311/354, 87. 9%, P ＜0. 001 ). The incidence of complex ulcer in Hp positive PUB patients was 7.5% ( 12/161 ), which is significantly higher than that in Hp negative PUB patients ( 1/102, 1.0% , P =0. 018). In PUB group, no significant differences were found between Hp positive and negative patients in regarding of age, sex, rates of haematemesis, duodenal ulcer and gastric ulcer, and size of ulcer ( P ＞0. 05 ). Among 102 Hp negative cases in PUB, no positive case was found in UBT one month later. Conclusion We have demonstrated a rise in the incidence of Hp negative bleeding ulcers in China. The idiopathic ulcer was not rare, and might have a higher tendency to cause bleed.