Objective To clone and identify neurotrophic factor NT3 gene from mouse liver so as to establish a cell strain expressing high level of NT3 gene. Methods The target genes amplified by RT PCR were cloned into the shuttle vector pExchange 1neo, and then the DNA sequence was analyzed by enzyme digestion and sequencing. The recombinant expression vector pExchange NT3 1neo was employed to transfect the embryonic stem cell strain MESPU35, and then the transfected cells were selected by G418. The NT3 level in the transfected cells was detected by RT PCR. Results A gene fragment of 777 bp was obtained by RT PCR, and the DNA sequence was identical to mice NT 3 gene sequence of GenBank. The recombinant vector was constructed successfully and the constructed cell strain could express high level of NT 3 gene. Conclusion The successful cloning of NT3 gene from mouse liver, construction of pExchange NT3 1neo expression vector, and establishment of cell strain stably expressing high level of NT3 gene have laid a foundation for the further studies.

Objective To evaluate the inhibitory effect of butyl flufenamate (BT) on ultraviolet (UV)-induced acute skin phototoxic reaction,and to investigate its possible mechanisms.Methods Eight SKH-1 hairless mice were included in this study.The back of each SKH-1 hairless mouse was divided into six regions,which were then randomly classified into six groups:blank group receiving no treatment,UV group receiving UV radiation only,BT + UV group and vehicle + UV group topically treated with BT ointment and vehicle respectively followed by UV radiation,UV + BT group and UV + vehicle group topically treated with BT ointment and vehicle respectively after UV radiation.Skin samples were obtained from these mice at 24 hours after treatment.Subsequently,hematoxylin-eosin (HE) staining was performed,real-time PCR was carried out to detect mRNA expressions of caspase-3,p53,COX-2,PGER1,interleukin (IL)-1β,IL-6,and an immunofluorescence assay was conducted to observe the expression of caspase-3.Statistical analysis was carried out by repeated-measures analysis of variance (ANOVA).Results Compared with the UV group,both BT + UV group and UV + BT group showed a decrease in the degree of skin edema and number of apoptotic cells at 24 hours after UV radiation.Real-time PCR showed that the mRNA expressions of caspase-3,p53,COX-2,PGER1,IL-l β and IL-6 were significantly higher in the UV group than in the blank group (all P ＜ 0.05),but significantly lower in the BT + UV group than in the UV group (all P ＜ 0.05),and only the expressions of caspase-3 and p53 mRNAs were significantly decreased in the UV + BT group compared with the UV group (both P ＜ 0.05).The immunofluorescence assay revealed that the expression of caspase-3 increased in the UV group compared with the blank group,but decreased in both BT + UV group and UV + BT group compared with the UV group.Conclusion BT could partially inhibit UV-induced acute skin phototoxicity in SKH-1 hairless mice.

Objective To investigate the effects of alprostadil (Lipo-PGE) combined low-dose dopamine in the treatment of hepatorenal syndrome (HRS). Methods 61 patients with hepatorenal syndrome were randomly di-vided into two groups, treatment group 31 cases, 30 cases of the control group. Treatment group in the control group based on the 20μg soluble in 10% glucose infusion 100 ml in combined treatment with Lipo-PGE a day, one time,and combined with low-dose dopamine 3 μg/(kg·min) intravenous infusion, 1 week course of treatment. Result The treatment group liver function improvement in renal function are superior to the control group (P <0.01), to-ted effective rate was 67.7%. Conclusion The united alprostadil and low-dose dopamine satisfied with liver and kidney syndrome, there is synergy.

Objective Alprostadil can improve the clinical efficacy of the treatment of hepatitis B cirrhosis, but little literature is available about its effect on serum inflammatory factors in patients with hepatitis B cirrhosis.This study aimed to investigate the effect of alprostadil on serum inflammatory factors and liver function of the patients with hepatitis B cirrhosis and its possible action mechanisms.Methods We equally randomized 162 cases of hepatitis B cirrhosis admitted to our hospital from August 2014 to July 2015 into a control and an observation group, the former treated by conventional antiviral, liver-protecting and supportive therapies, and the latter with alprostadil in addition, both for 4 weeks.Then, we obtained the serum inflammatory factors, the contents of serum interleukin-6 (IL-6), high sensitive C reactive protein (hs-CRP) and tumor necrosis factor alpha (TNF-α) as well as such liver function indexes as glutamic-pyruvic transaminase (ALT), total bilirubin (TBil) and prothrombin activity (PTA), and compared them between the two groups before and after treatment.Results After 4 weeks of treatment, the effectiveness rate was significantly higher in the observation than in the control group (82.72% vs 62.96%, P<0.05).Compared with the baseline, the patients in the observation group showed significant improvement after treatment in serum IL-6 ([275.62±43.39] vs [97.15±19.73] pg/mL, P<0.05), hs-CRP ([425.54±55.58] vs [50.23±6.69] ng/L, P<0.05), TNF-α ([321.74±80.73] vs [85.45±13.61] pg/mL, P<0.05), ALT ([139.54±37.36] vs [96.13±23.62] μmol/L, P<0.05), TBil ([395.39±41.13] vs [271.55±25.12] μmol/L, P<0.05), and PTA ([38.50±4.19] vs [68.36±8.11]%), and the improvement was significantly better than in the control group (P<0.05).Conclusion Alprostadil helps alleviate the inflammatory condition, improve the liver function, and promote clinical efficacy in patients with hepatitis B cirrhosis.

Objective: Nowadays, the power calibration methods of the microwave hyperthermia apparatus doesn't take the power loss of the radiator into account Aiming at this problem, the authors designed an equipment of measuring the actual output power of the microwave hyperthermia apparatus. A new method is proposed for calibration the output microwave power of microwave hyperthermia apparatus. Methods: The magnetron anode current was maintained at a default value by a control system. The microwave power generated by microwave source is coupled firstly to a low-power meter by the coaxial cable to measuring the power going through coaxial cable (P_(coaxial cable)). Then the microwave radiator is connected to the coaxial cable to make the microwave radiated by radiator. The radiator is assembled in the experimental device for the microwave completely absorbed by the water. The absorbed microwave energy of the water is calculated by measuring the water temperature change. The energy loss of the experimental device is calculated using the cooling rate. The output power of the radiator is equal to the ratio of the sum of the two aforementioned energy and the time. And the efficiency of the radiator η_(radiator), is equal to P_(radiator)/P_(coaxial cable) Results: The relationship between the actual output power of the microwave hyperthermia apparatus and the mag- netron anode current is P_(radiator) = 2η_(radiator) I. The efficiency of the radiator is η_(radiator)= (34±1)%. Conclusion: From the experimental results, the current method for calibration output power of microwave hyperthermia apparatus is defective, it dose not consider the conversion efficiency of radiator. Using the calibration method introduced in this paper, wecan accurately deter- mine the actual output power of microwave hyperthermia Apparatus.

Objective To express soluble single chain variable fragments (ScFv) of monoclonal antibody MC3 recognizing colorectal carcinomas in E. coli HB2151 and to purify the soluble ScFv and identify its antigen binding activities to find new target vectors for the diagnosis and therapy of colorectal carcinomas. Methods The phage clones displaying ScFv fragment of the monoclonal antibody MC3 were used to infect E. coli HB2151 to express soluble antibodies. The soluble ScFvs were identified by Dot blot and Western blot and their antigen binding activities were determined by ELISA. The VH and VL DNAs of the ScFv DNA derived were sequenced based on the dideoxy method. Results The soluble MC3 ScFvs were expressed successfully. The expression products with a proximate MW of 32?10 3 were mainly secreted into the periplasm. The soluble ScFv containing periplasmatic extracts derived from three clones could inhibit the binding of MC3 with its antigen, and the inhibition rates were 41.19%, 36.89% and 33.77% respectively. The sequences of the VH and VL DNAs of the MC3 ScFv showed that the variable antibody genes belonged to the IgG1 subgroup and ? type. Conclusion Generation of E. coli HB2151 expressed ScFv of monoclonal antibody MC3 paves the way for further use of the antibody.

Objective To explore the efficacy of endovascular treatment for closed articular artery injuries.Methods The clinical data of 13 patients from Jan 2010 to Dec 2014 treated for closed articular artery injuries were analyzed retrospectively.The location,severity and extent of arterial injury were confirmed by intraoperative arteriography.The diameter and length of the injured arteries were measured.The guidewire was sent to the lesion site and working wire pathway was established.When the guidewire was difficult to pass through the lesion site,femoral and posterior tibial or radial artery may be used to establish working wire pathway.The stent or stent-graft was implanted through the contralateral femoral artery or distal end of ipsilateral artery.Results Intraoperative angiography confirmed intimal injury (n =2),partial transection (n =5),complete transection (n =5) and arteriovenous fistula (n =1).Eighteen stents or stent-grafts were implanted.Treatment was successful in all patients without peiropeartive death and procedure-related complications.All were followed up for 15 to 48 months and the mean follow-up was (30 ± 11) months.Three patients with stent lumen stenosis less than 50％ as showed by angiography.There was no stent fracture,displacement,or deformation.Conclusions Endovascular treatment for closed articular artery injuries is of less invasion,shorter operative time and quick postoperative recovery.

Objective To investigate the variation law of apparent diffusion coefficient (ADC) and fractional anisotropy (FA) values in patients with cerebral infarction, and to explore the relationship between the changes and the prognosis of cerebral infarct patients. Methods Sixteen patients with cerebral infarction were recruited and divided into 2 groups:good recovery and poor rehabilitation. ADC and FA values were calculated in infarct areas and control areas which were the regions with symmetrical position and the same area as infarct areas. The difference of ADC and FA values in patients at acute and earlier chronic stage between the two areas were analyzed. Results ①At acute stage, ADC values in infarct areas were lower than those in control areas (P<0.05). At early chronic stage, there was no significant difference of ADC values between infarct areas and control areas (P>0.05), moreover ADC values were higher than that at acute phase (P<0.05). ②FA values in infarct areas were lower than those in control areas at both acute and early chronic stage (P<0.05). At early chronic stage, FA values were lower than those at acute stage (P<0.05). ③There was no significant difference of ADC and FA values at both acute and early chronic stage between good recovery group compared with poor rehabilitation group (P>0.05). Conclusion There are certainly rules in changes of ADC and FA values in patients with cerebral infarction at acute and earlier chronic stage.

Objective To study on NGF positive cells changes by the action of Danshen and explore the correlation between time and amount of NGF after traumatic brain injury (TBI)in rat.Methods72 rats were randomly divided into 3 groups,including sham group,brain injury and Danshen therapy group.TBI rat models were made by fluid percussion.NGF was determined on the 3rd,7th and 14th day after TBI.ResultsNGF positive cells after traumatic brain injury in brain reached the maximum level at the 3rd day(1.29±0.61),and it decreased since the 7th day(2.03±0.42),and it decreased to the lowest level on the 14th day (0.87±0.23).NGF positive cells after traumatic brain injury treated by Danshen were more than those patients who were treated without Danshen(1.77±0.54,3.35±0.37,0.65±0.21 vs 0.75±0.31,1.58±0.52,0.49±0.36,P＜0.05～0.01).ConclusionsThe expression of NGF was significantly increased in brain damaged tissues.This may be served as a useful marker of diagnosis in time-course of brain leison.Danshen injection can be used to protect brain after traumatic brain injury.

Objective To explore the effects and mechanisms of prenatal hypoxia on vasomotor functions of fetal rats.Methods Sixteen pregnant Sprague-Dawley rats were randomly divided into two groups:control and hypoxia groups (eight in each group).Rats in the hypoxia group were provided with 10.5％ of oxygen from gestation day 5 to 21,while those in the control group were exposed to normoxic condition.Fetuses were removed from the pregnant rats by cesarean section on gestational day 21.Fetal body weight,blood gas and electrolyte levels were measured.Thoracic aorta rings were separated from fetal rats and used in different vascular function tests.Effects of hypoxia during pregnancy on angiotensin Ⅱ (Ang Ⅱ)-mediated vasoconstrictions and acetylcholine (Ach)-mediated vasodilatations in fetal thoracic aortas were measured.Changes in vasomotor functions were observed after both endothelial nitric oxide synthase (eNOS) inhibitor NG-nitro-L-arginine methyl ester (L-Name) and L-type calcium channel (LTCC) inhibitor nifedipine were administered.T-test and two-way analysis of variance were used for statistical analysis.Results (1) Compared with the control group,fetal body weight [(4.40±0.23) vs (3.33±0.42) g,t=2.871],blood partial pressure of oxygen [(50.64±2.17) vs (42.50-±-2.32) mmHg (1 mmHg=0.133 kPa),t=-2.618] and blood oxygen saturation [(58.95±1.97)％ vs (47.73±2.24)％,t=3.564] in the hypoxia group were significantly reduced (all P＜0.05).(2) Compared with the control group,Ang Ⅱ-mediated vasoconstrictions increased,but Ach-mediated vasodilatations in fetal thoracic aortas decreased in the hypoxia group (both P＜0.05).L Name induced stronger Ang Ⅱ-mediated contractions in thoracic aortas in the control group than that in the hypoxia group (P＜0.05).However,nifedipine decreased Ang Ⅱ-induced contractions,especially in the hypoxia group (P＜0.05).Conclusions Maternal hypoxia during pregnancy not only affects the growth and development of fetuses but also changes their blood vessel functions,which may be related to the change of LTCC and the impairment of eNOS.