Objective To elucidate the neurological m ec hanism of the referred pains in pelvic floor from prostate and the relationship between pain from prostate and pelvic floor muscle dysfunction. Meth ods The prostate was electrically stimulated,and the elicited action potentials were recorded from iliohypogastric,ilioinguinal,genitofemoral,latera l femoral cutaneous,femoral,sciatic nerves,and compound muscle action potentials (CMAP) were recorded from anal external sphincter. Results Action potentials were elicited principally in the genitofemoral nerve (93 .3%),ilioinguinal nerve (55.5%);the action potentials of the genitofemoral nerve were not influenced by transaction of the cervical spinal cord,whereas they dis appeared immediately after rat death; the action potentials were reduced conside rably after destruction of the lumbosacral sympathetic trunks.CMAPs were elicite d in the anal external sphincter in 93.3% of the rats (14/15). Conc lusions Electrical stimulation of the prostate induced reflex discha rges in the nerves to perineal,pelvic floor areas.It was postulated that the ref lex discharges are related to muscle contraction leading to referred pain in the perineal,pelvic floor areas.Pain from prostate is correlated significantly with pelvic floor muscle dysfunction.

Objective To investigate the difference of the protein expression in serum of rats with acute edematous pancreatitis (AEP) and acute necrotizing pancreatitis (ANP),and to investigate serum marker for acute pancreatitis severity.Methods The model of AEP and ANP was induced by retrograde injection of 4％ or 5％ sodium taurocholate into the biliopancreatic duct.Weak cation exchange (WCX2) and surface enhanced laser desorption/ionization time of flight mass spectrometry ( SELDI TOF MS) was used to investigate the difference of the protein expression in serum of rats with AEP and ANP.Results Thirty-eight spectral peak clusters which had significantly different signal intensities between AEP and ANP sera at mass charge ratio between 1000-50 000(P ＜0.05) were detected.The peak clusters at 9500 and 9700 in the sera of AEP were higher than that in ANP rata.Conclusions Serum analysis with SELDI-TOF MS can detect the difference of the protein expression in rats with AEP and ANP.The decreased expression of the protein of molecular weight of 9500 and 9700 may be a signal of AEP transition into ANP.

Objective To investigate the methods of prevention and treatment of presacral venous plexus bleeding in pelvic operation.Methods The clinical data of 8 cases of presacral venous plexus bleeding in pelvic operation from 1998 to 2013 were analyzed.Results All 8 cases succeeded in controlling bleeding,The amount of bleeding was 1 000-4 000 mL,the average amount is 2 600 mL.Conclusions The key to prevention of presacral venous plexus bleeding was thorough familiar with pelvic anatomy and a dexterous technique of careful dissection.Massive hemorrhage occurred direct electric coagulation hemostasis was available,the method was simple.

Objective To explore the optimal semi-quantitative uptake ratio for differentiation between benign and malignant lung lesions with 18F-FDG coincidence imaging.Methods One hundred and thirty-seven patients (89 males,48 females,age range:33-78 years) with lung diseases underwent 18 FFDG coincidence imaging.The maximum radioactivity counts of the lung lesions (T),normal chest wall soft tissues (NT1) and the contralateral lung tissue (NT2) were measured.The ratios of R1 (T/NT1) and R2(T/NT2) were calculated.The optimal threshold values of R1(cutoff)and R2(cutoff) were identified by ROC curve analysis.Using the optimal threshold,the sensitivity,specificity and accuracy were calculated.Results The optimal threshold values R1(cutoff)and R2(cutoff) were identified as 3.58 and 4.40.The sensitivity,specificity and accuracy were 90.0％(90/100),89.2％(33/37),89.8％(123/137) according to R1(cutoff) and 90.0％ (90/100),78.4％(29/37),86.9％(119/137) according to R2(cutoff) Conclusion Based on which the chest wall soft tissue is taken as a reference point,the optimal threshold value of T/NT1 is 3.58 in differentiation between benign and malignant lung lesions with 18F-FDG coincidence imaging.

Baculovirus has many advantages as vectors for gene transfer. We demonstrated that recombinant baculovirus vectors expressing p35 (Ac-CMV-p35) and eGFP (Ac-CMV-GFP) could be transduced into human kidney 293 cells efficiently. The level of transgene expression was viral dose dependent and high-level expression of the target gene could be achieved under the heterogonous promoter. MTT assay suggested that both Ac-CMV-p35 and Ac-CMV-GFP did not have cytotoxic effect on human embryo kidney 293 cells. Cell growth curve showed the Ac-CMV-p35 and Ac- CMV-GFP transduced and non-transduced cells had similar proliferation rate, so baculovirus-mediated p35expression had no adverse effect on cell proliferation. In addition, baculovirus-mediated p35 gene expression protected human embryo kidney 293 cells against apoptosis induced by various apoptosis inducers such as Actinomycin D, UV or serum-free media. These results suggested that the baculovirus vector mediated p35 gene expression was functional and it could be widely used in molecular research and even gene therapy.

Objective To construct the vector that expresses the fusion protein of HIV-Tat protein and red fluorescent protein(mCherry) in mammalian cells,and observe by fluorescence microscopy the intracellular transduction and localization of recombinant protein in cells,in order to obtain a useful tool for the study of the uptake mechanism and intracellular localization of HIV-TAT.Methods With the designed primer coding mCherry sequence,the mCherry gene was amplified by PCR with the vector pmCherry-C2 as template,and inserted into vector pET14b-His-TAT to construct the expression vector pET14b-His-TAT-mCherry.The constructed vector was then transformed into E.coli BL21(DE3),which had been identified by PCR and double digested with restriction endonuclease,followed by sequencing.After IPTG induction,the recombinant protein of His-TAT-mCherry was lyzed and analyzed with SDS-PAGE.Purified His-TAT-mCherry recombinant protein was added to Hela cells and the fluorescence was observed to evaluate the transduction efficiency.Results The results of identification by PCR,digestion with restriction endonuclease and sequencing indicated that the vector His-TAT-mCherry was correctly constructed.His-TAT-mCherry fusion protein was expressed in mammalian Hela cell line and purified successfully,and the fusion protein showed cellular transduction activity.It was found by fluorescence microscopy that the red fluorescence protein located mainly over the cytoplasm,and also the membrane to some extent.Conclusion The expression vector is successfully constructed for HIV-TAT labeled with mCherry sequence.Effective expression and purification of this fusion protein is achieved.It has been observed that the constructed vector may be expressed in mammalian Hela cell under active condition.Thus,it might be useful in the study of uptake mechanism and intracellular localization of HIV-TAT.

Objective To investigate regeneration and repair effect after ChABC,GDNF and Nogo-A Ab combination treatment for experimental spinal cord injury model.Methods Rat (T7-8 )complete spinal cord injury crosscutting animal model was established.The SD rats were randomly divided into 6 groups:normal group, sham operation group,simple transection group,A (ChABC)group,G (GDNF)group,N (Nogo-A antibody) group,and AGN (ChABC+GDNF+Nogo-A antibody)group.At 24 w after spinal cord injury,BDA tracer,NF-200,GAP-43,and GFAP immunohistochemistry were evaluated.Results BDA tracer of A group,G group and N group showed dye light,the proximal end of damaged zone showed the blue tracer particles,while damaged zone showed few blue regenerated nerve fibers.AGN group showed visible blue nerve fibers through the damaged zone and the distal segment in the damaged zone;the central zone of injury vacuolar degeneration showed the blue dyed fibers.NF-200 immunohistochemical staining showed NF-positive staining in A group,AGN was stronger than that in control group and simple transection group (P <0.05),AGN group than G group,N group significantly increased (P <0.05).GAP-43 positive staining in A group and AGN treatment group was stronger than in control group and simple transection group (P <0.05),AGN group significantly increased compared with A,G and N groups (P <0.05).GFAP positive staining in control group and simple transection group was stronger than in each treatment group (P <0.05),but A group,G group,N group and AGN group showed no significant differences (P > 0.05 ).SEP wave was detected in control group and AGN group,while the latency time was longer in AGN group than in control group.Conclusion ChABC,GDNF,and anti-Nogo-A antibody used alone or in combination can improve spinal cord injury and nerve cell function,and the joint application could improve regeneration after spinal cord injury than any monotherapy.

Objective To investigate DNA double-strand breaks and radiosensitization in renal carcinoma 786-O cells induced by fludarabine (FA) combined with different ionizing radiations.Methods The 786-O cells were exposed to FA combined with X-ray or heavy ion beam irradiation.Flow cytometry was used to evaluate the percentage of γH2AX-positive cells and cell cycle.The neutral comet assay was used to detect DNA double-strand breaks.The colony-forming assay was used to evaluate the effects of different treatments on cell survival.Comparison between groups was made by one-way analysis of variance or Dunnet' s t test.Results Compared with FA alone or irradiation alone,FA combined with different ionizing radiations increased DNA double-strand breaks as shown by significantly increased levels of γH2AX (P=0.007,0.001);FA combined with heavy ion beam irradiation lead to a cell cycle block at the radiosensitive G2/M phase and significantly increased the expression of γH2AX in the G2/M phase (P=0.000,0.000);the neutral comet assay revealed that FA combined with irradiation significantly increased DNA sublethal damage (P=0.020,0.060);FA significantly reduced the colony-forming rate after irradiation (P=0.000,0.030;0.001,0.040).Conclusions FA enhances the effects induced by X-ray and heavy ion beam irradiation with different properties.Particularly,FA substantially enhances the cell death induced by heavy ion beam irradiation.

Objective:To evaluate effect of telephone follow-up combined with written instruction on compliance and Helicobacterpylori (H.pylori) eradication in patients with H.pylori infection.Methods:A total of 160 H.pylori positive patients were randomly divided into an experimental group and a control group (n=80 in each group).Ml the patients got the guide instruction named the guidance of clinical medication for H.pylori infection patients before the treatment.The patients in the experimental group were added individualized follow-up with telephone.The compliance,eradication ofH.pylori,adverse events,and satisfaction were compared between the 2 groups.Results:The eradication rate of H.pylori in the perprotocol analysis for the experimental group and control group were 64.4％ (47/73) and 56.5％(35/62),respectively (P=0.380),while in the intention-to-treat analysis,the rates were 58.8％ (47/80) and 43.8％ (35/80,P=0.082),respectively.The compliance rate in the experimental group was significantly higher than that in the control group (91.3％ vs 77.5％,P＜0.05).There was significant difference in patients' satisfaction in good ones (75.3％ vs 51.6％) and poor ones (5.5％ vs 21.0％) between the 2 groups (P＜0.05).There were 11 patients in the experimental group and 36 patients in the control group,who appeared adverse reactions such as nausea,bad breath,abdominal distention,poor appetite,and defecation habit change during the process of eradicating H.pylori,but the occurrence rate in the experimental group was obviously lower than that in the control group (15.1％ vs 58.1％,P＜0.05).Conclusion:The telephone follow-up cannot increase the H.pylori eradication rate,but it can improve compliance and satisfaction for the patients and relieve adverse effects.

Objective To compare the effect of clustered acupuncture of the scalp combined with constraint induced movement therapy (CIMT) with that of conventional acupuncture of the body points plus conventional rehabilitation intervention after cerebral infarction using blood oxygenation level dependent functional magnetic resonance imaging (BOLD-fMRI).Methods Thirty acute cerebral infarction patients with hemiplegia were equally divided into two groups at random.The test group received the clustered acupuncture of the scalp combined with CIMT,while the control group received the body point acupuncture plus conventional rehabilitation intervention.Before and after two weeks of treatment,blood oxygenation level-dependent functional magnetic resonance imaging at 3.0 T was performed during a passive digital opposition movement.The volume and intensity of the activated areas were compared.Fugl-Meyer assessment (FMA) was carried out as well.Results The most pronounced activation appeared in the contralateral somatosensory motor cortex for both groups.There was no significant difference in average FMA scores between the two groups before the treatment,but after the treatment the averages were significantly different.An increase in activated volume and intensity in the contralateral SMC was observed in both groups after the treatment,but the test group showed a much larger change in both activated volume and intensity.Conclusion The clustered acupuncture of the scalp combined with CIMT can achieve a more significant improvement in functional recovery after acute cerebral infarction than conventional acupuncture of the body points plus conventional rehabilitation intervention.Such combined therapy is worthy of application in clinical practice.