Objective : To explore expression of related genes in breast cell line T47D induced by heregulin β1. Methods: Suppression subtractive hybridization was performed using cDNA sub traction kit to detect expression of heregulin-responsive genes in T47D cells. Results: ATP syn thase 6 was up-regulated by heregulin β1 in T47D cells at 1 and 6 hours. Conclusion: Heregulin β1 participates in the regulation of expression of both ATP synthase 6 and oxidative phosphoryla tion of T47D cells.

Objective:To investigate the efficacy of modified urethral dilatation in the treatment of female bladder neck obstruction.Methods:The clinical data of 33 female patients with bladder neck obstruction who underwent modified urethral dilatation in the Third People's Hospital of Qingdao from March 2015 to March 2020 were retrospectively analyzed. Before treatment, physical examination, routine urine examination, International Prostate Symptom Score, ultrasound examination, urodynamic examination and cystourethroscopy were performed to confirm the diagnosis. All patients were treated with α-blocker for more than 3 months, but obvious effect was not obtained. Under local anesthesia, they underwent modified urethral dilatation. After 3 months of treatment, International Prostate Symptom Score and urodynamic examination were performed to determine residual urine volume, the maximum urinary flow rate, and detrusor pressure at the maximum urinary flow rate. The curative effects of modified urethral dilatation were evaluated.Results:After modified urethral dilatation, dysuria was obviously alleviated in 25 patients. Eight patients who had no obvious improvement in dysuria were scheduled to undergo transurethral bladder neck incision. International Prostate Symptom Score after treatment was significantly lower than that before treatment [(15.18 ± 6.19) vs. (24.86 ± 7.26), t = 3.782, P < 0.001). Residual urine volume after treatment was significantly smaller than that before treatment [(53.69 ± 48.35) mL vs. (181.45 ± 92.15) mL, t = 15.328, P < 0.001]. The maximum urinary flow rate after treatment was significantly higher than that before treatment [(16.21 ± 4.22) mL/s vs. (7.91 ± 1.69) mL/s], t = 6.358, P < 0.001]. Detrusor pressure at the maximum urinary flow rate after treatment was significantly lower than that before treatment [(27.38 ± 5.13) cmH 2O vs. (57.15 ± 8.43) cmH 2O, t = 9.584, P < 0.001]. Conclusion:Modified urethral dilatation is an effective treatment method of female bladder neck obstruction. It can be used as a supplement for surgical treatment.

Objective To explore the protective effect of rhG-CSF given intranasally on cerebral infarct rats by observing the neurological dysfunction and the expression of Fas ligand (FasL) in hippocampus of cerebral infarct rats.Methods Middle cerebral artery occlusion(MCAO) model rats were established by nylon strand,reperfuse 2 hours later,and give rhG-CSF through subcutaneous and intranasal way.The rats were divided into the nermal group,the sham-operated control group(sham),MCAO group,MCAO+NS given intranasally group(NS),MCAO + rhG-CSF given subcutaneously group,and MCAO + rhG-CSF given intranasally group each group had 6 rats. At the time of 3d after reperfusion,neurological severity scores (NSS) test was performed and the expression of FasL was detected via immunohistochemical staining in collateral hippocampus. Results Neurological dysfunction appeared in all groups except for the normal and the sham group. The dysfunction of the MCAO and the NS group was the most serious,the NSS was the highest(10.20±1.85,10.30±1.76),the number of FasL positive cells was the most(41.17±3.25,41.00±2.76),and there was no obvious difference between the two groups ( P ＞0.05);the NSS and FasL positive cells decreased in the subcutaneous group(5.67±1.32,P ＜0.01;32.67±1.97,P ＜0.01) and decreased further more in the intranasal group(4.00±0.93,P ＜0.05;19.50±1.05,P ＜0.01).Conclusions rhG-CSF given intranasally can relieve the neurological dysfunction of cerebral infarct rats,and brain cells are thereby protected by resisting the expression of FasL.

Pathological pain or clinical pain is caused by tissue and nerve injuries, and is usually chronic and mainly divided into inflammatory pain and neuropathic pain. Pathological pain is typically characterized by hyperalgesia (increased responsiveness to noxious stimuli) and allodynia (painful responses to normally innocuous stimuli). The mitogen-activated proteins kinases (MAPKs) are a family of evolutionally conserved molecules that play a critical role in cell signaling, consisting of extracellular signal regulated kinase (ERK), p38, and c-Jun N-terminal kinase (JNK), which play an important role in neural plasticity of pathological pain. Inhibition of MAPKs alleviates inflammatory pain and neuropathic pain in different animal models. It is very important to study the inhibition of MAPKs as a therapeutic approach to treat pathological pain.

Objective To establish stable insect cell line expressing mIL4 on the basis of construction of inset expression vector pIZT/V5-His.Methods Amplified mIL-4 and pIZT/V5-His were treated with EcoRI and XbaI,and mIL-4 were ligated into pIZT/V5-His vector using T4 DNA ligase.Sf9 cells were transfected with recombinant DNA and ELISA was employed to detect soluble mIL-4 production by transfected Sf9 cells.Results Transfected Sf9 cells could significantly produce mIL4(1 mg?L~(-1)) compared!with control group(0.003 mg?L~(-1)).Conclusion Inset expression vector pIZT/V5-His is an ideal expression vector for mIL-4 production in transfected cells.

Objective To explore the influence of different domains of STAT6 expressed by the vector pGEX-6p-2 on signal transduction.Methods Different domains of STAT6 were amplified by PCR technique and ligated respectively into the prokaryotic expression vector.Recombinant DNA was transformed into HB101 cells.Transformants were lysed by supersonic method and lysates run on SDS-PAGE.Results Western blotting analysis showed a high level expression(60 000,56 000 and 50 000) of different domains of STAT6 SHST,DST and NST,and the empty vector only had GST expression.Conclusion Expression vector pGEX-6p-2 can be used for high level expression of different domains of STAT6.

Objective To study the expression and significance of galanin (GLA) in the prostate carcinoma (PCa).Methods The samples from 50 patients with benign prostatic hyperplasia (BPH) and 50 patients with PCa and 30 PCa patients with bone metastasis were examined by immunohistochemical staining.Results The positive rates of GLA expression in BPH,PCa,and PCa with bone metastasis were 18 ％ (9/50),68 ％ (34/50),and 80 ％ (24/30),respectively,and there were statistically significant differences between PCa patients,PCa patients with bone metastasis and BPH patients (x2 =25.5,29.74,both P ＜ 0.01),but there was no significant difference between PCa patients and PCa patients with bone metastasis (x2 =1.35,P ＞ 0.05).Conclusion GLA has higher expression in prostatic cancer cells,it might be an important indicators for differentiating prostate cancer from benign prostatic hyperplasia and predicting the prognosis of prostate carcinoma.

Objective To explore the effect of protein kinase C inhibitor on the level of phosphralated extracellular regulated protein kinases in the spinal trigeminal nucleus of migraine model rats.Methods Healthy adult male SD rats were randomly divided into four groups:normal group (group C),sham operation group (group C),migraine model group(group M),and H-7group(H-7group),with 18 rats in each group.Dural blood flow and the extracellular discharge frequency in the spinal trigeminal nucleus was recorded.ERK1/2 phosphorylation was tested.Results (1) Dural blood flow:compared with group C((3.8± 1.0)％),the dural blood flow in M group ((78.0±4.2) ％)increased obviously(P＜0.01) ; compared with M group((78.0±4.2)％),the dural blood flow in H-7 group((-24.8±4.9) ％) decreased obviously(P＜0.01).(2) The percentage of extracellular discharge frequency change:two hours after treatment,the percentage of extracellula discharge frequency change in group M ((325.9 ±47.3)％)was higher than that in group C((107.3±16.4)％).The percentage of discharge frequency change in group H-7((136.0±26.5)％) was lower than that in group M((325.9±47.3)％).There was no significant difference in the percentage of discharge frequency change between group H-7((136.0±26.5) ％) and group C((107.3 ± 16.4)％).(2) ERK1/2 phosphorylation:the ERK1/2 phosphorylation in group M was higher than that in group C.The ERK1/2 phosphorylation in group H-7 was lower than than that in group C and group M.Conclusion ERK1/2 is a downstream PKC signal path and PKC may have indirect activation of ERK1/2.

Objective To investigate the pathway of lymphatic drainage of proteins from cerebral parenchyma in subarachnoid hemorrhage rat models. Methods Healthy adult male Wistar rats were divided into Saline group, Evans blue-labeled albumin (EBA) group, and SAH + EBA group. SAH models were produced by double injection of autologous arterial blood into cisterna magna. Using a modified microinjection method, EBA was injected into left candate-putamen of the EBA group and EBA + SAH group rats. In Saline control group, saline was injected. After injection, at 12 hours, 1 day, 2 days, 3 days and 5 days, the animals were sacrificed and the fluorescence signals of EBA were imagined and analyzed along the possible lymphatic drainage pathway, e.g. the brain tissue, the wall of common carotid artery, and cervical lymphatic nodes. Results One day after injection, in EBA group, the fluorescence of EBA initially appeared on the left of the brain, the wall of common carotid artery, left lateral cerebral ventricle, and the perivascular spaces of cerebral vessels. The fluorescence signals gradually expanded to the opposite side.Large amount of fluorescence granules accumulated in the outer layer of common carotid artery. Fluorescence was also found in cervical lymphatic nodes. Two days after injection in this group, the density of fluorescencein the brain became weaker while the density of fluorescence in rhinencephalon became stronger. The fluorescence of EBA was found in lymphatic nodes adjacent to abdominal aorta. In SAH + EBA group,reduced amount and velocity of the drainage of EBA from left caudate-putamen to rhinencephalon, cervical lymphatic nodes, and lymphatic nodes adjacent to abdominal aorta were observed. From 12 hours to 5 days after injection, fluorescence intensity of EBA in deep cervical lymphatic nodes in SAH + EBA group(8.9 ±2. 0, 11.9 ± 2. 5, 17.4 ± 3.7, 26.7 ± 4. 5 and 59.0 ± 8. 1 ) were lower than those in EBA group ( 14. 5 ±3.2, 27.5 ±7.4, 60.3 ±12.3, 138.0±12.0 and 108. 1 ±13.4, F=13. 17, 24.04, 66.81, 302.77 and 59.36, P ＜ 0. 01 ). From 2 to 5 days, fluorescence intensity of EBA in lymphatic nodes adjacent to abdominal aorta was also lower in SAH + EBA group( 11.0 ± 1.5, 12. 5 ±2. 8, 23.6 ±3. 2) than those in EBA group(26. 3 ±5.9, 47.5 ±9.6, 41.0 ±9.3; F =38. 17, 72.52, 19.01, P ＜0.01). Conclusion SAH can result in reduced drainage of macromolecular substances, e.g. protein, from the brain via lymphatic pathway.

Objective To explore the effects of application calcitonin gene related peptide(CGRP) in cisterna magna on cerebral vasospasm after experimental subarachnoid hemorrhage (subarachnoid hemorrhage,SAH) in rat models.Methods 64 male Wistar rats were randomly divided into 4 groups.Group A was normal control group.After the subarachnoid hemorrhage models were established,group B,C,D were given normal saline,CGRP and adenovirus CGRP through cisterna magna respectively.White blood cells in cerebrospinal fluid were detected by automatic blood analyzer,CGRP activity was detected by enzyme linked immunosorbent assay,circulating endothelial cells were observed through laser scanning confocal microscope and parietal cortex regional cerebral blood flow were observed by laser doppler flowmeter.Basilar artery vasospasm and arterial blood gas analysis were detected by digital subtraction angiography and blood gas analyzer respectively.Results Before and after administration,there were no statistical differences in white blood cells and artery blood gas among the 4 groups (both P＞ 0.05).After administration 48 h,compared with group A,concentrations of CGRP in cerebrospinal fluid group B (0.006±0.002) did not increase (P＞0.05),but increased 200 times in Group C ((1.160±0.170) nmol/L,P＜0.05)and nearly 400 times in group D ((2.071±0.412) nmol/L,P＜0.05).Peripheral blood circulating endothelial cells count:after administration 48 h,group C((5.56±0.61) ind/0.9 μL) was less than in group B((9.94± 0.73) ind/0.9 μL).Group D((5.16±0.61) ind/0.9 μL) was less than group C(P＜0.01).Regional cerebral blood flow:after administration,compared with group B,cerebral blood flow of group C and group D increased,and the differences were both statistically significant (P＜0.01).Basilar artery diameter was detected after administration 12 h,group D ((1.000±0.025) mm) was 13％ bigger than group B ((0.670±0.028)mm,P＜0.05),3％ bigger than group C ((0.900±0.023) mm) (P＞0.05).Conclusion Cerebral vasospasm after SAH can be effectively improved by administration CGRP in cisterna magna.Adenovirus CGRP effect is better than CGRP.