In order to supply some reference about the construction of network course of physiology, we summed up the establishing experience from the course design, construction and application.

Objective To establish the model of serum-caused damage to pulmonary microvascular endothelial cells (PMVECs) of mice with renal ischemia-reperfusion (I/R) injury.Methods Mice PMVECs were cultured to measure the standard trans-endothelial electrical resistance (TER) in the monolayer of PMVECs.When PMVECs were cultured and arranged in compact monolayer and TER was achieved,they were divided into 4 groups (n =3 each) using a random number table:serum of normal mice group (NS group) and different concentrations (5％,10％ and 20％) of serum of mice with renal I/R injury groups (IRS5 group,IRS10group and IRS20 group).The PMVECs were cultured for 1 h in the serum-free endothelial culture medium.The 0.8 and 0.2 ml culture medium containing 20％ serum of normal mice were then added to the upper and lower chambers,respectively,in group NS.The 0.8 and 0.2 ml culture medium containing 5％,10％ and 20％ serum of mice with renal I/R injury were then added to the upper and lower chambers in IRS5,IRS10 and IRS20 groups,respectively.100 μg/ml FITC-BSA 100 μl was added to the upper chamber in the four groups.At 3,6,9,12,15,18,21 and 24 h of incubation,the PMVEC monolayer permeability (apparent permeability coefficient,Pa) was detected.Results Compared with NS group,the Pa was significantly increased at 12 and 15 h of incubation in IRS5 group,and the Pa was increased at 6-24 h of incubation in IRS10 and IRS20 groups.Compared with IRS5 group,the Pa at 21 and 24 h in IRS10 group and at 9-24 h in IRS20 group were significantly increased.Conclusion Both 10％ and 20％ serum of mice with renal I/R injury can successfully establish the model of damage to PMVECs,and 20％ serum causes a more severe damage.

Objective To investigate the efficacy of retroperitoneal laparoscopic heminephroureterectomy for duplex kidney anomalies.Methods Retroperitoneoscopic heminephroureterectomy was performed on nine patients, six males and three females.The average age of the study group was 37 years ( range 13 to 58).Seven cases had anomalies on the upper kidney pole, two cases had anomalies on the lower kidney pole.Five anomalies were on the left side, two were on the right side and two were in bilateral sides (one special case had three ureters on the left side and two ureters on the right side ).Three cases complained of flank pain; two cases were found hydronephrosis by physical routine examination;Three cases complained of flank pain and fever; one cases complained of hematuria and kidney atones.All the cases were preoperatively diagnosed by color doppler ultrasound, MRU, IVP or CTU.Retroperitoneal laparoscopic heminephroureterectomy was performed on all patients.The operation time, blood loss, hospital stay, intraoperative and postoperative complications and efficacy were observed.Results All the retroperitoneal laparoscopic procedures were successfully completed.No intraoperative complications were found.The average operation time was 87 min (range, 65 to 125).The average blood loss was 112 ml (range, 30 to 600).The recovery times of intestinal function was 1.6 days ( range, 1 to 3 ).The average postoperative hospital stay was 7 days (range, 5 to 12).The syndrome disappeared and kidney function was normal at a mean followup of 18 monthes.Conclusions Retroperitoneal laparoscopic surgeries for duplex kidney has the benefits of being minimally invasive, fewer complications, quick recovery and certainty of efficacy.Retroperitoneal laparoscopic surgeries can be considered as a first operation method to treat duplex kidney anomalies.

Objective To identify the expression of complement regulatory protein CD46, CD55, and CD59 on primary murine pallium astrocytes and the effect of inflammatory factors on it in order to lay the foundation for studying the complement system in AD. Methods The primary murine astrocytes were cultured and purified. The expression of CD46, CD55, and CD59 on the levels of mRNA and protein was assayed by immunofluorescence before and after the stimulation of LPS and IFN-?. Results The expression of CD59 mRNA was confirmed, but the expression of CD46 and CD55 was indefinite. There was no significant difference between stimulation and non-stimulation groups. Immunofluorescence results indicated that CD59 was positive, while CD46 and CD55 were weakly positive. Conclusion Protectin CD59 expresses copiously on primary murine astrocytes, which presumably protects astrocytes from the lysis of complement.

Objective To evaluate the role of Src kinase in liver injury in endotoxemic mice.Methods Forty-eight female BABL/c mice,aged 3-4 months,weighing 15-20 g,were randomly divided into 3 groups (n =16 each) using a random number table:control group (C group),endotoxemia group (lipopolysaccharide (LPS) group) and Src kinase inhibitor PP2 group (PP2 group).Endotoxemia was induced by intraperitoneal LPS 20 mg/kg in LPS and PP2 groups,while the equal volume of PBS was given in group C.In PP2 group,PP2 1 mg/kg was injected intraperitoneally at 2 h after LPS administration.At 6 h after LPS or PBS injection,8 mice in each group were chosen,and blood samples were collected from the abdominal aorta for determination of the serum levels of alkaline phosphatase (ALP).The mice were then sacrificed and livers were removed for determination of nuclear factor E2-related factor 2 (Nrf2) level,superoxide dismutase (SOD) activity,malondialdehyde (MDA) content and myeloperoxidase (MPO) activity in liver tissues.The other 8 mice in each group were sacrificed at 24 h after LPS or PBS injection,and the livers were harvested for examination of pathological changes.Results Compared with C group,the serum levels of ALP and MDA content and MPO activity in liver tissues were significantly increased,and SOD activity and Nrf2 levels in liver tissues were decreased in LPS and PP2 groups.Compared with LPS group,the serum levels of ALP and MDA content and MPO activity in liver tissues were significantly decreased,and SOD activity and Nrf2 levels in liver tissues were increased in PP2 group.The pathological changes of liver tissues were significantly attenuated in PP2 group as compared with LPS group.Conclusion Src kinase is involved in endotoxemia-induced liver injury in mice.

Objective To evaluate the effect of the serum of rats with hepato-pulmonary syndrome (HPS) on the expression of caveolin-1 and VE-cadherin in pulmonary microvascular endothelial cells (PMVECs).Methods Among the 40 healthy Sprague-Dawley rats,aged 3-4 months,weighing 220-250 g,20 rats were taken randomly for establishment the model of HPS which was produced by chronic ligation of the common bile duct,and the left 20 rats served as sham operation group.Primary PMVECs were harvested from healthy adult Sprague-Dawley rats and inoculated in ECM culture medium or on 96-well culture plate.The PMVECs of 4th-9th generation were randomly divided into 2 groups (n =36 each):control group (group C) and HPS group.In group C,the serum obtained from normal rats in sham operation group was added to PMVECs,while the serum obtained from rats with HPS was added in HPS group.The final concentration of serum was 10％.After being incubated for 12,24 and 36 h (T1-3),the expression of caveolin-1 and VE-cadherin in PMVECs was detected by Western blot,and the PMVEC adhesion rate and proliferation were determined by CKK-8 method.Results Compared with group C,the expression of caveolin-1 and VE-cadherin was significantly down-regulated,the cell adhesion rate was decreased,and the proliferation of PMVECs was enhanced in HPS group.Conclusion The serum of rats with HPS induces weakened PMVEC contact inhibition through down-regulating caveolin-1 and VE-cadherin expression.

Objective To investigate the feasibility of NR2B small interference RNA(NR2B siRNA)carried by water-soluble lipopolymer(WSLP)for treatment of neuropathic pain in rats.Methods One hundred healthy male SD rats weighing 180-200 g were randomly divided into 5 groups(n=20 each):normal control group (group C),sham operation group(group S),neuropathic pain group(group NP),group WSLP-NR2B siRNA (group W)and group WSLP-negative NR2B siRNA(group WN).Neuropathic pain was induced by partial ligation of sciatic nenre.WSLP-NR2B siRNA complex was formed by binding WSLP and NR2B siRNA.Normal saline.WSLP-NR2B siRNA complex and WSLP-negative NR2B siRNA 20μl were injected intrathecally after operation in NP,W and WN groups respectively.Mechanical withdrawal threshold(MWT)and thermal withdrawal duration (TWD)were measured before(baseline)and at 3,7,14 and 21 days after operation.Ten animals in each group were sacrificed on the 3rd day after operation and the lumbar segment(L4-6)of the dorsal root ganglia was removed for determination of the expression of NR2B mRNA and protein using RT-PCR and Western blot analysis.Results Sciatic nerve ligation significantly decreased MWT and prolonged TWD and increased NR2B mRNA and protein expression in group NP as compared with group C.WSLP-NR2B siRNA complex significantly reduced sciatic nerve ligation-induced hyperalgesia and decreased NR2B mRNA and protein expression in group W as compared with group NP.Conclusion WSLP not only mediates NR2B siRNA successfully and inhibits the expression of NR2B,but also reduces neuropathic pain in rats.

OBJECTIVE: To explore the effect of the traditional Chinese medicine Yifei Kangliu (YFKL) Oral Liquid on the proliferation, cell cycle and protein-nucleic acid synthesis of murine Lewis lung cancer cell and human lung adenocarcinoma cell line SPC-A-1. METHODS: The inhibiting rates of tumor growth were calculated by weighing the weight of tumor inoculated in vivo, combined by counting cancer cells in vitro. The ratio of the cell cycle and exponents of DNA, RNA, and protein were measured by flow cytometry (FCM). RESULTS: The inhibiting rate of tumor growth in the treated group with YFKL Oral Liquid was 30.38% (P<0.05). The proportion of cells in S phase of the treated groups with YFKL Oral Liquid was lower than that of the control group. In the group with most significant result, 72% of the cells were stagnated in G0/G1 phase. The inhibiting rates of DNA, RNA and protein in murine Lewis lung cancer were 7.4%, 23.73% and 23.31% respectively. In SPC-A-1 cell line, the inhibiting rates were 9.3%, 10.1% and 14.7% respectively, demonstrating amplified effects on lower levels. CONCLUSION: YFKL Oral Liquid significantly inhibited the proliferation of murine Lewis lung cancer cell and human lung adenocarcinoma cell line SPC-A-1 by blocking the cancer cells entering the proliferative phase resulted from its inhibition of DNA.

Using the immunohistochemical and immunoelectron microscopic methods, the localization and pattern of vasoactive intestinal polypeptidergic nerves in the wall of cerebral vessels were investigated. The results revealed that the VIP-like immunoreactive (VIPLI) fibres were located in the adventitia or at the adventitia-media border. No synaptic contacts were identified among the nerve terminals or between nerve terminal and smooth muscle cell. VIPLI terminals directly apposed to the smooth muscle cells with a distance of 100 nm.The observations mentioned above indicate that there is peptidergic innervation in the wall of the cerebral vessels besides traditional adrenergic and cholinergic innervations. The regulatory function of VIP-containing nerves to the cerebral vessels may be performed by affecting the smooth muscle directly through the nonsynaptic release. In addition, present study identified and discussed the distribution and function of substance P(SP) in the cerebral vessels.

Objective To discuss the application of spiral CT angiography in postoperative re-examination of lower limb artery stent implantation. Methods During the period from March 2012 to March 2014 at the Affiliated Nanjing Hospital of Nanjing Medical University, CT angiography was carried out in 67 patients who had received stent implantation for arteriosclerotic occlusion of lower limb. The diseases involved 78 lower limbs, and a total of 85 stents were employed. The volume rendering (VR), maximum intensity projection (MIP), multi-planar reformation (MPR) and curved surface reconstruction (CSR) were performed to stereoscopically display the lesion’s anatomy as well as the implanted stents, and the imaging manifestations were compared with the clinical symptoms and DSA findings. Results Successful examination was accomplished in 65 patients (81 stents in total), and clear images were obtained. Of the 81 stents, no stenosis was seen in 43, Ⅱ - Ⅳ grade stenosis in 32 and complete occlusion in 6. The results were closely correlated with the clinical symptoms. The CT angiography manifestations in 31 patients (34 stents in total) were compared with their DSA performed in two weeks, and the results showed that the stenotic degrees of three stents judged by CT angiography were not consistent with those judged by DSA. The consistent rate of CT angiography was 91.2% when taking DSA as the standard. Conclusion Lower limb arterial CT angiography examination is a safe and non-invasive technique, it can clearly display the stent inner canal. Therefore, this technique is of great value in postoperative re- examination of lower limb artery stent implantation.