Adult ; Female ; Hemoptysis ; etiology ; Humans ; Pulmonary Artery ; abnormalities ; Recurrence ; Tomography, X-Ray Computed

Objective To observe the effect of Ginkgo biloba extract (EGb) on intestinal function after spinal cord injury (SCI) in rats. Methods 36 Sprague-Dawley rats were randomly divided into group A (n=12), group B (n=12) and group C (n=12). SCI model was established with Allen's mode (10 g×25 mm) at T10. 30 minutes later, group A was intraperitoneally injected with methylprednisolone 30 mg/kg every 24 hours; group B was injected with Shuxuening injection (EGb) 1.75 mg/kg every 24 hours; group C were injected with equal volume of saline. The slow wave of intestinal smooth muscle was measured, the activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) in serum were determined 1 day, 3 days and 7 days after modeling, while intestinal tissue was tested with HE staining. Results The amplitude and frequency of the myoelectric slow wave increased in the groups A and B 3 and 7 days after modeling compared with those in the group C (P<0.05); meanwhile, the activity of SOD increased and content of MDA decreased in the groups A and B (P<0.05). The HE scores decreased in the groups A and B compared with those in the group C (P<0.05), which presented that the inflammatory exudation was mild, the hemorrhagic spot was few and the area was limited. The intestinal villous of the group C was blunt with large infiltration of inflammatory cells and inflammatory exudate on the mucosal surface. Conclusion EGb can improve the recovery of intestinal function in rats spinal cord injury through antioxidant.

Objective To explore the mechanism and effect of miR-146a on alveolar macrophages and to observe the changes of miR-146a expression in the LPS-induced alveolar macrophages. Method NR8383 alveolar macrophages were divided into LPS-stimulated group and control group, and the cells of former group were treated with LPS ( 1 μg/mL) and then incubated for 3 h, 6 h and 12 h, respectively. The level of TNF-α in the supernatant of cells was assayed by using enzyme-linked immunosorbent assay (ELISA), and the expression of miR-146a of cells was detected by using Real-Time PCR (TaqMan probe).Statistical analysis carried out by using SPSS 13.0 software package in which One-way ANOVA and Student's t-test were used. Results Compared with control group, the levels of TNF-α in the supernatant of cells were significantly increased 3 h, 6 h and 12 h after LPS challenge (P ＜ 0.01 ). The expression of miR-146a increased 6 h and 12 h after LPS stimulation in NR8383 cells( P ＜0.01 ), and it had an upward tendency.Conclusions The expression of miR-146a in alveolar macrophages increases after LPS-stimulation. It hints miR-146a may be involved in the regulation of the inflammatory responses produced by alveolar macrophages.

Objective To explore the regulation mechanism of X box binding protein 1 (XBP1)signal transduction pathway for TNF-α and effective approach in ischemia/reperfusion (I/R) injury of liver transplantation for short hairpin RNA (shRNA) interference used to gene therapy in liver graft.Methods Male Sprague-Dawley rats were divided into three groups: the cold ischemia transfection group (CIT), the in vivo transfection group (IVT) and the control group. Experiments of orthotopic liver transplantation were performed by two cuff method. The rats in CIT were perfused with XBP1-shRNA plasmid (pSIXBP1) during cold ischemia phase, those in IVT received the equivalent volume (2 ml) of pSIIRAK 4 after portal vein inoculation, and those in the control group were not subjected to any treatment. Rats were killed at 60 or 180 min after restoring reperfusion of hepatic portal vein.Histopathological damage degree of graft liver was observed by light microscope. The expression levels of XBP1 gene and protein were detected by RT-PCR and Western blotting. The activities of NF-κB and the serum TNF-α level were detected by ELISA. Results All the indexes including the degree of histopathological damage, the expression levels of XBP1 mRNA and protein and the TNF-α level were significantly decreased in CIT as compared with IVT and control group (P＜0. 05). However,there was no significant difference in NF-κB activity among the three groups (P＞0. 05). Conclusion The role of XBP1 pathway in TNF-α gene regulation and that of NF-κB pathway in rat liver I/R injury are two relatively independent aspects, and the depression of XBP1 expression with XBP1 shRNA through portal vein perfusion during cold ischemia phase could effectively alleviate graft hepatic I/R

Objective:To investigate the effect and mechanism of exosomes derived from human-induced pluripotent mesenchymal stem cells (iMSC-Exos) on alveolar macrophages (AM) pyroptosis.Methods:The exosomes in the culture supernatant of human-induced pluripotent mesenchymal stem cells (iMSC) were extracted by rotating ultrafiltration, and the extracted exosomes were identified by transmission electron microscopy, Western blotting and high-resolution adjustable resistance pulse. The rat alveolar macrophage cells (NR8383 cells) were cultured in vitro and the logarithmic growth phase cells were divided into three groups: the control group was added with an equal volume of phosphate buffered saline (PBS) in the AM supernatant; in LPS/ATP group AM cells were stimulated with 500 μg/L LPS for 23 hours and then 5 mmol/L ATP was added for 1 hour to induce pyrolysis; iMSC-Exos group was incubated with AM and 100 mg/L iMSC-Exos for 3 hours before giving LPS and ATP. The cytotoxic activity was detected by cell counting kit-8 (CCK-8) and lactate dehydrogenase (LDH) analysis, the apoptosis and the expression of caspase-1 were observed by immunofluorescence, the levels of inflammatory factors interleukins (IL-1β and IL-18) released by AM were detected by enzyme linked immunosorbent assay (ELISA), the NOD-like receptor protein 3 (NLRP3) inflammasome pathway and the expression level of pyroptosis related protein gasdermin D (GSDMD) were detected by Western blotting. Results:The extracted exosomes were observed by transmission electron microscopy as round vesicles, expressing exosomal markers CD63 and CD9 showed by Western blotting, high-resolution adjustable resistance pulse showed the average diameter of the particles was 130 nm, and could be uptaken by AM. Compared with the control group, the cell activity decreased [(0.56±0.05)% vs. (1.06±0.07)%, P < 0.01], the release of necrotic substance LDH increased (U/L: 1 218.86±22.73 vs. 188.30±1.61, P < 0.01), the expression levels of inflammatory factors increased [IL-1β (ng/L): 958.91±32.78 vs. 194.63±5.14, IL-18 (ng/L): 870.89±21.86 vs. 288.85±24.48, both P < 0.01], and the apoptosis rate [(55.35±6.19)% vs. (12.01±1.32)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 41.06±3.65 vs. 2.80±0.54, P < 0.01) elevated in the AM after LPS/ATP stimulation, suggesting that LPS combined with ATP successfully induced alveolar pyroptosis. Compared with the LPS/ATP group, AM pretreated with iMSC-Exos showed increased cell viability [(0.81±0.05)% vs. (0.56±0.05)%, P < 0.01], decreased LDH secretion (U/L: 535.05±42.55 vs. 1 218.86±22.73, P < 0.01), decreased expression of inflammatory factors [IL-1β (ng/L): 381.82±19.50 vs. 958.91±32.78, IL-18 (ng/L): 533.77±31.54 vs. 870.89±21.86, both P < 0.01], and decreased apoptosis rate [(19.74±2.96)% vs. (55.35±6.19)%, P < 0.01] and caspase-1 expression (fluorescence intensity: 12.16±1.31 vs. 41.06±3.65, P < 0.01). At the same time, the expression of NLRP3 inflammasome pathway [NLRP3 protein (NLRP3/β-actin): 0.62±0.06 vs. 1.89±0.11; cleaved caspase-1 protein (cleaved caspase-1/β-actin): 0.42±0.07 vs. 1.22±0.17, both P < 0.01] and pyrolysis-related protein was significantly inhibited [GSDMD protein (GSDMD/β-actin): 0.57±0.05 vs. 1.22±0.05, P < 0.01]. Conclusion:iMSC-Exos successfully reversed the AM pyroptosis and inflammatory factor expression induced by LPS/ATP, which may be due to the targeted inhibition of NLRP3 inflammasome pathway, suggesting that iMSC-Exos can exert anti-inflammatory effects by inhibiting the pyrolysis of AM.

Objective To construct recombinant Mycobacterium smegmatis vaccine expressing Cysticercus cellulosae cC1 anti-gen. Methods The recombinant pET28a-cC1 plasmid was extracted and double digested by Xho I and BamH I restriction en-zymes,and shuttle plasmid pMV261 was extracted and double digested by Hind III and BamH I restriction enzymes. Both frag-ments were modified by Klenow fragment to form blunt end,then the large fragments of cC1 and pMV261 plasmid were purified and ligated by T4 ligase enzyme. The recombinant pMV261-cC1 plasmid was constructed and sequenced. Then the pMV261-cC1 plasmid was transformed into Mycobacterium smegmatis by the electrotransformation method. The recombinant cC1-Mycobacterium smegmatis was induced by heat and identified by the Western blotting method with the sera of cysticercosis patients. In addition, the growth states of the Mycobacterium smegmatis and the recombinant cC1-Mycobacterium smegmatis were compared and the growth curves were drawn. Results The restriction enzyme and sequencing results showed that the recombinant pMV261-cC1 plasmid was successfully constructed. After heat induction,a 40 kD band was showed by PAGE analysis of cC1-Mycobacterium smegmatis. The Western blotting results showed that the sera of cysticercosis patients could recognize the 40 kDa band,which sug-gested that cC1 protein was expressed in Mycobacterium smegmatis. Compared with the Mycobacterium smegmatis,the recombi-nant cC1-Mycobacterium smegmatis showed no significant difference in proliferation characteristics. Conclusion The recombi-nant cC1-Mycobacterium smegmatis vaccine has been successfully constructed.

OBJECTIVETo assess the functional result of a "Two in One" urethroplasty which combined oral mucosa graft and local flap.

METHODS17 patients with hypospadias underwent a "Two in One" urethroplasty, which combined buccal mucosa and local flap for urethral reconstruction. Uroflowmetry was performed 1 day before and 1 year after operation. The urine flow rate, voided volume and urine flow curves were detected using a rotating sensor. The results of maximum urine flow rate (Qmax) were expressed as percentiles and compared to the Toguri value from normal children.

RESULTSBefore corrective operation, 12 of 17 patients (70.6%) produced a plateau urine flow curve. 5 patients (29.4%) produced a very low flow curve. The average maximum flow rate was (7.89 +/- 2.29) ml/s per second compared to Toguri values, 12 of 17 patients (70.6%) had a Qmax below the normal 5th percentile. After a "Two in One" urethroplasty, a hell-shaped curve was obtained in 10 patients (58.8%). The maximum flow rate was (11.30 +/- 3.01) mL/s per second. 7 of 17 patients (41.2%) had a Qmax above the normal 25th percentile, 8 patients (47.1%) had a Qmax between the normal 25th percentile and 5th percentile, only 2 patients (11.8%) had a Qmax below the 5th percentile.

CONCLUSIONSThe functional result of the "Two in One" urethroplasty is ideal. The maximum urine flow rate of the patients increases after the operation.

Child, Preschool ; Humans ; Hypospadias ; physiopathology ; surgery ; Male ; Treatment Outcome ; Urethra ; physiopathology ; surgery

Child ; Child, Preschool ; Female ; Flow Cytometry ; methods ; Humans ; Infant ; Male ; Thrombasthenia ; classification ; diagnosis

Four compounds were isolated from the 70% EtOH extract of Ardisia crispa by using various chromatographic techniques, including silica gel, ODS and semi-preparative HPLC. The structures of 1-4 were elucidated based on physicochemical properties and spectroscopic data. These compounds were defined as crispalactone A (1), (+)-pinoresinol (2), 3,5-dimethoxy-4- hydroxyphenol-1-O-β-D-glucopyranoside (3) and (+)-schizandriside (4). Compound 1 is a new γ-valerolactone derivative, and compounds 2-4 are firstly isolated from Ardisia crispa.

Objective To explore the protective effects of proanthocyanidins pretreatment on intestinal function after cerebral ischemia-reperfusion injury. Methods 24 Sprague-Dawley rats were randomly divided into sham group (group A, n=8), ischemia-reperfusion group (group B, n=8) and proanthocyanidins pretreatment group (group C, n=8). The model of cerebral ischemia-reperfusion injury in rats was established according to Longa's method. Group C was intraperitoneally injected with proanthocyanidins 10 mg/(kg ⋅ d), group A and group B were injected with normal saline for 5 consecutive days. 1 and 3 days after cerebral ischemia-reperfusion, ileum myoelectric slow wave and smooth muscle contractility, the activity of superoxide dismutase (SOD) and content of malondialdehyde (MDA) were measured, the content of the serum TNF-α was tested with ELISA kit, ileum tissues were tested with hematoxylin eosin (HE) staining and used for measuring the moisture content. Results Compared with group B 1 and 3 days after cerebral ischemia-reperfusion, the intestinal mucosa injury relieved, the intestinal mucosa score decreased (P<0.05) and the number of infiltrated inflammatory cell decreased in group C; the frequency of slow wave and contraction trended to increase (P>0.05), and the amplitude increased (P<0.05) in group C; the serum SOD activity increased (P<0.05), and the content of MDA and TNF-α decreased (P<0.01) in group C; the intestinal moisture content reduced (P<0.01) in group C. Conclusion Proanthocyanidins pretreatment can protect intestinal function from injury after cerebral ischemia-reperfusion.