Animals ; Cell Proliferation ; drug effects ; Cells, Cultured ; Collagen ; biosynthesis ; Cricetinae ; Cricetulus ; Fibroblasts ; cytology ; drug effects ; metabolism ; Macrophages ; drug effects ; Nanostructures ; Silicon Dioxide ; pharmacology

BACKGROUND:Core decompression may provide insufficient support for the subchondral bone in the treatment of early avascular necrosis of the femoral head and increase the risk of fracture and colapse. Quadratus femoris implantation cannot only provide good biological support, but also promote the revascularization at necrotic regions, thus repairing the necrosis of femoral head. OBJECTIVE: To compare the clinical effects of core decompression with bone paste implantation and core decompression with quadratus femoris implantation on early and middle-stage avascular necrosis of the femoral head. METHODS:Eighty-three patients with early avascular necrosis of the femoral head (92 hips) admitted at the Department of Orthopedics, the First Affiliated Hospital of Zhengzhou University, China, from January 2009 to January 2012 were enroled and divided into groups of core decompression with bone paste implantation (46 cases, 49 hips) and core decompression with quadratus femoris implantation (37 cases, 43 hips) that were respectively injected with bone meal and autogenous bone and osteoinductive materials. RESULTS AND CONCLUSION: Al involved patients were folowed up. After 1 year of treatment, Harris scores in the two groups were both increased (P < 0.05). But the Harris score of core decompression with bone paste implantation group was lower than that of core decompression with quadratus femoris implantation group (P < 0.05). After 3 years of treatment, X-ray scores in the core decompression with quadratus femoris implantation group were significantly higher than those in the core decompression with bone paste implantation group (P < 0.05). These findings indicate that compared with core decompression with bone paste implantation, core decompression with quadratus femoris implantation is better to prevent femoral head colapse, improve hip function and delay the process of osteonecrosis of the femoral head.

[Objective] To observe the effect of Xiangdan injection with the actions of activating blood and removing stasis for the treatment of acute tubular necrosis (ATN) after kidney transplantation. [Methods] Twenty-six patients with ATN after kidney transplantation were randomized into groups A and B. The two groups were treated with triple immuno-suppressive agents of cyclosporin A, prednisone and mycophenolic acid, with medicines for protecting liver function and gastric mucosa, and with preventive measures from infection. Additionally, group A (n = 16) was treated with Xiangdan injection, while group B ( n = 10) was not. After treatment, time for hemodialysis, renal function and renal flow resistance index of the transplanted kidney were compared between the two groups after the occurrence of ATN. [Results] Compared with group B, time for hemodialysis was shortened, the decrease of serum creatine (SCr) in the transplanted kidney quickened, and renal flow resistance index (RI) of the transplanted kidney decreased (P 0.05). [Conclusion] Xiangdan injection with the actions of activating blood and removing stasis can promote the early recovery of ATN after kidney transplantation.

Objective To investigate the mechanism of an inhibitor of NADPH oxidases,diphenylene iodonium (DPI),in preventing radiation-induced lung injury.Methods Totally 48 adult SD male rats were randomly classified into 4 groups:control group (C),radiation group (R),radiation plus DPI group (R + D) and DPI group (D).The radiation induced pulmonary injury model was preformed by using 6 MV X-rays to deliver 8 Gy per day for 5 consecutive days with 40 Gy in total to the thorax of each animal.Rats in R + D group were subcutaneously administered with 0.02％ DPI (1 mg/kg) at 1 h prior to radiation while rats in D group received the same dose of DPI without radiation.DPI was given from 3 d before radiation to 30 d after the first radiation.Rats in C and D groups received the same dose of saline.Animals were sacrificed at 1 month and 6 months after radiation,respectively.The lungs were removed and processed for HE and Masson staining,hydroxyproline content measurement,and TGF-β1 immunohistochemical detection.Results At 1 month post-radiation,rats in R group showed typical alveolitis,the level of hydroxyproline was (0.69 ± 0.05) μg/mg,and the positive area of TGF-β1 expression was (39.97 ± 0.90) ％,while the level of hydroxyproline in R + D group was (0.55 ± 0.03) μg/mg and the positive area of TGF-β1 expression was(33.83 ± 1.55) ％,rats in R + D group showed less severe alveolitis compared with R group(t =5.32,5.93,P ＜0.05).At 6 months post-radiation,rats in R group showed typical lung fibrosis with hydroxyproline level of (1.04 ±-0.02) μg/mg and TGF-β1 expression of (37.80 ± 0.85) ％,whereas the hydroxyproline level in R + D group was (0.85 ± 0.02) μg/ mg,the TGF-β1 expression was(23.93 ± 1.16)％,rats in R + D group showed moderate lung fibrosis(t =15.77,16.68,P ＜ 0.05),rats in C and D group had no noticeable changes.Conclusions Diphenylene iodonium could prevent radiation-induced lung injury by reducing the level of hydroxyproline and the expression of TGF-β1.

BACKGROUND: Vibrio vulnificus inside the body could activate the NF-κB signaling pathway and initiate the inflammatory cascade. The lung is one of the earliest organs affected by sepsis associated with acute lung injury. High mobility group protein B1 (HMGB1) is an important late-acting pro-inflammatory cytokine involving in the pathophysiology of sepsis. It is also involved in the injury process in the lung, liver and intestine. There has been no report on the involvement of HMGB1 in Vibrio vulnificus sepsis-induced lung injury. METHODS: Sixty rats were randomly divided into a normal control group (group A,n=10) and a Vibrio vulnificus sepsis group (group B,n=50). Sepsis was induced in the rats by subcutaneous injection of Vibrio vulnificus (concentration 6×108 cfu/mL, volume 0.1 mL/100g)) into the left lower limbs. The rats in group B were sacrificed separately 1, 6, 12, 24, and 48 hours after the infection. Their lungs were stored as specimens, lung water content was measured, and lung pathology was observed under a light microscope. The expressions of the HMGB1 gene and protein in the lungs were detected by RT-PCR and Western blot. Data were analyzed with one-way analysis of variance (ANOVA) and the LSD method for pair-wise comparison between the two groups.P<0.05 was considered statistically significant. RESULTS: Compared to group A (0.652±0.177), HMGB1 mRNA expression in the lungs of group B was significantly higher at 0 hour (1.161±0.358,P=0.013), 24 hours (1.679±0.235,P=0.000), and 48 hours (1.258±0.274,P=0.004) (P<0.05), and peaked at 24 hours. Compared to group A (0.594±0.190), HMGB1 protein expression at 6 hours (1.408±0.567,P=0.026) after infection was significantly increased (P<0. 05), and peaked at 24 hours (2.415±1.064,P=0.000) after infection. Compared to group A (0.699±0.054), lung water content was significantly increased at 6 hours (0.759±0.030,P=0.001),12 hours (0.767±0.023,P=0.000), 24 hours (0.771±0.043,P=0.000) and 48 hours (0.789±0.137,P=0.000) after infection (P<0.05). Compared to group A, pathological changes at 12 hours in group B indicate marked pulmonary vascular congestion, interstitial edema and inflammatory infiltration. Alveolar cavity collapse and boundaries of the alveolar septum could not be clearly identified. CONCLUSION:Vibrio vulnificus sepsis can lead to injury in rat lungs, and increased HMGB1 expression in lung tissue may be one of the mechanisms for injury from Vibrio vulnificus sepsis.

Objective To explore the characteristic of fractional amplitude of low frequency fluctuation (fALFF) and the relationship with the severity of depression, suicidal ideation and suicide risk in depression patients with suicidal ideation with resting-state functional magnetic resonance imaging (rs-fMRI). Methods Resting state functional magnetic resonance imaging maps were conducted using fractional amplitude of low frequency fluctuation (fALFF) in 52 depression patients (30 with suicidal ideation and 22 without) and 21 healthy controls (HCs). The severity of depression was evaluat-ed by using Hamilton Depression scale(HAMD). The suicidal ideation, the suicide risk in depression patients with sui-cidal ideation were both assessed by the Beck Scale for Suicide Ideation. The correlation between the fALFF value and the score of HAMD and the Beck Scale for Suicide Ideation was analyzed. Results MRI revealed significant differences in fALFF in the left superior/middle occipital gyrus and the right middle/inferior occipital gyrus (P<0.05, AlphaSim cor-rected)between depression patients with suicidal ideation and the HCs. Compared to the HCs, depression patients with-out suicidal ideation showed a higher fALFF in the left middle occipital gyrus (P<0.05, AlphaSim corrected). MRI re-vealed significant differences in fALFF in the left middle occipital gyrus (P<0.01, AlphaSim corrected)and the right mid-dle occipital gyrus (P<0.01, AlphaSim corrected) between depression patients with suicidal ideation and without. The fALFF of left middle occipital gyrus (r=0.366, P=0.046) and right middle occipital gyrus (r=0.513, P=0.004) were posi-tively correlated with the scores of HAMD, respectively whereas were not correlated with suicidal ideation and suicide risk. Conclusions Depression patients with suicidal ideation have an abnormal spontaneous activity in their left and right middle occipital gyrus. The increased activity in these brain areas are probably associated with the severity of de-pression whereas are not associated with suicidal ideation or suicide risk.

BACKGROUND AND OBJECTIVEChemotherapy is the main treatment for colon cancer, while multidrug-resistance is the main reason for chemotherapy failure and tumor relapse. This study was to establish two oxaliplatin-resistant colon cancer cell lines and evaluate their biological characteristics.

METHODSOxaliplatin-resistant colon cancer cell lines SW620/L-OHP and lovo/L-OHP were established in vitro by continuous exposure to oxaliplatin (L-OHP) of low and gradually increased concentration. Growth curve, cross-resistance and resistance index of the oxaliplatin-resistant cell lines to various anti-cancer agents were determined by CCK8 assay. The expressions of P-glycoprotein (P-gp), multidrug-resistance protein 1 (MRP1) and MRP2 were detected by Western blot. Cell cycle distribution as well as the expression of CD133 and CD44 were measured by flow cytometry.

RESULTSIt took 10 months to establish the SW620/L-OHP and LoVo/L-OHP cell lines with stable resistance to oxaliplatin. Cross-resistance to 5-fluorouracil, etoposide, cisplatin, vincristine and epirubicin but not to paclitaxel was observed. Longer doubling time, higher proportion of cells in G(0)/G(1) phase and lower proportion in G(2)/M phase were observed in the two oxaliplatin-resistant cell lines compared with their parental cell lines. The expression of MRP2 in the oxaliplatin-resistant cells was up-regulated, while those of P-gp and MRP1 had no significant change. CD133 was overexpressed while CD44 level remained unchanged in SW620/L-OHP and LoVo/L-OHP cells.

CONCLUSIONSSW620/L-OHP and LoVo/L-OHP cell lines show a typical and stably resistant phenotype and may be used as research models.

AC133 Antigen ; ATP-Binding Cassette, Sub-Family B, Member 1 ; metabolism ; Antibiotics, Antineoplastic ; pharmacology ; Antigens, CD ; metabolism ; Antimetabolites, Antineoplastic ; pharmacology ; Antineoplastic Agents ; pharmacology ; Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Cycle ; Cell Line, Tumor ; Cisplatin ; pharmacology ; Colonic Neoplasms ; metabolism ; pathology ; Drug Resistance, Multiple ; Drug Resistance, Neoplasm ; Epirubicin ; pharmacology ; Etoposide ; pharmacology ; Fluorouracil ; pharmacology ; Glycoproteins ; metabolism ; Humans ; Hyaluronan Receptors ; metabolism ; Multidrug Resistance-Associated Proteins ; metabolism ; Organoplatinum Compounds ; pharmacology ; Peptides ; metabolism ; Vincristine ; pharmacology

OBJECTIVEThe effect of mesenchymal stem cells (MSCs) transplantation is poor because of the harsh environment post infarction. Our previous studies have proven that Statins could enhance the implanted bone marrow MSCs survival, but the exact mechanism remained to be clarified. We hypothesized that atorvastatin (Ator) could protect MSCs from hypoxia and serum-free (H/SF) induced apoptosis and investigated the potential mechanisms.

METHODSChinese mini-swine's bone marrow derived MSCs were cultured in vitro and exposed to hypoxia and H/SF, Ator of various concentrations (0.001 - 10 µmol/L), AMPK inhibitor-compound C (CC), PI3K inhibitor-LY294002 (LY), Ator + CC and Ator + LY. Cell apoptosis was assessed using Annexin V/Prospidine Iodine kit by flow cytometry. Phosphorylation of AMPK, Akt, endothelial nitric oxide synthase (eNOS) level and phosphorylation were tested with Western blot. Real Time-PCR was performed to analyze the gene expression of AMPK, Akt and eNOS.

RESULTSMSCs apoptosis in Ator (0.01 - 10 µmol/L) treated H/SF groups was significantly reduced compared with H/SF group (1.94% - 6.10% vs. 10.94%, P < 0.01 or 0.05). Apoptosis was higher in Ator + CC group than in 1 µmol/L Ator group (4.94% ± 0.98% vs. 2.59% ± 0.84%, P < 0.01) and similar between Ator + LY and 1 µmol/L Ator group (2.02% ± 0.45% vs. 2.59% ± 0.84%, P > 0.05). The gene expressions of AMPK, Akt and eNOS were significantly upregulated in atorvastatin treated groups. Meanwhile, phosphorylation of AMPK and eNOS increased in MSCs treated with atorvastatin (P < 0.01 or 0.05). Phosphorylation of eNOS significantly correlated with AMPK phosphorylation (r = 0.599, P = 0.004), but not with Akt phosphorylation (P = 0.263).

CONCLUSIONSAtorvastatin can protect MSCs from H/SF induced apoptosis through AMPK pathway, which resulting in activation of eNOS.

Adenylate Kinase ; metabolism ; Animals ; Apoptosis ; drug effects ; Atorvastatin Calcium ; Bone Marrow Cells ; cytology ; metabolism ; Culture Media, Serum-Free ; Heptanoic Acids ; pharmacology ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Nitric Oxide Synthase Type III ; metabolism ; Phosphatidylinositol 3-Kinases ; metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt ; metabolism ; Pyrroles ; pharmacology ; Signal Transduction ; Swine

OBJECTIVETo diagnose a Chinese benign familial neonatal convulsions (BFNC) family at the level of gene and investigate its molecular pathogenesis.

METHODSAll family members were studied by clinical examinations and linkage analysis. Mutation analysis of KCNQ2 gene was made by means of polymerase chain reaction (PCR)-direct sequencing and PCR-single strand conformation polymorphism (SSCP) in the proband, 16 family members and 72 unrelated normal individuals.

RESULTSLinkage analysis hinted the linkage of BFNC to KCNQ2, while the linkage to KCNQ3 was excluded. Mutation 1931delG of KCNQ2 gene was found in the proband by DNA-direct sequencing. The same SSCP variant as the proband's was showed in the rest affected members of this family but not in the unaffected members of this family and all of the 72 unrelated normal individuals.

CONCLUSION1931delG of KCNQ2 gene can cause BFNC in China and is novel mutation. The combination of linkage analysis and gene analysis is useful for gene diagnosis.

Epilepsy, Benign Neonatal ; genetics ; Female ; Genetic Linkage ; Humans ; Infant, Newborn ; KCNQ2 Potassium Channel ; KCNQ3 Potassium Channel ; Mutation ; Potassium Channels ; chemistry ; genetics ; Potassium Channels, Voltage-Gated

Objective To explore the effects and mechanism of electroacupuncture(EA)on denervation-induced atrophy. Methods A total of 21 male Sprague-Dawley rats were divided into sham group(n=7),model group(n=7)and EA group (n=7).The latter two groups were cut off their right sciatic nerve.Since one day after modeling,EA group accept-ed electroacupuncture at right Zusanli(ST36)and Huantiao(GB30)for eight weeks.Then,the gastrocnemius of all the rats were obtained,and measured the wet mass ratio.Cross-sectional area(CSA)and fiber diameter were measured after HE staining. The expression of autophagy-related gene ULK1, Atg13, Beclin1, Atg14, Atg7, Atg12,Atg5 and Atg16L1 were tested with reverse transcription real-time quantitative polymerase chain reaction. Results Compared with the sham group,the wet mass ratio,CSA and fiber diameter of gastrocnemius were lower signifi-cantly in the model group and EA group(P<0.001),and they were more in EA group than in the model group(P<0.05).Compared with the sham group,the mRNA expression of ULK1,Atg13,Beclin1,Atg14,Atg7,Atg12,Atg5 and Atg16L1 was more significantly in the model group (P<0.001), and they decreased in EA group compared with those of the model group(P<0.05). Conclusion Electroacupuncture can inhibit the overexpression of autophagy-related gene in denervated rats,which may steady skeletal muscle cells to delay atrophy.