AIM:To investigate the effects of ox-LDL on TLR2 and TLR4 expression and production of TNF-?,IL-10,IL-12,NO and MDA in macrophages and to observe intervention effect of GW1929 in above procedure.METHODS:The mouse peritoneal macrophages were pretreated with ox-LDL(50 mg/L,100 mg/L)and GW1929(20 ?mol/L)respectively for 24 h.The concentrations of MDA,NO-2/NO-3,TNF-?,IL-10 and IL-12 in the culture fluid were detected.Flow cytometry was used to observe TLR2 and TLR4 expressions after the mouse peritoneal macrophages were pretreated with ox-LDL(50 mg/L)and GW1929(20 ?mol/L)respectively for 6 h,12 h,and 24 h.RESULTS:The concentrations of MDA,NO-2/NO-3,TNF-? and IL-10 in ox-LDL(50 mg/L,100 mg/L)group were higher than those in control and GW1929 group obviously,but the concentrations of above index in ox-LDL(50 mg/L,100 mg/L)+GW1929 group were lower than those in ox-LDL(50 mg/L,100 mg/L)group apparently.No IL-12 in every group was detected.Expressions of TLR-2 in ox-LDL+GW1929(6 h,12 h,24 h)group were lower than those in ox-LDL(6 h,12 h,24 h)group respectively.TLR-4 expressions in ox-LDL+GW1929(12 h)were lower than those in ox-LDL(12 h)apparently.CONCLUSION:ox-LDL up-regulates TLR2 and TLR4 expressions and promotes the production of ROX,NO,TNF-? and IL-10 in macrophages.GW1929 is capable of inhibiting the above ox-LDL effects.

Objective To investigate the effects of activator protein-1(AP-1)decoy oligodeoxynucleotides (ODNs)on the myocardial fibrosis induced by angiotension Ⅱ(Ang Ⅱ)in vitro.Methods CFs of neonatal Spra- gue-Dawley(SD)rats were isolated by trypsin digestion method.CFs were co-cultured with 10~(-7)mol/L Ang Ⅱ in the presence of different concentration of activator protein-1(AP-1)decoy ODNs or mutational AP-1 decoy ODNs for 24 h.Collagen synthesis was assessed by hydroxyproline and the mRNA expression of collagen Ⅰ,collagen Ⅲ.Results The concentration of hydroxyproline increased significantly after treated by 10~(-7)mol/L Ang Ⅱ;decoy ODNs on the range of 10-200 nmol/L dose dependently decrease synthesis of collagen;Ang Ⅱ stimulates mRNA expression of collagen Ⅲ(1.04?0.07 vs 1.63?0.071,n=3,P

OBJECTIVETo evaluate the efficacy of semiconductor low level laser irradiation for the treatment of postoperative exposure of hydroxyapatite orbital implants.

METHODS22 cases with postoperative exposure of hydroxyapatite orbital implants were divided into three groups according to the size of implants exposure. The exposure wound in the 3 groups was irradated with semiconductor low level laser 5 min per day for 5-15 days. The follow-up period ranged from 2 to 24 months.

RESULTSIn the group with less then 3 mm of exposure, the wound healed in 1 week after 5-10 days irradiation; in the group with implant exposure of 4-7 mm, the would healed in 1-2 weeks after 10-15 days irradiation; in the group with implant exposure of 8-10 mm, the would healed in 2-3 weeks after 10-15 days irradiation. Compared with the treatments of drugs and/or surgical repair, which was used for another 20 cases of exposure of hydroxyapatite orbital implants, semiconductor low level laser increased healing rate obviously in the groups with implant exposure of 4-7 mm and 8-10 mm (P = 0.019, 0.018).

CONCLUSIONSemiconductor low level laser has better effects than drugs and/or surgical repair for exposure of hydroxyapatite orbital implants.

Adolescent ; Adult ; Aged ; Child ; Durapatite ; therapeutic use ; Eye ; pathology ; radiation effects ; Female ; Follow-Up Studies ; Humans ; Low-Level Light Therapy ; methods ; Male ; Middle Aged ; Orbital Implants ; adverse effects ; Postoperative Complications ; etiology ; radiotherapy ; Semiconductors ; Treatment Outcome

OBJECTIVETo detect the incidence of the HFE gene C282Y and H63D mutations in patients with myelodysplastic syndromes (MDS) and aplastic anemia (AA), and analyze the relationship of these mutations with iron metabolism, and organs impairment from iron overload.

METHODSThe incidence of the C282Y and H63D mutations in 271 MDS, 402 AA patients and 1615 normal subjects was measured by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) combining with DNA sequencing. Iron metabolism parameters and iron overload indices were retrospectively compared between HFE gene mutation and unmutation groups in MDS and AA patients with no transfusion history.

RESULTSNo C282Y and C282Y/H63D compound mutation was detected in all the three groups. The incidence of H63D heterozygous and homozygous genotype did not significantly differ between AA cases and controls (9.7% vs 10.2%, 0.25% vs 0.24% respectively, both P > 0.05). The frequency of H63D heterozygous genotype in MDS patients was significantly lower than that in controls (4.1% vs 10.2%, P = 0.002). H63D homozygous was not found in MDS patients. In both MDS and AA patients with no RBC transfusion history, serum ferritin (SF), transferrin saturation value (TS), serum iron concentration (SI) were close to or higher than normal; and unsaturated iron-binding capacity (UIBC) value was significantly lower. There was no significant difference in SF, SI, TS values between HFE-mutation and -unmutation MDS patients. For AA patients, only the level of SI was significantly higher in HFE-mutant group than in -unmutation group [42.6 (24.6-60.4) micromol/L vs 32.0 (8.4-63.3) micromol/L, P = 0.011]. There was no significant difference in the values of liver enzyme, fasting blood sugar (FBS), abnormal electrocardiogram (ECG), peripheral blood indices between HFE-mutation and -unmutation MDS and AA groups (all P > 0.05).

CONCLUSIONThe distribution of C282Y and H63D mutations has ethnic and genetic disparity, the frequency in Chinese population is lower than that in Caucasian. It seems that MDS and AA patients are susceptible to iron overload, in the diseases itself and the mutations of HFE gene are not the major factor for iron overload in the patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anemia, Aplastic ; complications ; genetics ; Case-Control Studies ; Child ; Child, Preschool ; China ; Female ; Genotype ; Hemochromatosis Protein ; Histocompatibility Antigens Class I ; genetics ; Humans ; Iron ; blood ; Iron Overload ; etiology ; genetics ; Male ; Membrane Proteins ; genetics ; Middle Aged ; Mutation ; Myelodysplastic Syndromes ; complications ; genetics ; Young Adult

OBJECTIVETo investigate the inhibitory effects of AP-1 decoy oligodeoxynucleotides (ODNs) on angiotensin II (AngII)-induced proliferation and collagen synthesis in neonatal rat cardiac fibroblasts (CFs).

METHODSThe CFs of neonatal SD rats were cultured in serum-free medium for 24 h and stimulated with 10(-7) mol/L AngII in the presence of AP-1 decoy ODNs or mutational AP-1 decoy ODNs at varied concentrations. MTT assay was employed for quantitative evaluation of the CF proliferation. Collagen synthesis in the CFs was assessed with hydroxyproline, and the cell cycle distribution determined with flow cytometry (FCM).

RESULTSWith the increase of the concentration of AP-1 decoy ODNs, the absorbance at 490 nm (OD490) of the CFs decreased gradually as shown by MTT assay. Treatment with 100 or 200 nmol/L AP-1 decoy ODNs resulted in significantly lowered OD490 of the CFs as compared with that of AngII group. The concentration of hydroxyproline increased significantly after treatment with 10(-7) mol/L AngII in comparison with the control group (P<0.05). Hydroxyproline concentration in cells treated with 100 or 200 nmol/L AP-1 decoy ODNs was significantly lower than that in the 10(-7) mol/L AngII-treated cells. AP-1 decoy ODNs decreased the cell percentage in S phase and increased hydroxyproline concentration, but increased the percentage of cells in G0/G1 phase. AP-1 decoy ODNs at 100 and 200 nmol/L did not obviously affect AngII-induced CF proliferation and collagen synthesis (P<0.01).

CONCLUSIONAP-1 decoy can inhibit AngII-induced rat CF proliferation and collagen synthesis possibly by affecting the cell cycle distribution.

Angiotensin II ; pharmacology ; Animals ; Animals, Newborn ; Cell Cycle ; drug effects ; Cell Proliferation ; drug effects ; Cell Survival ; drug effects ; Cells, Cultured ; Collagen ; biosynthesis ; Dose-Response Relationship, Drug ; Fibroblasts ; cytology ; drug effects ; metabolism ; Flow Cytometry ; Mutation ; Myocardium ; cytology ; metabolism ; Oligodeoxyribonucleotides ; genetics ; pharmacology ; Rats ; Rats, Sprague-Dawley ; Transcription Factor AP-1 ; genetics

Protein tyrosine phosphatase (PTP) 1B is a potential target for the treatment of diabetes and obesity. Phosphotyrosine (pTyr) is the substrate for PTP1B dephosphorylation. Malonic acid moiety was used herein as a mimic of the phosphate group in pTyr, and novel malonic acid derivatives 1-7 were designed, synthesized and evaluated as PTP1B inhibitors. Results from enzymatic assays indicated that compounds 3 and 4 exhibited potent inhibition against human recombinant PTP1B with IC50 values of 7.66 and 1.88 micromol x L(-1), respectively.
Drug Design ; Enzyme Inhibitors ; chemical synthesis ; chemistry ; pharmacology ; Humans ; Inhibitory Concentration 50 ; Malonates ; chemical synthesis ; chemistry ; pharmacology ; Molecular Structure ; Protein Tyrosine Phosphatase, Non-Receptor Type 1 ; antagonists & inhibitors ; metabolism ; Structure-Activity Relationship

BackgroundFat grafting technologies are popularly used in plastic and reconstructive surgery. Due to its size limitation, it is hard to directly inject untreated fat tissue into the dermal layer. Nanofat, which was introduced by Tonnard, solves this problem by mechanically emulsifying fat tissue. However, the viability of the cells was greatly destroyed. In this study, we reported a new method by "gently" digesting the fat tissue to produce viable adipocytes, progenitors, and stromal stem cells using collagenase I digestion and centrifugation. This was named "Vivo nanofat".

MethodsHuman liposuction aspirates were obtained from five healthy female donors with mean age of 28.7 ± 5.6 years. Colony-forming assay, flow cytometry analysis, and adipogenic and osteogenic induction of the adherent cells from the Vivo nanofat were used to characterize the adipose mesenchymal stem cells (MSCs). To investigate in vivo survival, we respectively injected Vivo nanofat and nanofat subcutaneously to the back of 8-week-old male BALB/c nude mice. Samples were harvested 2 days, 2 weeks, and 4 weeks postinjection for measurement, hematoxylin and eosin staining, and immunostaining.

ResultsOur results showed that the Vivo nanofat contained a large number of colony-forming cells. These cells expressed MSC markers and had multi-differentiative potential. In vivo transplantation showed that the Vivo nanofat had lower resorption ratio than that of nanofat. The size of the transplanted nanofat was obviously smaller than that of Vivo nanofat 4 weeks postinjection (0.50 ± 0.17 cm vs. 0.81 ± 0.07 cm, t = -5783, P = 0.01).

ConclusionVivo nanofat may serve as a cell fraction injectable through a fine needle; this could be used for cosmetic applications.

Objective A variety of miRNAs have been found to be involved in the occurrence and development of colorectal cancer. This paper aim to investigate the clinical and biological relevance of miR-217 and the pathway by which miR-217 may be involved in progression in colorectal cancer. Methods According to the diameter of tumor, the tumor was divided into tumors>5 cm(n=22) and tumors≤5 cm(n=28); 18 cases of colorectal cancer I-II specimens and 32 of III-IV patients; according to median expression of miR-217, the specimens were divided into high expression group and low expression group. The expression of miR-217 in 50 colon cancer patients’ carcinoma and adjacent tissues was determined by qRT-PCR. Cells were grouped: overexpression control Group (transfected control mimic) and miR-217 overexpression group(transfected miR-217 mimic), low expression control group((transfected control inhibitor) and miR-217 low expression group(transfected miR-217 inhibitor), miR-217&ERK1/2 low expression group(transfected miR-217 inhibitor+U0126). MiR-217 and HCT116 cells were over expressed in SW480 cells,and miR-217 was knocked down. A series of biological function assays were performed to assess cell viability (cck-8 assay), clony formation ability (clony formation assay), proliferation (edu assay), Changes in ERK1/2 expression were measured at protein level, and the relationship between miR-217 and ERK1/2 in colorectal cancer cells was explored by relevant rescue experiments. Results Compared with colorectal adjacent noncancerous tissues, the expression of miR-217 was significantly decreased in carcinoma tissues(-1.360±0.645 vs 2.244±0.168, P<0.01); the expression of miR-217 in tumors with a diameter >5cm was significantly lower than that of tumors with a diameter ≤5cm(-1.718±0.272 vs -0.587±0.288, P<0.01); the expression of miR-217 in stage I-II colorectal cancer tissues was significantly higher than that stage III-IV(-0.413±0.330 vs -01.463±0.230, P<0.05); the expression of miR-217 in the miR-217 overexpression group was significantly higher than miR-217 overexpression control group(15.120±0.522 vs 1.004±0.003, P<0.01), and the number of clone formation was significantly less than that of the overexpression control group(199.30±15.62 vs 439.70±18.91, P<0.01) . In HCT116 cells, the expression of miR-217 in the miR-217low expression group was significantly lower than miR-217 low expression control group(0.2070±0.021 vs 1.006±0.003, P<0.01), and the number of clone formation was significantly higher than that control group(237.30±12.14 vs 117.00±7.00, P<0.01) . When miR-217 overexpressed in SW480, the protein expression of ERK1/2 decreased; when miR-217 was inhibited in HCT116, the protein expression of ERK1/2 increased. The number of colons in ERK1/2 low expression group was significantly lower than that miR-217&ERK1/2 low expression group(221.70±12.73 vs 108.00±5.51) , the difference was statistically significant(P<0.01). Conclusion Low expression of miR-217 is observed in both colorectal cancer tissues and cells, and miR-217 can affect tumor cell proliferation in the progression of colorectal cancer partly by inhibiting the expression of ERK1/2.

In order to establish the probabilistic risk assessment method for heavy metals and harmful elements in line with the characteristics of traditional Chinese medicine (TCM) and provide guidance for the safe use of TCM, the contents of lead (Pb), cadmium (Cd), arsenic (as), mercury (Hg) and copper (Cu) in 21 batches of Plantago asiatica L. were determined by inductively coupled plasma mass spectrometry (ICP-MS). By the comprehensive use of investigation of TCM consumption pattern and Monte Carlo simulation technology, the non-carcinogenic and carcinogenic health risks of heavy metals and harmful elements in TCM were assessed by hazard quotient (HQ) and cancer risk (CR), respectively. The greatest risk contributors were screened through sensitivity analysis. The results revealed that the mean contents of Pb, Cd, As, Hg and Cu in Plantago asiatica L. were 9.01, 0.28, 3.83, 0.01 and 12.82 mg·kg^-1, respectively. The P95, P99 and maximum values of HQ for males were 1.41, 2.83 and 10.97 mg·kg^-1, respectively. The P95, P99 and maximum values of HQ for females were 1.27, 2.63 and 9.80 mg·kg^-1, respectively. The P95, P99 and maximum values of HI for males were 1.44, 2.85 and 11.0 mg·kg^-1, respectively. The P95, P99 and maximum values of HI for females were 1.29, 2.66 and 10.0 mg·kg^-1, respectively. The P99 and maximum values for CR_As and total carcinogenic risk (TCR) were greater than 10^-4 for both men and women. The risk assessment results indicated that the non-carcinogenic and carcinogenic health risks of high exposure population caused by arsenic exposure are needed to be concern. The results of sensitivity analysis showed that the exposure frequency of TCM and the arsenic concentrations in Plantago asiatica L. were the main risk contributors. Based on Monte Carlo simulation technology and considering the characteristics of TCM, this study puts forward the first example of probabilistic risk assessment of heavy metals and harmful elements in Chinese herbal medicine, which provides a novel perspective for health risk assessment of heavy metals in TCM.

Infectious bone defect is bone defect with infection or as a result of treatment of bone infection. It requires surgical intervention, and the treatment processes are complex and long, which include bone infection control,bone defect repair and even complex soft tissue reconstructions in some cases. Failure to achieve the goals in any step may lead to the failure of the overall treatment. Therefore, infectious bone defect has been a worldwide challenge in the field of orthopedics. Conventionally, sequestrectomy, bone grafting, bone transport, and systemic/local antibiotic treatment are standard therapies. Radical debridement remains one of the cornerstones for the management of bone infection. However, the scale of debridement and the timing and method of bone defect reconstruction remain controversial. With the clinical application of induced membrane technique, effective infection control and rapid bone reconstruction have been achieved in the management of infectious bone defect. The induced membrane technique has attracted more interests and attention, but the lack of understanding the basic principles of infection control and technical details may hamper the clinical outcomes of induced membrane technique and complications can possibly occur. Therefore, the Chinese Orthopedic Association organized domestic orthopedic experts to formulate An evidence-based clinical guideline for the treatment of infectious bone defect with induced membrane technique ( version 2023) according to the evidence-based method and put forward recommendations on infectious bone defect from the aspects of precise diagnosis, preoperative evaluation, operation procedure, postoperative management and rehabilitation, so as to provide useful references for the treatment of infectious bone defect with induced membrane technique.