Personalized digital simulation­assisted acetabular component implantation in revision hip arthroplasty.

The number of artificial total hip revision arthroplasties is increasing yearly in China, and >50% of these cases have acetabular defects. Accurately locating and quantifying the bone defect is one of the current challenges of this surgery. Thus, the objective of the present study was to simulate acetabular implantation with the aid of Mimics 17.0 software (Materialise NV) in patients with loosened acetabular prosthesis, to evaluate the 'ideal acetabular center' and the 'actual acetabular center' to guide the choice of prosthesis and surgical method. From January 2017 to June 2021, the present study included 10 hips from 10 patients [seven men (seven hips) and three women (three hips)]. In all patients, the Mimics software was applied to simulate the dislocation of the femoral prosthesis and acetabular prosthesis implantation before surgery; calculate the height difference between the 'ideal acetabular center' and the 'actual acetabular center' to assess the bone defect; confirm the size of the acetabular prosthesis, abduction angle, anteversion angle and bone coverage of the acetabular cup; and measure the intraoperative bleeding and postoperative follow-up Harris score of the hip joint. After statistical analysis, the present study revealed that digital simulation assistance could improve the accuracy of hip revision acetabular prosthesis implantation, reduce postoperative shortening of the affected limb, especially for surgeons with relatively little experience in hip revision surgery, and greatly reduce the occurrence of complications such as hip dislocation because of poor postoperative prosthesis position.

Circadian Clock REV-ERBs Agonist SR9009 Induces Synergistic Antitumor Activity in Multiple Myeloma by Suppressing Glucose-Regulated Protein 78-Dependent Autophagy and Lipogenesis.

Background: Proteasome inhibitors, such as bortezomib, have demonstrated efficacy in the therapeutic management of multiple myeloma (MM). However, it is important to note that these inhibitors also elicit endoplasmic reticulum stress, which subsequently triggers the unfolded protein response (UPR) and autophagy, which have been shown to facilitate the survival of tumor cells. The disruption of the circadian clock is considered a characteristic feature of cancer. However, how disrupted circadian clock intertwines with tumor metabolism and drug resistance is not clearly clarified. This work explores the antitumor effectiveness of bortezomib and the circadian clock agonist SR9009, elucidating their impact on glucose-regulated protein 78 (GRP78), the autophagy process, and lipogenesis. Methods: The antitumor effects of bortezomib and SR9009 were evaluated using human MM cell lines (RPMI8226 and U266) in vitro and in vivo nonobese diabetic/severe combined immunodeficient (NOD/SCID) murine xenograft MM model. The assessment of cell viability was conducted using the cell counting kit-8 (CCK8) method, whereas the measurement of cell proliferation was performed with the inclusion of EdU (5-ethynyl-2'-deoxyuridine). Apoptosis was assessed by flow cytometry. The cells were transduced using adenovirus-tf-LC3, which was labeled with dual fluorescence. Subsequently, confocal imaging was employed to observe and examine the autophagosomes. REV-ERBα knockdown leads to upregulation of ATG5 and BENC1 at the protein level with immunoblot. Changes in the expression levels of GRP78, LC3, stearoyl-CoA desaturase 1 (SCD1), and fatty acid synthase (FASN) were assessed through the utilization of quantitative real-time polymerase chain reaction (qRT-PCR) and western blotting. Results: Our results showed that both bortezomib and circadian clock REV-ERBs agonist SR9009 decreased MM viability, proliferation rate and induced an apoptotic response in a dose-dependent manner in vitro. However, the two differ greatly in their mechanisms of action. Bortezomib upregulated GRP78 and autophagy LC3, while circadian clock agonist SR9009 inhibited GRP78 and autophagy LC3. Combined SR9009 with bortezomib induced synergistic cytotoxicity against MM cells. REV-ERBα knockdown lead to upregulation of ATG5, BENC1 and significant upregulation of FASN, and SCD1. Mechanically, SR9009 inhibited the core autophagy gene ATG5 and BECN1, and two essential enzymes for de novo lipogenesis FASN and SCD1. SR9009 had synergistic effect with bortezomib and slowed down murine xenograft models of human MM tumor growth in vivo. Conclusions: Taken together, these results demonstrated that the circadian clock component REV-ERBs agonist SR9009 could inhibit GRP78-induced autophagy and de novo lipogenesis processes and had a synergistic effect with proteasome inhibitors in both in vitro and in vivo models of MM. Our findings shed light on how a disrupted circadian clock interacts with metabolic mechanisms to shape proteasome inhibitor drug resistance and suggest that SR9009 may be able to overcome the inherent drug resistance of proteasome inhibitors.

[Soil Heavy Metal Content, Pollution, and Influencing Factors in Typical Farming Area of Sichuan Basin].

In order to identify heavy metal contents, the pollution characteristics and influencing factors of soil in typical farming areas in the Sichuan basin were analyzed, with Jiangjin District of Chongqing City chosen as the study area. Two hundred and forty-seven topsoil samples were collected and analyzed using the Nemerow index (NPI), geographical information system (GIS), and geodetector method. The results demonstrated that: 1 the arithmetic means of Cd, Cu, and Zn in the topsoil were 1.22, 1.10, and 1.98 times that of the soil background values in western Chongqing, respectively. 2 The high-value areas of the six heavy metals were mainly distributed in the northern, western, and central Jiangjin district, whereas the low-value areas were distributed in the eastern and southern Jiangjin district. 3 The NPI showed that the polluted sample points accounted for 22.1% of the total sample points, indicating that the overall soil pollution was mainly safety and vigilance in general. The high value of NPI was distributed in Dingshan street in the northern Jiangjin district. 4 The explanatory power of stratum on the distribution of heavy metal contents in the topsoil was the strongest, followed by that of the topographic factor. The interaction effect of the stratum and topographic factors on the distribution of heavy metal content in soil was the strongest. The results showed that the stratum and topographic factors were the key factors affecting the distribution of soil heavy metal contents in the study area.

[Effect of Land Use/Land Cover Change on the Concentration of Se and Heavy Metals in Soils from a "Return Cropland to Forest" Area, Southwest China].

Human activities (land use) and environmental change (land cover change) affect the concentration of Se and heavy metals in soils. The implementation of the "Return Cropland to Forest (RCF)" ecological project has changed the land use and cover, which has provided an ideal experimental area for studying the effects of land use and cover change on selenium (Se) and heavy metals in the soil. In this study, 91 top soil samples from different land use and land cover types, including dry land, paddy land, natural forest land, and secondary forest land, were collected, and the contents of Se, heavy metals, and soil organic matter (SOM) and pH were analyzed. The results showed that:① the average values of ω(Se) (0.42×10-6), ω(As) (13.0×10-6), and ω(Sb) (1.03×10-6) were higher than the soil background values of western Chongqing. ② The concentrations of Se, Cd, Cr, Ni, Pb, and Zn in soils from secondary forest land were significantly higher than those from dry land soils, suggesting that the Se and heavy metals might have significantly increased in the surface soil after the implementation of the RCF ecological project. ③ The SOM was the key controlling factor for the enrichment and distribution of Se and heavy metals in the top soils. Our results indicated that the land use and land cover change would deeply impact the concentrations of Se and heavy metals in the top soils via influencing the soil properties, especially the SOM.

Spinocerebellar ataxia type 3 with dopamine-responsive dystonia: A case report.

BACKGROUND: Spinocerebellar ataxia type 3 (SCA3) is a rare neurodegenerative disease with high genetic heterogeneity. SCA3 mainly manifests as progressive cerebellar ataxia accompanied by paralysis of extraocular muscles, dysphagia, lingual fibrillation, pyramidal tract sign, and extrapyramidal system sign. However, it rarely has clinical manifestations similar to Parkinson-like symptoms, and is even rarer in patients sensitive to dopamine. We report a patient initially diagnosed with dopamine-responsive dystonia who was ultimately diagnosed with SCA3 by genetic testing, which was completely different from the initial diagnosis. CASE SUMMARY: A 40-year-old Chinese woman was admitted to hospital due to severe inflexibility. At the beginning of the disease, she presented with anxiety and sleep disorder. At the later stage, she presented with gait disorder, which was similar to Parkinson's disease. Her medical history was unremarkable, but her mother, grandmother, and uncle all had similar illnesses and died due to inability to take care of themselves and related complications. Laboratory and imaging examinations showed no abnormalities, but electromyography and electroencephalography revealed delayed somatosensory evoked potentials and slow background rhythm, respectively. Her symptoms fluctuated during the daytime, and we initially diagnosed her with dopamine-responsive dystonia. After treatment with low-dose levodopa, the patient's symptoms were significantly improved, but the final genetic diagnosis was SCA3. CONCLUSION: SCA3 has various clinical phenotypes and needs to be differentiated from Parkinson's syndrome and dopamine-responsive dystonia.

Where Skilled Nursing Facility Residents Get Acute Care: Is the Emergency Department the Medical Home?

OBJECTIVES: This study aimed to characterize the distribution of acute care visits among Medicare beneficiaries receiving skilled nursing facility (SNF) services. METHODS: We conducted a cross-sectional analysis of a 20% sample of continuously enrolled Medicare beneficiaries in the 2012 Chronic Condition Warehouse data set. Beneficiaries were grouped by the number of days of SNF services, and acute care visits were categorized as "before SNF," "during SNF," or "after SNF." RESULTS: Among the 10,717,786 Medicare beneficiaries analyzed, 384,312 (3.6%) had at least one SNF stay. DISCUSSION: Beneficiaries who received SNF services had a higher proportion of acute care visits made to emergency departments (EDs) than beneficiaries who did not receive SNF services. Also, a higher proportion of acute care visits were made to EDs by beneficiaries after a SNF stay in comparison to residents actively residing in a SNF. The acute care capabilities of SNFs and post-SNF transitions of care to the community setting are discussed.

The effectiveness of a self-efficacy-focused structured education programme on adults with type 2 diabetes: A multicentre randomised controlled trial.

AIMS AND OBJECTIVES: To evaluate the effectiveness of a self-efficacy-focused structured education programme on outcomes in adults with type 2 diabetes (T2DM) without insulin therapy. BACKGROUND: Structured education regarding metabolic control in T2DM adults without insulin therapy has not always been effective, and this lack of effectiveness might be due to overlooking self-efficacy. Whether a self-efficacy-focused structured education programme could improve metabolic and psychosocial outcomes for T2DM adults more effectively remains unknown. DESIGN: A multicentre parallel randomised controlled concealed label trial. METHODS: The study conducted in outpatients of four hospitals in China. A total of 265 T2DM adults without insulin therapy were randomly assigned to an intervention group of a self-efficacy-focused structured education programme (n = 133), or to a control group of routine education (n = 132). The differences in metabolic and psychosocial outcomes were investigated at baseline, three- and 6-month follow-ups. RESULTS: The primary outcome of A1C and the secondary outcomes of weight, body mass index, waist circumference, diastolic pressure, self-efficacy, self-management behaviours and knowledge improved significantly in the intervention group compared with the control group at 6-month follow-up. The differences in A1C between groups for patients with a low educational background at 6-month follow-up were significant. No significant differences were found in other secondary outcomes of systolic pressure, the blood lipid profile and diabetes distress between groups at 6-month follow-up. CONCLUSIONS: This programme can improve glycaemic control, weight control, diastolic pressure, self-efficacy, self-management behaviours and diabetes knowledge for T2DM adults. RELEVANCE TO CLINICAL PRACTICE: This self-efficacy-focused structured education programme is effective and can be incorporated into regular clinical care and led by trained staff (e.g. nurses), and it can be implemented in patients with low educational backgrounds.

The radiotherapy-sensitization effect of cantharidin: Mechanisms involving cell cycle regulation, enhanced DNA damage, and inhibited DNA damage repair.

BACKGROUND: Cantharidin is an inhibitor of protein phosphatase 2 A (PP2A), and has been frequently used in clinical practice. In our previous study, we proved that cantharidin could arrest cell cycle in G2/M phase. Since cells at G2/M phase are sensitive to radiotherapy, in the present study, we investigated the radiotherapy-sesitization effect of cantharidin and the potential mechanisms involved. METHODS: Cell growth was determined by MTT assay. Cell cycle was evaluated by flow cytometry. DNA damage was visualized by phospho-Histone H2A.X staining. Expression of mRNA was tested by microarray assay and real-time PCR. Clinical information and RNA-Seq expression data were derived from The Cancer Genome Atlas (TCGA) pancreatic cancer cohort. Survival analysis was obtained by Kaplan-Meier estimates. RESULTS: Cantharidin strengthened the growth inhibition effect of irradiation. Cantharidin drove pancreatic cancer cells out of quiescent G0/G1 phase and arrested cell cycle in G2/M phase. As a result, cantharidin strengthened DNA damage which was induced by irradiation. Moreover, cantharidin repressed expressions of several genes participating in DNA damage repair, including UBE2T, RPA1, GTF2HH5, LIG1, POLD3, RMI2, XRCC1, PRKDC, FANC1, FAAP100, RAD50, RAD51D, RAD51B and DMC1, through JNK, ERK, PKC, p38 and/or NF-κB pathway dependent manners. Among these genes, worse overall survival for pancreatic cancer patients were associated with high mRNA expressions of POLD3, RMI2, PRKDC, FANC1, RAD50 and RAD51B, all of which could be down-regulated by cantharidin. CONCLUSION: Cantharidin can sensitize pancreatic cancer cells to radiotherapy. Multiple mechanisms, including cell cycle regulation, enhanced DNA damage, and inhibited DNA damage repair, may be involved.

The rs75932628 and rs2234253 polymorphisms of the TREM2 gene were associated with susceptibility to frontotemporal lobar degeneration in Caucasian populations.

Polymorphisms of the triggering receptor expressed on myeloid cells 2 (TREM2) gene have been reported to be potentially associated with the risks of developing frontotemporal lobar degeneration (FTLD), with inconsistent conclusions. This study aims to comprehensively investigate the potential role of TREM2 variants in FTLD risks via a meta-analysis. We included a total of eight eligible articles. For TREM2 rs75932628, we observed a significantly increased FTLD risk in the models of T vs. C [Association Test, odds ratio (OR) = 2.43, 95% confidence interval (CI) = 1.43∼4.14, P = 0.001], CT vs. CC (OR = 2.27, 95% CI = 1.39∼3.71, P = 0.001), CT + TT vs. CC (OR = 2.27, 95% CI = 1.38∼3.71, P = 0.001), and Carrier T vs. C (OR = 2.26, 95% CI = 1.38∼3.69, P = 0.001). Similarly, we observed positive results for TREM2 rs2234253 in all of the genetic models (all OR > 1, P = 0.030). Nevertheless, we did not observe any statistical difference between the case and control groups in the pooled analyses of TREM2 rs142232675 and rs143332484 (all P > 0.05). Our findings identified the rs75932628 and rs2234253 polymorphisms of the TREM2 gene as risk factors for FTLD in Caucasian populations.

Updated meta-analysis of the role of APOE ε2/ε3/ε4 alleles in frontotemporal lobar degeneration.

We performed an updated meta-analysis to assess the role of the ε2/ε3/ε4 alleles of Apolipoprotein E gene (APOE) in frontotemporal lobar degeneration (FTLD). The relevant articles were retrieved from PubMed, CENTRAL, EMBASE and Web of Science databases, and 51 eligible case-control studies with 5123 cases and 20566 controls were selected after screening according to inclusion and exclusion criteria. Our analysis demonstrated that APOE ε4 was associated with increased FTLD risk in all genetic models (ε4 vs. ε3 allele, ε4 vs. ε2 allele, ε4 vs. ε2+ε3+ε4 allele, ε4 vs. ε2+ε3+ε4 carrier, ε4ε4 vs. ε3ε3, ε3ε4 vs. ε3ε3, ε3ε4+ε4ε4 vs. ε3ε3, ε4ε4 vs. ε3ε3+ε3ε4, all P < 0.01, odds ratio [OR] > 1). Subgroup analysis revealed significant association between APOE ε4 and FTLD (P < 0.01, OR > 1) for the Caucasian, Italian, population based (PB), P > 0.05 value of the Hardy-Weinberg Equilibrium (HWE), Newcastle-Ottawa scale score > 6, and behavioral variant frontotemporal dementia (bvFTD) subgroups. However, there was no significant association between the APOE ε2 allele and FTLD (P > 0.05) in most genetic models and sub-group analyses. Begg's and Egger's tests also revealed no publication bias, and sensitivity analysis showed that our data analysis was robust. Thus our meta-analyses suggest that APOE ε4 is a genetic risk factor in patients with FTLD.

Genomic characteristics of pancreatic squamous cell carcinoma, an investigation by using high throughput sequencing after in-solution hybrid capture.

Squamous cell carcinoma (SCC) of pancreas is a rare histotype of pancreatic ductal carcinoma which is distinct from pancreatic adenocarcinoma (AC). Although there are standard treatments for pancreatic AC, no precise therapies exist for pancreatic SCC. Here, we screened 1033 cases of pancreatic cancer and identified 2 cases of pure SCC, which were pathologically diagnosed on the basis of finding definite intercellular bridges and/or focal keratin peal formation in the tumor cells. Immunohistochemistry assay confirmed the positive expression of CK5/6 and p63 in pancreatic SCC. To verify the genomic characteristics of pancreatic SCC, we employed in-solution hybrid capture targeting 137 cancer-related genes accompanied by high throughput sequencing (HTS) to compare the different genetic variants in SCC and AC of pancreas. We compared the genetic alterations of known biomarkers of pancreatic adenocarcinoma in different pancreatic cancer tissues, and identified nine mutated genes in SCC of pancreas: C7orf70, DNHD1, KPRP, MDM4, MUC6, OR51Q1, PTPRD, TCF4, TET2, and nine genes (ABCB1, CSF1R, CYP2C18, FBXW7, ITPA, KIAA0748, SOD2, SULT1A2, ZNF142) that are mutated in pancreatic AC. This study may have taken one step forward on the discovery of potential biomarkers for the targeted treatment of SCC of the pancreas.

Differentiation of Rat Bone Marrow Mesenchymal Stem Cells Into Neuron-Like Cells In Vitro and Co-Cultured with Biological Scaffold as Transplantation Carrier.

BACKGROUND Autograft and allograft transplantation are used to prompt the regeneration of axons after nerve injury. However, the poor self-regeneration caused by the glial scar and growth inhibitory factors after neuronal necrosis limit the efficacy of these methods. The purpose of this study was to develop a new chitosan porous scaffold for cell seeding. MATERIAL AND METHODS The bone marrow mesenchymal stem cells (BMSCs) and tissue-engineered biomaterial scaffold compound were constructed and co-cultured in vitro with the differentiated BMSCs of Wistar rats and chitosan scaffold in a 3D environment. The purity of the third-generation BMSCs culture was identified using flow cytometry and assessment of induced neuronal differentiation. The scaffolds were prepared by the freeze-drying method. The internal structure of scaffolds and the change of cells' growth and morphology were observed under a scanning electron microscope. The proliferation of cells was detected with the MTT method. RESULTS On day 5 there was a significant difference in the absorbance value of the experimental group (0.549±0.0256) and the control group (0.487±0.0357) (P>0.05); but on day 7 there was no significant difference in the proliferation of the experimental group (0.751±0.011) and the control group and (0.78±0.017) (P>0.05). CONCLUSIONS Tissue engineering technology can provide a carrier for cells seeding and is expected to become an effective method for the regeneration and repair of nerve cells. Our study showed that chitosan porous scaffolds can be used for such purposes.

Establishment of a two-dimensional chiral HPLC system for the simultaneous detection of lactate and 3-hydroxybutyrate enantiomers in human clinical samples.

A two-dimensional chiral high-performance liquid chromatography system was established for simultaneous detection of lactate (LA) and 3-hydroxybutyrate (3HB) enantiomers in human clinical samples. d-LA is increased upon kidney damage but 3HB protected against kidney injury. Therefore, determining the concentrations of D,L-LA and D,L-3HB simultaneously would be useful for evaluating pathological conditions. LA and 3HB were pre-column-derivatized with the fluorescent reagent 4-(N-chloroformylmethyl-N-methylamino)-7-nitro-2,1,3-benzoxadiazole (NBD-COCl) at 60 °C for 15 min and separated in the first dimension with a capillary monolithic octadecylsilane column. The mobile phase consisted of 13% acetonitrile and 0.05% tirfluoroacetic acid in water. Chiralpak QD-AX and KSAACSP-001S enantioselective columns were used in the second dimension to separate LA and 3HB enantiomers, respectively. Mobile phases were mixed solutions of methanol and acetonitrile containing formic acid. The separation factors were 1.14 and 1.08, respectively. The detection limit of LA and 3HB enantiomers was 10 fmol/injection. This method was applied to human clinical samples; intra- and inter-day relative standard deviations of LA and 3HB enantiomers were, respectively, 1.04-3.25% and 1.61-5.12% in plasma, 9.19-11.2% and 4.60-5.89% in urine, and 7.12-8.90% and 2.86-6.97% in saliva. This novel analytical method is a powerful tool for investigating variations in LA and 3HB enantiomers under disease conditions.

[Protective effect of serum containing n-butanol fraction from Gynostemma pentaphyllum on NG108-15 cell apoptosis induced by Aß25.35 protein].

OBJECTIVE: To investigate the inhibition effect of serum containing n-butanol fractions from Gynostemma pentaphyllum on NG108-15 cell apoptosis induced by Aß25-35 protein in vitro. METHODS: The apoptosis of NG108-15 cells induced by Aß25-35 protein in vitro was evaluated by immunofluorescence and flow cytometry assay. The cellular Caspase-3 level during the apoptosis was determined by ELISA. RESULTS: The serum containing n-butanol fractions from Gynostemma pentaphyllum significantly inhibited the NG108-15 cells apoptosis induced by Aß25-35 protein in vitro,and decreased the cellular Caspase-3 level. CONCLUSION: The inhibition effect of n-butanol fractions from Gynostemma pentaphyllum on NG108-15 cell apoptosis induced by Aß25.35 protein is likely related to its potency on reducing of cellular Caspase-3 level.

[Effect of low density lipoprotein oxidizing in hyperlipidemia rats after treating with tetrahydrobiopterin].

OBJECTIVE: To explore the oxidative modification effect and its mechanism of low density lipoprotein (LDL) in hyperlipidemia (HL) rats after treating with tetrahydrobiopterin (BH4). METHODS: Fifty four 8-week-old male Wistar rats were used, these 54 rats were randomly divided into control group, high fat diet group (HL group), high fat diet and injected BH4 group (HL + BH4 group), and 18 in each group. The BH4 levels of blood fats and blood serum and its metabolites, the aortic reactive oxygen species, the end product malondialdehyde (MDA) and the LDL oxidation level were all determined by killing 6 experimental rats in each group at the first 8, 16, and 24 weeks of age respectively. RESULTS: Treating with BH4 after 8 and 16 weeks, there was no significant difference in serum lipids among three groups (P > 0. 05); but ROS and MDA decreased significantly (P < 0.01); compared with control and HL groups, the BH4 level of HL + BH4 group increased a lot (P < 0.01); compared with control group, the BH4 content reduced obviously in aortic homogenate of HL group (P < 0.01), but the total petrin levels (TB = BH4 + BH2 + B) had no significant difference (P > 0.05); the serum TBARS formation increased gradually with the increase of week-ages, but compared with HL group, the serum TBARS formation of HL + BH4 group reduced significantly (P < 0. 01). CONCLUSION: Treating with BH4 can reduced the LDL oxidation, the mechanism may be related to the correct of NOS uncoupling, the reduce of ROS generation and the decrease of LDL lipid peroxidation.

[Pretreatment of high-concentration acrylic acid wastewater by the multi-stage microaerobic biological fluidized bed reactor].

A multi-stage microaerobic biological fluidized bed reactor was used for the pretreatment of synthetic wastewater containing high concentration of acrylic acid (AA). The effect of influent load was investigated and the intermediate products of acrylic acid degradation were analyzed. It indicated that the removal rate of AA was above 95% with effluent acrylic acid less than 150 mg x L(-1) and COD removal rate of 15%-30%, under the following conditions: hydraulic retention time of 12 h, waste water temperature of 25 degrees C, influent acrylic acid concentration of 3 000-9 000 mg x L(-1), volume load of 6.0-18.0 kg x (m3 x d)(-1). The main intermediate products of acrylic acid degradation were acetic and propionic acids. The multi-stage microaerobic biological fluidized bed reactor can transform each 1.00 mol acrylic acid into 0.22 mol acetic acid and 0.36 mol propionic acid, and achieve the pretreatment of acrylic acid wastewater at high loads.

Involvement of Heme Oxygenase-1 Participates in Anti-Inflammatory and Analgesic Effects of Aqueous Extract of Hibiscus taiwanensis.

Anti-inflammatory effects of the aqueous extract of Hibiscus taiwanensis (AHT) were used in lipopolysaccharide (LPS-)stimulated mouse macrophage RAW264.7 cells and carrageenan (Carr-)induced mouse paw edema model. When RAW264.7 macrophages were treated with AHT together with LPS, a concentration-dependent inhibition of nitric oxide (NO), tumor necrosis factor (TNF-α), and prostaglandin E2 (PGE(2)) levels productions were detected. Western blotting revealed that AHT blocked protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), and elevated heme oxygenase-1 (HO-1), significantly. In the animal test, AHT decreased the paw edema at the 4th and the 5th h after Carr administration, and it increased the activities of catalase (CAT), superoxide dismutase (SOD), and glutathione peroxidase (GPx) in the paw tissue. We also demonstrated AHT decreased the NO, TNF-α, and PGE2 levels on the serum level at the 5th h after the Carr injection. Western blotting revealed that AHT decreased Carr-induced iNOS, and COX-2, and increased HO-1 expressions at the 5th h in the edema paw. These findings demonstrated that AHT has excellent anti-inflammatory activities in vitro and in vivo and thus it has great potential to be used as a source for natural health products.

Immunity induced by Staphylococcus aureus surface protein A was protective against lethal challenge of Staphylococcus aureus in BALB/c mice.

Staphylococcus aureus is the most common cause of hospital-acquired bacteremia. Due to emergence of antibiotic-resistant strains, these infections present a serious public health threat. In this study, to develop a broadly protective vaccine, we tested whether immune responses induced by several proteins associated with S. aureus toxicity could protect mice from lethal challenge with human clinical S. aureus isolate USA300. We found that the surface protein A (SasA) of S. aureus could protect mice from lethal challenge of the bacteria.

[Expression and neutralizing activity analysis of humanized single-chain antibody against anthrax protective antigen].

AIM: In order to reduce the human anti-mouse antibody (HAMA) response, anti-anthrax protective antigen scFv-5E1 has been humanized. METHODS: The authors have constructed a "humanized" antibody fragments by grafting the complementarity-determining regions (CDRs) of the murine anti-PA scFv-5E1 to human framework regions which showed maximal homology to the scFv-5E1 sequence. The humanized VH-5E1 and VL-5E1 DNA fragments were then joined by fusion PCR. The expression vectors named pET-hscFv-5E1 was constructed by cloning the humanized 5E1 scFv gene into the Nde I/EcoR I site. The transformed E.coli BL21(DE3) cells were propagated and induced by IPTG. RESULTS: Expression product was found as inclusion body with expected size by SDS-PAGE analysis. Soulable scFv showed binding ability to the protective antigen by ELISA analysis. The purified scFv showed good neutralizing activity in a cell model. CONCLUSION: The humanized scFv exhibited good binding and neutralizing activity. It lays a foundation for the development of therapeutic whole molecular antibody.

[Biomechanical properties of artery in spontaneously hypertensive rats after treating with tetrahydrobiopterin].

AIM: To explore the effect of remodeling and biomechanical properties after chronic treating with tetrahydrobiopterin (BH4) in spontaneously hypertensive rats. METHODS: The spontaneously hypertensive rat(SHR) were given with BH4 chronically. The opening angle in the zero-stress state , wall-to-lumen area ratios (W/L) of thoracic aorta and the relationship between pressure and diameter (P-D) of mesenteric artery were measured by computer image analysis in 4, 16, and 26 week-old respectively. RESULTS: Treating with BH4 chronically from 4 weeks-old in SHR, there was a significant decrease in morphometric parameters of the thoracic aorta and an increase in the zero-stress state of opening angle of elastic artery. The P-D curve of mesenteric artery moved upward. CONCLUSION: Treating with BH4 prevented the structure and function of artery from abnormal changing, including to attenuate the resistant vascular hypertrophy and recover the vascular elasticity and expansibility.