[Potential application of improved hard tissue section technique in the clinical pathological diagnosis of bone and bone marrow].

Objective: To discuss the application value of hard tissue section in the clinicopathology diagnosis. Methods: From March 2021 to December 2021, bone slices of 19 patients (1 patient with osteochondroma, 2 patients with chondrosarcoma, 4 patients with osteosarcoma, 2 patients with fibrous dysplasia, 2 patients with bone metastasis from thyroid papillary carcinoma, 2 patients with osteomyelitis, 4 patients with giant cell tumor of bone, 2 patients with Ewing sarcoma) and 16 hemopathy patients were collected from the Department of Pathology, Shanghai Sixth People's Hospital. Of the osteopathy patients, there were 14 male and 5 female with a median age of 31 (10-66) years. Meanwhile, there were 7 male and 9 female with a median age of 28 (16-65) years among these hemopathy patients. Thirty-five cases were treated with modified hard tissue slicing technique and paraffin embedding technique, respectively. The advantages and disadvantages of the two methods for clinical diagnosis of bone disease were compared by Hematoxylin-Eosin staining (H&E staining), immunohistochemical staining (IHC), fluorescence in situ hybridization (FISH) and Sanger sequencing. Results: The improved resin-embedded method showed better histological morphology and cell structure. Besides, the expression of Ki67, SATB2, CD34, SMA, CD68,MPO,CD4 and CD33 in immunohistochemical staining in bone tissues which were embedded in resin were more clear in the accurate positive localization than those using paraffin-embedded. MDM2 of FISH exhibited a higher fluorescence intensity and more accurate location. Meanwhile, both methods treated with Sanger sequencing met the requirements of DNA purity and mutation detection. Conclusion: The improved hard tissue section method is simple and short time-consuming, which is suitable for optimizing the clinical bone and bone marrow pathological diagnosis process.

A New Mutation Genotype of K218T in Myosin-5 Confers Resistance to Phenamacril in Rice Bakanae Disease in the Field.

In 2017 and 2018, a total of 294 Fusarium fujikuroi isolates were collected from bakanae-diseased rice plants in Jinhua, Shaoxing, and Jiaxing in Zhejiang Province, China. Phenamacril sensitivity of these isolates was determined by the 50% effective concentration value or minimum inhibitory concentration methods. Our results indicated that the phenamacril resistance frequency of F. fujikuroi increased from 18% in 2017 to 47% in 2018, and rice plants infected with F. fujikuroi-resistant isolates could not be protected effectively with 50 mg/liter of phenamacril. Phenamacril-resistant F. fujikuroi isolates obtained from rice fields showed stable resistance, because their fitness levels (i.e., mycelial growth, sporulation, and pathogenicity) were similar to the phenamacril-sensitive isolates. In addition to the point mutation at codon 219 in the myosin-5 gene that conferred resistance to phenamacril, our results also showed another point mutation at codon 218 (AAG→ACG) in myosin-5 that also conferred resistance to phenamacril. In this study, we found rapid development and persistence of diversified genotypes of phenamacril resistance, highlighting the importance of proper use of phenamacril in rice fields. Our results may also help researchers develop new fungicides or new control strategies using combinations of different fungicides in the control of phenamacril-resistant F. fujikuroi isolates.

[Study of local application of CpG-oligodeoxynucleotide combined with 4-1BB monoclonal antibody to treat hepatoma- bearing mice].

Objective: To investigate the curative effect of local application of CpG-oligodeoxynucleotide (CpG-ODN) combined with 4-1BB monoclonal antibody in hepatoma-bearing mice, and to evaluate the effect of 4-1BB monoclonal antibody on CpG-ODN immunotherapy. Methods: H22 single cell suspension was injected subcutaneously into the axilla and four limbs of the BALB/c male mice to establish a tumor-bearing mice model. After 7 days, 30 mice with corresponding tumor-bearing volume were screened and randomly divided into model control group, CpG group and CpG+4-1BB group, and the drug was injected into the tumors of left lower extremity. The same batch of normal mice was selected as normal control group. Survival of mice was recorded. Tumor-bearing volume and organ index were calculated. Serum levels of interleukin (IL) - 12 and interferon (IFN) gamma and spleen CD8(+)T lymphocyte ratio were measured. The measurement data were analyzed by analysis of variance. The survival rate of each group of mice was analyzed by log-rank test. Results: Mice in the model control group with tumor-bearing volume had a sustained growth before the execution. CpG group and the CpG+4-1BB group [(976.08 ± 29.55) mm(3), (47.25 ± 0.93) mm(3))] tumor-bearing volume was decreased than model group [(1 336.52 ± 39.40) mm3] (F = 5 329.273, P < 0.05). CpG+4-1BB group distant tumor-bearing volume [(611.83 ± 113.02) mm3] was decreased than model group and CpG group [(1 406.62 ± 51.09) mm(3), (1 380.01 ± 51.44) mm3] (F = 247.160, P < 0.05), but there was no significant difference between the CpG group and the model group (P > 0.05). Serum IL-12 concentration (23.90 ± 2.33 pg/ml), IFN-γ concentration (103.02 ± 6.10 pg/ml) and spleen CD8(+)T cell ratio (4.54 ± 0.62%) in the model group were lower than those in the normal group (P < 0.05). Serum IL-12 concentration in CpG group and CpG+4-1BB group (29.21 ± 2.23 pg/ml, 37.04 ± 1.49 pg/ml), IFN-γ concentration (116.12 ± 4.08 pg/ml, 138.65 ± 1.72 pg/ml), CD8(+)T cell ratio (6.65 ± 0.64%, 12.73 ± 0.88%) were higher than the model group, while CpG+4-1BB group was higher than the CpG group (P < 0.05). The survival rate of CpG+4-1BB group was higher than that of model group and CpG group (χ(2) = 25.544, P < 0.05), but there was no significant difference between CpG group and model group (P > 0.05). There was no significant difference in organ index between the four groups (P > 0.05). Conclusion: 4-1BB monoclonal antibody combined with CpG-ODN therapy can shrink hepatoma-bearing capacity, inhibit the growth of distant tumors and significantly prolong the survival time of mice.

MiR-421 expression independently predicts unfavorable overall survival in patients with esophageal adenocarcinoma.

OBJECTIVE: This study aimed to examine the prognostic value of miR-421 in terms of overall survival (OS) and recurrence free survival (RFS) in ESAD and its potential regulatory network. PATIENTS AND METHODS: An in-silico analysis was conducted using data from large databases, including The Cancer Genome Atlas-Esophageal Carcinoma (TCGA-ESCA), Starbase 3.0 and GeneMANIA. RESULTS: Both esophageal adenocarcinoma (ESAD) and esophageal squamous cell carcinoma (ESCC) tissues had significantly upregulate miR-421 expression, compared with adjacent normal tissues. Upregulated miR-421 expression was associated with shorter OS, but not RFS in ESAD. In patients with ESCC, no difference in miR-421 expression was observed regards to OS or RFS status. Univariate and multivariate analysis showed that high miR-421 expression was independently associated with shorter OS (HR: 2.77, 95%CI: 1.41-5.46, p<0.01), after adjustment of histological grade and pathological stages. The predicted regulatory network of miR-421 in ESAD includes both tumor suppressors and oncogenes, which makes the role of miR-421 quite mysterious in this cancer. CONCLUSIONS: MiR-421 expression might serve as a valuable prognostic biomarker in patients with ESAD. But future molecular studies are required to explore the exact regulatory effect of it.

LncRNA FER1L4 suppressed cancer cell growth and invasion in esophageal squamous cell carcinoma.

OBJECTIVE: To investigate the regulatory effect of long non-coding ribonucleic acid (lncRNA) FER1L4 on biological behaviors of esophageal squamous cell carcinoma (ESCC) cells, such as proliferation and invasion. PATIENTS AND METHODS: The expressions of FER1L4 were detected in 42 pairs of ESCC tissues and corresponding para-carcinoma tissues and 5 kinds of ESCC cell lines via quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Polyethyleneimine (PEI) and liposomes were used for FER1L4 expression or interference elimination assays, respectively. The proliferation and invasion of ESCC cells were detected via MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide), apoptosis assay, cell cycle assay, and transwell chamber. RESULTS: Results of qRT-PCR showed that, compared with that in normal tissues, FER1L4 was lowly expressed in ESCC tissues. Overexpression of FER1L4 could inhibit cell proliferation and invasion, promote apoptosis and increase the cell cycle distribution in G0/G1 phase. Knockout of FER1L4 could promote the proliferation and invasion of ESCC cells, inhibit apoptosis and decrease the cell cycle distribution in G0/G1 phase. CONCLUSIONS: FER1L4 is involved in the occurrence and development of ESCC and plays a key role as a tumor suppressor gene in ESCC.

[Clinical application value of prognostic nutritional index for predicting survival in patients with advanced non-small cell lung cancer].

Objective: To explore the clinical application value of prognostic nutritional index(PNI) for predicting overall survival(OS) in patients with advanced non-small cell lung cancer (NSCLC). Methods: 123 patients with histologically confirmed non-small cell lung cancer were enrolled in this study, and their clinical and laboratory data were reviewed. The PNI was calculated as 10×serum albumin value+ 5×total lymphocyte countin peripheral blood.Univariate and multivariate analyses were used to identify the potential prognostic factors for advanced NSCLC. Results: PNI of the 123 NSCLC patients was 46.24±6.56. PNI was significantly associated with age, weight loss and pleural effusion (P<0.05). However, it showed no relationship with sex, smoking, hemoptysis, chest pain, dyspnea, histological type, clinical stage, and administration of chemotherapy (P>0.05). The median OS of the 123 patients was 19.5 months. The median OS in the higher PNI group (PNI≥46.24) and lower PNI group(PNI<46.24) were 25.2 months and 16.4 months, respectively.The 1-year survival rates were 80.6% and 63.9%, and 2-year survival rates were 54.8% and 19.6%, respectively (P<0.01). Univariate analysis showed that PNI, age, dyspnea, and weight loss were related to the OS of the advanced NSCLC patients (P<0.05). Multivariate analysis identified PNI as an independent prognostic factor for OS of advanced NSCLC (P<0.001). Conclusion: PNI can be easily calculated, and may be used as a relatively new prognostic indicator for advanced NSCLC in clinical practice.

A new angle and its relationship with early fixation failure of femoral neck fractures treated with three cannulated compression screws.

BACKGROUND: The Pauwels angle has been used widely, however an accurate evaluation of this angle is difficult because of deformity of the affected lower extremity. Therefore we designed a new measurement of the orientation of femoral neck fracture and applied this in a retrospective study to assess: (1) its reproducibility, (2) its advantages compared with the Pauwels angle, (3) its relationship with the short-term prognosis treated with three cannulated compression screws. HYPOTHESIS: This new measurement is reproducible and has some reference meaning for the treatment of femoral neck fractures. METHODS: Two hundred and twenty-eight patients with femoral neck fractures treated with three cannulated compression screws were retrospectively analyzed. The VN angle, which was the angle between the fracture line and the vertical of the neck axis, and the Pauwels angle were measured respectively. The method of ICC was performed to assess the reproducibility of the two angles, and the absolute value of difference in pre-operative and post-operative radiographs was used to evaluate the uniformity of the two angles. These fractures were divided into four groups according to VN angle (VN<0° (n=92), 0°≤VN<10° (n=82), 10°≤VN<15° (n=26), VN≥15° (n=28)), and the short-term (within 6 months) fixation results of radiographs in these fractures were evaluated. RESULTS: The ICC of the VN angle and the Pauwels angle in pre-operative radiographs were 0.937 (95% confidence interval (CI): 0.922-0.950) and 0.942 respectively (95% CI: 0.914-0.970), indicating both angles had a good inter-rater reproducibility. However, there was a great difference between the Pauwels angle in pre-operative and post-operative radiographs (P=0.037), the absolute difference was 10.66±6.47 (range: 1.72-38.48), while no statistical difference for the VN angle (P=0.084) and the absolute difference was 2.20±1.63 (range: 0.05-7.56). The overall fixation failure rate which was defined as screw loosening, varus collapse, obvious fracture displacement or femoral neck shortening was 11.84%, and the mean failure rates according to VN angles were respectively 0%, 3.24% (95% CI: 1.64-4.84), 22.69% (95% CI: 16.43-28.96), 65.45% (95% CI: 59.36-71.53). The mean failure rates of fractures according to post-operative Pauwels angle (<30°, 30-50°, >50°) were respectively 0%, 1.46% (95% CI: 1.42-1.50) and 36.24% (95% CI: 34.93-37.54). DISCUSSION: The VN angle has a good inter-rater reproducibility, a higher reliability than the Pauwels angle and is closely related to the short-term prognosis of femoral neck fractures treated with cannulated compression screws. LEVEL OF EVIDENCE: Level IV, retrospective diagnostic study.

[Association between hematocrit and risk of incident hypertension: a cohort study].

Objective: To explore the association between hematocrit level and risk of incident hypertension. Method: Subjects who participated at least two times routine health check-up in Health Management Center of Shandong Province Hospital between January 2005 and January 2010 were eligible for inclusion. After excluding participants with known hypertension and other related diseases, a prospective cohort with 20 606 subjects (female: 8 218, male: 12 388) was established. Cox-proportional hazard model was used to assess the association between hematocrit and the development of hypertension for female and male respectively. Results: During the 51 352 person-years of follow-up, newly developed hypertension was confirmed in 3 695 cases. For female, the age-adjusted hazard ratios (95%CI) for incident hypertension were 1.00 (reference), 1.06(0.86-1.29), 1.37(1.14-1.65), 1.60(1.34-1.92), respectively (P for trend<0.000 1) through the 3 quartiles of hematocrit levels.After adjusting multiple factors (age, smoking, drinking habit, physical activity, body mass index(BMI), systolic blood pressure, fasting blood glucose, serum creatinine, high-density lipoprotein, gamma-glutamyl transferase, white blood count), the hazard ratios (95%CI) were 1.00(reference), 1.05(0.85-1.29), 1.25(1.03-1.51), and 1.22(1.00-1.48), respectively (P for trend=0.016 9). For male, the hazard ratio (the highest vs. the lowest hematocrit level) after adjusting age or age and life style factors (smoking, drinking habit, physical activity) was 1.23 (1.11-1.37), 1.21(1.09-1.35), respectively. Other analyses of relationship hematocrit with incident hypertension were not statistically significant in male. Conclusions: Higher hematocrit level is associated with higher risk of incident hypertension, especially in female.

Correlation between vascular endothelial growth factor and quantitative dual-energy spectral CT in non-small-cell lung cancer.

AIM: To explore the correlation between the expression level of vascular endothelial growth factor (VEGF) in non-small-cell lung cancer (NSCLC) tissue and quantitative parameters of spectral computed tomography (CT) imaging. MATERIALS AND METHODS: Forty-eight patients with NSCLC underwent a spectral CT imaging protocol before surgical tumour resection. The repetition of iodine concentration, water concentration, and CT values at 40 keV in the region of interest were measured. The slopes of spectral attenuation curves (λHU) in the region were also calculated. The level of VEGF expression in the tumour tissue was measured using an immunohistochemical method (MaxVision method). The quantitative parameters of spectral CT imaging were compared among different levels of VEGF expression using one-way analysis of variance (ANOVA) and least significant difference (SLD) test. The correlation between VEGF expression and spectral CT imaging was estimated through Spearman's rank correlation analysis. RESULTS: There were significant differences in iodine concentrations, λHU, and CT values at 40 KeV in NSCLC between the groups showing negative and moderately positive expression of VEGF (p=0.001, 0.000, and 0.001, respectively) as well as between those showing mildly and moderately positive expression of VEGF (p=0.047, 0.005, and 0.002, respectively). In addition, all of the iodine concentrations, λHU values, and CT values at 40 KeV displayed a significant and positive correlation with the level of VEGF expression (r=0.413, 0.458, and 0.393, respectively, p<0.05). CONCLUSIONS: Quantitative parameters of spectral CT imaging may be helpful for evaluating the status of angiogenesis in NSCLC.

[Clinical characteristics and prognosis in 12 patients with adult T cell leukemia/lymphoma confirmed by HTLV-1 provirus gene detection].

Objective: To analyze the clinical characteristics and prognosis of adult T cell leukemia/lymphoma (ATLL). Methods: Peripheral blood samples from patients who were suspected as ATLL from March, 2013 to July, 2015, were collected for HTLV-1 provirus genes detection in genomic DNA extraction by PCR. Cases showing positive results were confirmed as ATLL. Clinical and laboratory characteristics, therapeutic outcomes and survival evaluation were collected. Results: 12 out of 23 suspected patients were confirmedly diagnosed as ATLL through HTLV-1 provirus genes detection by PCR. Eight patients were male and four patients were female. Median age was 51 (range 28-66) years old. All of those patients came from coastal cities of Fujian province where a HTLV-1 epidemic area locates. In the subtype classification of these 12 ATLL, 11 patients were classified as acute type and one case as lymphoma type ATLL. As one of the clinical characteristics of ATLL, ' flower cells ', with typical or atypical morphology had been observed in a high rate (81.8%). Clinical symptom such as hepatomegaly, splenomegaly and lymphadenectasis were detected in most of patients, and hypercalcemia and elevated LDH were also noted commonly. The ATLL cells immunophenotype were typical, and the major subtype was CD4+ CD8- type. Confection of hepatitis B virus was detected in a high rate (54.5%). Ten patients received chemotherapy, and 2 cases in complete remission after chemotherapy received allogeneic hematopoietic stem cell transplantation. At the end of the follow-up, 7 cases died, 4 cases survived, 1 case was lost, and the median survival was 2.8 (0.9-10.8) months. We found a case had HTLV-1 provirus negative after transplantation. Conclusion: In the coastal area of Fujian Province, ATLL is not rare. Characteristics of those ATLL are typical. But prognosis is still unsatisfactory.

Cladosporium cladosporioides Identified in China on Tobacco Seeds.

Tobacco (Nicotiana tabacum L.) is a leafy, annual, solanaceous plant grown commercially for its leaves. China is the biggest single tobacco market and accounts for more than 40% of the global tobacco consumption (1). Tobacco seed harvested in Guiyang, Guizhou Province, China, are commonly contaminated or infected by various fungal pathogens, which can cause abnormal seedlings with dark brown lesions and stunting of roots and decayed seeds. In 2013, five samples of 500 seeds from tobacco cv. Guiyan 4 were tested for germination on moistened paper on petri dishes. On average, 35% of the seeds from all five samples developed into abnormal seedlings or were decayed and were plated onto potato dextrose agar media and grown for 5 days at 25°C in darkness to confirm the presence of a pathogen. However, one fungus was isolated from an average of 10% of the 500 seeds sampled. It was identified morphologically as Cladosporium cladosporioides (Fresen.) de Vries based on the velvety olive-brown with almost black reverse colony color and dimensions and color of conidia and conidiophores. Conidia formed in long branched chains that readily disarticulate, single celled, elliptical to limoniform, 2 to 8 (avg. 4.3) × 2 to 3 (avg. 2.1) µm. Conidia were pale to olive brown and smooth to verruculose. Ramoconidia were 0 to 1 septate, 7 to 14 (avg. 9.2) × 2 to 4 (avg. 2.6) µm, smooth or sometimes minutely verruculose. Conidiophores were pale to olive brown, macro- and micronemateus, smooth or sometimes verruculose, and of various lengths up to 320 µm long and 2 to 5 µm wide. Primer pair ITS1 and ITS4 was employed to amplify the regions of ITS1-5.8s-ITS2 of the pathogens. Sequences of all three isolates (G3, G10, and G18) (Accession Nos. KF841547, KF841554, and KF841560) were identical to each other and to four sequences in GenBank (JX230994.1, JQ768317.1, JQ768322.1, and AB763555.1). Pathogenicity of the three isolates of C. cladosporioides was verified on tobacco seedlings of 3-week-old grown on wet filter paper in the petri dishes (9 cm in diameter). For each isolate, 20 seedlings incubated in one plate were inoculated with 0.5 ml of a suspension of 105 conidia/ml. Twenty seedlings were treated with sterile water as control treatment. After inoculation, the petri dishes were incubated at 25°C, 100 to 120 µEm-2 S-1, RH > 80%, and 16 h light per day for disease development. At 96 h after inoculation, symptoms comprising medium brown to black lesions on the roots were clearly visible on inoculated plants but not on the control plants. All seedlings inoculated died 9 days after inoculation whereas control seedlings remained symptomless. Re-isolation attempts on PDA from roots demonstrated C. cladosporioides to be present in symptomatic seedlings but not in roots of the control plants. Moreover, the characteristics of the cultured fungi were exactly the same as those originally isolated. Isolates G3, G10, and G18 (KF841547, KF841554, and KF841560) were deposited with the Tobacco Diseased Fungi, Guizhou Academy of Tobacco Sciences, Guizhou, China. Previously, C. cladosporioides has also been isolated from macadamia (Macadamia integrifolia Maiden & Betche) racemes in South Africa (4), from diseased papaya (Carica papaya L.) in Taiwan province of China (2), and from seeds of Amaranthus spp. in Poland (3). To the best of our knowledge, this is the first report of C. cladosporioides causing seed disease on tobacco in China and the disease should be considered in existing disease management practices. References: (1) British American Tobacco Annual Report, 8, 2012. (2) R. S. Chen, et al. Plant Dis. 93:426, 2009. (3) W. Pusz. Phytopathologia 54:15, 2009. (4) N. van den Berg et al. Plant Dis. 92:484, 2008.

Suppression of outward K⁺ currents by WIN55212-2 in rat retinal ganglion cells is independent of CB1/CB2 receptors.

Cannabinoid CB1 receptor (CB1R) signaling system is extensively distributed in the vertebrate retina. Activation of CB1Rs regulates a variety of functions of retinal neurons through modulating different ion channels. In the present work we studied effects of this receptor signaling on K(+) channels in retinal ganglion cells by patch-clamp techniques. The CB1R agonist WIN55212-2 (WIN) suppressed outward K(+) currents in acutely isolated rat retinal ganglion cells in a dose-dependent manner, with an IC50 of 4.7 µM. We further showed that WIN mainly suppressed the tetraethylammonium (TEA)-sensitive K(+) current component. While CB1Rs were expressed in rat retinal ganglion cells, the WIN effect on K(+) currents was not blocked by either AM251/SR141716, specific CB1R antagonists, or AM630, a selective CB2R antagonist. Consistently, cAMP-protein kinase A (PKA) and mitogen-activated protein kinase (MAPK)/extracellular signal-regulated kinase (ERK) signaling pathways were unlikely involved in the WIN-induced suppression of the K(+) currents because both PKA inhibitors H-89/Rp-cAMP and MAPK/ERK1/2 inhibitor U0126 failed to block the WIN effects. WIN-induced suppression of the K(+) currents was not observed when WIN was intracellularly applied. Furthermore, an endogenous ligand of the cannabinoid receptor anandamide, the specific CB1R agonist ACEA and the selective CB2R agonist CB65 also suppressed the K(+) currents, and the effects were not blocked by AM251/SR141716 or AM630 respectively. All these results suggest that the WIN-induced suppression of the outward K(+) currents in rat retinal ganglion cells, thereby regulating the cell excitability, were not through CB1R/CB2R signaling pathways.

Free vascularised fibular graft for post-traumatic osteonecrosis of the femoral head in teenage patients.

Free vascularised fibular grafting has been reported to be successful for adult patients with osteonecrosis of the femoral head (ONFH). However, its benefit in teenage patients with post-traumatic ONFH has not been determined. We evaluated the effectiveness of free vascularised fibular grafting in the treatment of this condition in children and adolescents. We retrospectively analysed 28 hips in 28 patients in whom an osteonecrotic femoral head had been treated with free vascularised fibular grafting between 2002 and 2008. Their mean age was 16.3 years (13 to 19). The stage of the disease at time of surgery, and results of treatment including pre- and post-operative Harris hip scores, were studied. We defined clinical failure as conversion to total hip replacement. All patients were followed up for a mean of four years (2 to 7). The mean Harris hip score improved from 60.4 (37 to 84) pre-operatively to 94.2 (87 to 100) at final follow-up. At the latest follow-up we found improved or unchanged radiographs in all four initially stage II hips and in 23 of 24 stage III or IV hips. Only one hip (stage V) deteriorated. No patient underwent total hip replacement. Free vascularised fibular grafting is indicated for the treatment of post-traumatic ONFH in teenage patients.

Changes in femoral head blood supply and vascular endothelial growth factor in rabbits with steroid-induced osteonecrosis.

The molecular aetiology of steroid-induced osteonecrosis (ON) is unclear. The aim of this study was to investigate changes in the femoral head blood supply and vascular endothelial growth factor (VEGF) protein levels following steroid-induced ON of rabbit femoral heads in the early stage of the disease, and to investigate a possible mechanism for ON. Using a classic protocol, ON was induced in 30 male 28-week old New Zealand white rabbits. An additional 15 untreated rabbits served as controls. Change of blood supply in the proximal femur was assessed by dynamic magnetic resonance imaging and microangiography. The VEGF protein and mRNA levels were assessed by immunohistochemistry and quantitative real-time polymerase chain reaction, respectively. After 6 weeks, the results indicated that VEGF protein and mRNA levels were significantly lower and femoral head blood supply had also decreased significantly in ON(+) rabbits compared with controls. The down-regulation of VEGF may play a critical role in the disease process of ON.

Reverse LISS plating for intertrochanteric hip fractures in elderly patients.

BACKGROUND: Fractures of the intertrochanteric hip are common and the treatment of unstable fractures generally requires an operative approach. In elderly patients, osteoporosis makes internal fixation problematic and frequently contributes to failed fixation and poor clinical results. We have attempted to apply the Less Invasive Stabilization System (LISS) in reverse position for the repair of intertrochanteric hip fractures in elderly patients with osteoporotic bones. A retrospective review is presented of the cases of 28 elderly patients with stable and unstable fractures of the intertrochanteric hip treated using the reverse LISS. METHODS: We treated 28 elderly patients with a mean age of 82.3 years. According to the Evens classification, there were 2 Type I fractures, 2 Type II fractures, 3 Type III fractures, 13 Type IV fractures, 6 Type V fractures and 2 Type R fractures. All fractures were treated using the reverse LISS. Radiographic and clinical evidence of functional outcome and complications were evaluated. RESULTS: Mean perioperative blood loss was 92.4 milliliters (range 35 to 245 milliliters), and the mean postoperative hospital stay was 8.7 days (range 3 to 14 days).Complications included one minor wound hematoma. Radiographically, no collapses, screw cutouts, or head penetrations were seen. All surviving patients (28 of 28; 100 percent) had uneventful fracture healing with union achieved by six months in all patients. CONCLUSIONS: Use of the Reverse LISS plating for intertrochanteric hip fractures resulted in event-free fracture healing.

The prophylactic effects of platelet-leucocyte gel in osteomyelitis: an experimental study in a rabbit model.

Platelet-leucocyte gel (PLG), a new biotechnological blood product, has hitherto been used primarily to treat chronic ulcers and to promote soft-tissue and bone regeneration in a wide range of medical fields. In this study, the antimicrobial efficacy of PLG against Staphylococcus aureus (ATCC 25923) was investigated in a rabbit model of osteomyelitis. Autologous PLG was injected into the tibial canal after inoculation with Staph. aureus. The prophylactic efficacy of PLG was evaluated by microbiological, radiological and histological examination. Animal groups included a treatment group that received systemic cefazolin and a control group that received no treatment. Treatment with PLG or cefazolin significantly reduced radiological and histological severity scores compared to the control group. This result was confirmed by a significant reduction in the infection rate and the number of viable bacteria. Although not comparable to cefazolin, PLG exhibited antimicrobial efficacy in vivo and therefore represents a novel strategy to prevent bone infection in humans.

Rat Dlx5 is expressed in the subventricular zone and promotes neuronal differentiation

The molecular mechanisms and potential clinical applications of neural precursor cells have recently been the subject of intensive study. Dlx5, a homeobox transcription factor related to the distal-less gene in Drosophila, was shown to play an important role during forebrain development. The subventricular zone (SVZ) in the adult brain harbors the largest abundance of neural precursors. The anterior SVZ (SVZa) contains the most representative neural precursors in the SVZ. Further research is necessary to elucidate how Dlx5-related genes regulate the differentiation of SVZa neural precursors. Here, we employed immunohistochemistry and molecular biology techniques to study the expression of Dlx5 and related homeobox genes Er81 and Islet1 in neonatal rat brain and in in vitro cultured SVZa neural precursors. Our results show that Dlx5 and Er81 are also highly expressed in the SVZa, rostral migratory stream, and olfactory bulb. Islet1 is only expressed in the striatum. In cultured SVZa neural precursors, Dlx5 mRNA expression gradually decreased with subsequent cell passages and was completely lost by passage four. We also transfected a Dlx5 recombinant plasmid and found that Dlx5 overexpression promoted neuronal differentiation of in vitro cultured SVZa neural precursors. Taken together, our data suggest that Dlx5 plays an important role during neuronal differentiation.

Curative effect and safety of vascularized fibula grafting in renal transplant recipients with osteonecrosis of the femoral head: three case reports.

Osteonecrosis of the femoral head is a common and severe complication after renal transplantation. It is characterized by deterioration of hip joint function, which impairs quality of life. We present 3 renal transplant case reports of patients with osteonecrosis of the femoral head who underwent free vascularized fibular grafting at our hospital. Follow-up was from 1(1/2) to 2 years. All 3 patients exhibited good recovery with substantial improvement in joint function. Intraoperative and postoperative findings demonstrated the safety of this surgical procedure.

The isoflavonoid daidzein attenuates the oxidative damage induced by polychlorinated biphenyls on cultured chicken testicular cells.

The soy isoflavonoid daidzein (DAI) is one of the most abundant phenolic compounds in the human diet and in animal feedstuffs. Daidzein possesses a wide spectrum of physiological and pharmacological functions related to human health. The aim of this study was to evaluate the effect of DAI on oxidative damage induced by a potential carcinogen, polychlorinated biphenyl. Testicular cells were dispersed from 18-d-old chicken embryos and exposed to DAI alone or in combination with a mixture of polychlorinated biphenyls, Aroclor 1254 (A1254), in culture. Oxidative damage was estimated by measuring the lipid peroxidation and superoxide dismutase activities and glutathione content. Results showed that DAI (10 microg/mL) increased germ cell numbers, which were inhibited by cotreatment with the estrogen receptor antagonist tamoxifen at 0.1 microg/mL. Exposure to A1254 (10 microg/mL) reduced superoxide dismutase activity and glutathione content but increased lipid peroxidation significantly. However, simultaneous supplementation with 10 microg/mL of DAI restored these parameters. The above results indicated that DAI may exert weak estrogenic activity, and more important, that DAI may display an antioxidant effect to prevent oxidative damage induced by the oxidant A1254.

Increased expression of nucleophosmin/B23 in hepatocellular carcinoma and correlation with clinicopathological parameters.

Nucleophosmin (NPM, B23, numatrin, NO38) is an abundant nucleolar phosphoprotein involved in multiple cellular functions. Previous evidence indicates that high-level expression of NPM causes uncontrolled cell growth and suggests that NPM may have oncogenic potential. In this study, we examined NPM expression in 103 paired cases of hepatocellular carcinoma (HCC), 12 cases of hepatic focal nodular hyperplasia, 17 cases of liver tissue adjacent to a hepatic haemangioma, and series of array tissues from normal human organs and malignancies using a monoclonal antibody against NPM and reverse transcription-PCR techniques, Western blot analysis, immunohistochemistry, and immunocytofluorescence. Our data indicated that NPM expression was significantly higher in HCC than in the non-malignant hepatocytes (P<0.001). Nucleophosmin was weakly expressed in hepatocytes from a 5-month-old embryo and in stationary hepatocytes of healthy adults. Moreover, enhanced expression of NPM in HCC correlated with the level of PCNA (R(2)=0.5639) and with the clinical prognostic parameters such as serum alpha fetal protein level, tumour pathological grading, and liver cirrhosis (P<0.05). Our results suggest that NPM may play an important role in the progression of tumorigenesis and that NPM may serve as a potential marker for HCC.