AIM: To explore whether strontium ranelate ( Sr ) promotes osteogenic differentiation of rat bone mesenchymal stem cells (BMSCs) through the Hedgehog/Gli1 signaling pathway.METHODS:BMSCs were isolated from 4-week-old rats by adherent culture and induced to differentiate into osteoblasts .According to the experimental purposes , the cells were exposed to different concentrations of Sr , cyclopamine ( Cy, an inhibitor of Hedgehog receptor ) or Gli1-siR-NA.The expression of Gli1 and Runx2 in the cells was detected by Western blotting .The activity of alkaline phosphatase ( ALP) was measured by the method of colorimetry , and the mineralized nodules were observed under microscope with aliz-arin red staining .RESULTS:Exposure to Sr at concentrations of 0.1 to 5 mmol/L for 7 d markedly increased the expres-sion of Gli1 in the BMSCs , and the increase in Gli1 expression was the most obvious following Sr exposure at concentration of 3 mmol/L.Cy at concentration of 10 μmol/L inhibited Sr-induced up-regulation of Gli1 expression.Transfection of the BMSCs with Gli1-siRNA not only obviously inhibited Sr-induced up-regulation of Gli1 and Runx2 ( a downstream protein of Gli1) expression, but also antagonized Sr-induced enhancement of ALP activity and the formation of mineralized nodules . CONCLUSION:The Hedgehog/Gli1 pathway is involved in Sr-induced osteogenic differentiation of rat BMSCs .

AIM: To investigate whether Janus kinase/signal transducer and activator of transcription ( JAK/STAT) signaling pathway mediates high glucose-induced damage in human umbilical vein endothelial cells ( HUVECs ) . METHODS:The cell viability was examined by CCK-8 assay.The expression levels of JAK2, STAT3, caspase-9 and en-dothelial nitric oxide synthase ( eNOS) were detected by Western blot .The intracellular levels of reactive oxygen species ( ROS) were tested by DCFH-DA staining followed by photofluorography .Mitochondrial membrane potential ( MMP) was measured by rhodamine 123 staining followed by photofluorography .RESULTS:Treatment of HUVECs with high glucose (40 mmol/L glucose) for 6~12 h enhanced the protein level of phosphorylated JAK 2, peaking at 9 h.Treatment of the cells with high glucose for 6~12 h also increased the protein level of p-STAT3 with the peak value at 12 h.Pretreatment with the inhibitor of JAK/STAT pathway AG490 for 1 h before exposure of the HUVECs to high glucose significantly inhibi-ted the high glucose -induced injury , as evidenced by an increase in the cell viability , decreases in the expression of caspase-9 and the intracellular ROS production , and increases in MMP and the expression of eNOS .CONCLUSION:JAK/STAT signaling pathway is involved in the high glucose-induced damage in HUVECs .

A comprehensive HPLC-DAD-MS method was developed to study the chemical components of semi-bionic extract of Coptis-Evodia herb couple. The extract was isolated on a Hypersil BDS C18 column (4.6 mm x 200 mm, 5 microm) using acetonitrile-ammonium formic buffer as mobile phase by gradient elution. Detection was performed on DAD and MS equipped with an electrospray ionization (ESI) source by full scan and product full scan on positive mode. The chromatogram of Coptis-Evodia showed seventeen main peaks, eight of which were from Evodia while the others were from Coptis. By comparison of the retention time, the on-line UV spectra and MS spectra, four peaks were identified as jatrorrhizine, hydroxevodiamine, palmatine and berberine, and three peaks were deduced as epiberberine, columbamine and coptisine. In addition, berberine and palmatine were quantitatively determined. No new component was created in the semi-bionic extract of the herb couple, yet the solubilities of berberine and palmatine decreased.
Berberine ; analogs & derivatives ; chemistry ; isolation & purification ; Berberine Alkaloids ; chemistry ; isolation & purification ; Chromatography, High Pressure Liquid ; methods ; Coptis ; chemistry ; Drugs, Chinese Herbal ; Evodia ; chemistry ; Plant Extracts ; chemistry ; Spectrometry, Mass, Electrospray Ionization

OBJECTIVETo explore CT and MRI manifestations of the axial area peripheral primitive neuroectodermal tumors (pPNETs) in order to improve the knowledge of this disease.

METHODSThe clinical data of 10 patients with pPNETs underwent pathologically confirmed were retrospectively analyzed from October 2008 to May 2014. There were 7 males and 3 females, aged from 8 to 49 years old with median of 23.6 years. The preoperative multi-slice spiral CT scan was completed in 3 cases, plain CT scan and enhancement in 4 cases; MRI and enhancement scanning in 5 cases; and among them, 2 cases underwent both MRI and CT scan.

RESULTSIn-bone type was found 6 cases and out-bone type was found 4 cases. Three cases occurred in sacral vertebrae, 2 cases in lumbar vertebrae, 1 case in cervical vertebrae, 1 case in cervical spinal canal, 1 case in coccyx, 1 case in the right iliac bone, 1 case in presacral space. Cross sectional the smallest tumor maximum level was 1.1 cmx 1.2 cm in size, the biggest tumor was 8.0 cm x 9.2 cm, the median size was 4.4 cm x 5.7 cm, of them, the tumor of maximal diameter larger than 5 cm had 6 cases. Except 2 cases-without destruction of bone, the other 5 cases with osteolytic destruction, 2 cases with calcification, 1 case with mixed. Equidensite was main in CT scan, 1 case with uniform density, other 6 cases with uneven density,in which 3 cases with "floating ice" change; 1 case with moderate strengthening, other 3 cases with obviously strengthening, 2 cases with multiple small blood vessels in enhancement scanning. MRI of 5 cases showed the signal of isointensity on T1WI, the slightly high signal on T2WI and the signal was not uniform; after enhancement scan, the signal of 5 cases obviously enhanced. Two patients complicated with vertebral compression fractures, no periosteal reaction was found in all patients, and no the destruction of intervertebral disk was found in 5 patients of MRI scan.

CONCLUSIONThe axial area pPNETs is common among children and the youth, and the mass often is huge. The mass of in-bone type often envelopes the vertebral body, and main located on prevertebral space, all associated with bone destruction, osteolytic destruction is common, and primary vertebral bodies also is common, attachment primary or involvement is few found, it can involve the spinal canal and anterior wall of spinal canal is common, some cases complicate with multiple newly born small vessels. The mass of out-hone type in deep soft tissue is common, minority primary spinal canal, many complicated with vertebral bone destruction, osteolytic destruction was main. The intervertebral disk was not invaded and intervertebral space has not stenosis. CT scan offer complicate with "floating ice" sign, and in-bone type is common. Isointensity is main on MRI TlWI and slightly longer signal is main on MRI T2WI, strengthening signal is obvious.

Adolescent ; Adult ; Child ; Cross-Sectional Studies ; Diagnosis, Differential ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Neuroectodermal Tumors, Primitive, Peripheral ; diagnosis ; diagnostic imaging ; Radionuclide Imaging ; Tomography, X-Ray Computed ; Young Adult

OBJECTIVETo study the diagnostic value of diffusion tensor imaging (DTI) in cervical spondylotic myelopathy.

METHODSTwenty healthy volunteers and fifty patients with cervical spondylotic myelopathy underwent DTI in the Affiliated Hospital of Medical College of Ningbo University from January 2014 to April 2015. Healthy volunteers served as controls. Fifty patients were divided into three groups (group A , B, C) according to cervical MRI scan standard. Group A (17 cases) had only the dura mater spinalis compressed; Group B (23 cases) showed the cervical spinal cord compressed, but no high signal in it; Group C (10 cases) had the cervical spinal cord compressed with high signal in the same level. The average apparent diffusion coefficients(ADC) and fractional anisotropy (FA)values in these examinee were analyzed and all subjects were performed fiber tracking.

RESULTSThere was no statistically significant differences in ADC and FA values in C2/C3, C3/C4, C4/C5, C5/C6, C6/C7 of control group (P>0.05). The average ADC and FA values in control group were (0.875 +/- 0.096) x10(3) mm2/s and 0.720 +/- 0.051, respectively; compared with group A,there was no statistically significant difference; compared with group B and C, there was significant difference; comparison among group A, B, C, there was significant differences.

CONCLUSIONDTI can early and accurately quantify the changes of microstructure in cervical spondylotic myelopathy. Fiber tracking can show the damage range of spinal cord lesions.

Adult ; Case-Control Studies ; Cervical Vertebrae ; diagnostic imaging ; Diffusion Tensor Imaging ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Radiography ; Spinal Cord Diseases ; diagnostic imaging ; surgery ; Spondylosis ; diagnostic imaging ; Young Adult

BACKGROUND: There is growing evidence that 5-fluorouracil (5-FU) combined with therapeutic trauma can effectively induce skin repigmentation in vitiligo patients who are unresponsive to conventional treatments. Previous studies have mainly focused on identifying the antimitotic activity of 5-FU for the treatment of skin cancer, but few studies have investigated its extra-genotoxic actions favoring melanocyte recruitment. METHODS: We utilized the full thickness excisional skin wound model in Dct-LacZ transgenic mice to dynamically assess the migration of melanocytes in the margins of wounds treated with or without 5-FU. The in-situ expression of CXCL12 was examined in the wound beds using immunofluorescence staining. Quantitative real-time polymerase chain reaction and Western blotting analyses were performed to detect the expression levels of CXCL12 mRNA and protein in primary mouse dermal fibroblasts treated with or without 5-FU. Transwell assays and fluorescein isothiocyanate (FITC)-phalloidin staining were used to observe cell migration and filamentous actin (F-actin) changes of melan-a murine melanocytes. RESULTS: Whole mount and cryosection X-gal staining showed that the cell numbers of LacZ-positive melanocytes were much higher in the margins of dorsal and tail skin wounds treated with 5-FU compared with the controls. Meanwhile, CXCL12 immunostaining was significantly increased in the dermal compartment of wounds treated with 5-FU (control vs. 5-FU, 22.47 ± 8.85 vs. 44.69 ± 5.97, P < 0.05). Moreover, 5-FU significantly upregulated the expression levels of CXCL12 mRNA (control vs. 5-FU, 1.00 ± 0.08 vs. 1.54 ± 0.06, P < 0.05) and protein (control vs. 5-FU, 1.00 ± 0.06 vs. 2.93 ± 0.10, P < 0.05) in cultured fibroblasts. Inhibition of the CXCL12/CXCR4 axis suppressed melanocyte migration in vitro using a CXCL12 small interfering RNA (siRNA) or a CXCR4 antagonist (AMD3100). CONCLUSION 5-FU possesses a pro-pigmentary activity through activation of the CXCL12/CXCR4 axis to drive the chemotactic migration of melanocytes.
Animals ; Cell Movement ; Cell Proliferation ; Chemokine CXCL12/genetics* ; Fibroblasts ; Fluorouracil/therapeutic use* ; Humans ; Mice ; RNA, Messenger ; Receptors, CXCR4

Objective　To observe the changes of the microvascular permeability after blunt chest trauma (BCT), endotoxemia and their combined injury in rats. Methods　After the establishment of the rat models of BCT, endotoxemia and their combined injury in the right lungs, the fluorescein sodium (FINa) content was measured with flurospectrophotometer in lungs 0.5, 1, 4 and 8 h after injury. Results　There was an early obvious increase of the microvascular permeability in the impact lateral (peak at half an hour after injury), and a delayed increase in the contralateral lung (peak at the 8th h) in the BCT group. The FINa content was higher in endotoxemia group than in the BCT group(P<0.05), and lower than that in the combined injury group(P<0.05) in the contralateral lung. Conclusion　Results indicate that there were different pathophysiologic processes among the 3 kinds of injury and the FINa content is a useful index to manifest the changes of microvascular permeability in tissues.

Objective To investigate the clinical outcomes of cranioplasty with polyether ether ketone(PEEK)or titanium after large craniectomy in patients.Methods Clinical data of 150 patients undergoing skull repair due to large frontotemporal skull defect in our hospital from April 2018 to June 2022 were retrospectively analyzed,and they were divided into titanium mesh group and PEEK group according to different repair materials.The conditions of surgical site infection,bleeding,subcutaneous effusion,seizure,implant rupture or exposure in the two groups were compared.Results In the PEEK group,96.3％of patients needed to implant the repair material under the temporal muscle,which was significantly higher than that in the titanium mesh group(78.1％)(P＜0.05).There were no significant differences in postoperative complications including infection,bleeding,seizure,implant rupture or leakage between the two groups(P＞0.05).However,the incidence of postoperative subcutaneous effusion in PEEK group was higher than that in titanium mesh group(14.8％VS4.2％,P＜0.05),and the difference was statistically significant.Conclusion Both titanium and PEEK can be used in cranioplasty for patients with large frontotemporal cranial defects.Subcutaneous effusion is common in patients underwent cranioplasty with PEEK postoperatively,which needs to be paid more attention.

Objective:To investigate the independent risk factors for postoperative severe pulmonary infection (SPI) in patients with severe traumatic brain injury (sTBI) and evaluate their predictive value.Methods:A retrospective cohort study was conducted to analyze the clinical data of 163 sTBI patients admitted to Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology from April 2021 and March 2023, including 101 males and 62 females, aged 20-80 years [53.0(46.0, 59.0)years]. The surgical procedures involved decompressive craniectomy, subdural hematoma removal, epidural hematoma removal, and intracranial hematoma removal. The patients were divided into SPI group ( n=62) and non-SPI group ( n=101) according to whether they had SPI postoperatively. The following data of the two groups were collected, including gender, age, preoperative Glasgow coma scale (GCS), elevated blood glucose, abnormal liver function, abnormal renal function, hemoglobin level, anemia, albumin level, hypoproteinemia, white blood cell count, neutrophil count, lymphocyte count, platelet count, neutrophil-to-lymphocyte ratio (NLR), derived neutrophil-to-lymphocyte ratio (dNLR), platelet-to-lymphocyte ratio (PLR), prognostic nutritional index (PNI) and serum lactate dehydrogenase (LDH) level. All the hematological tests were performed on venous blood samples collected preoperatively before anti-inflammatory treatment. Independent risk factors for predicting the postoperative occurrence of SPI in sTBI patients were identified through univariate analysis and multivariable stepwise regression analysis. The predictive value of separate indicator or indicators combined was assessed by calculating the area under the curve (AUC) of the receiver operating characteristic (ROC) curve. Results:Univariate analysis demonstrated that preoperative GCS, albumin level, lymphocyte count, NLR, PNI and serum LDH level in both groups were significantly correlated with the postoperative occurrence of SPI ( P<0.05), while gender, age, elevated blood glucose, abnormal liver function, abnormal renal function, hemoglobin level, anemia, hypoproteinemia, white blood cell count, neutrophil count, platelet count, dNLR and PLR were not correlated with the postoperative occurrence of SPI in sTBI patients ( P>0.05). Multivariable stepwise regression analysis revealed that low lymphocyte count (95% CI -0.337, -0.013, P<0.05), high NLR (95% CI -0.023, -0.005, P<0.01), low PNI (95% CI 0.007, 0.026, P<0.01), and high serum LDH (95% CI -0.002, -0.001, P<0.01) were independent risk factors for SPI in sTBI patients ( P<0.05). ROC curve analysis indicated that low lymphocyte count, high NLR, low PNI and high serum LDH level could predict SPI in sTBI patients postoperatively, with the combination of PNI and serum LDH showing the highest predictive ability (AUC=0.78, 95% CI 0.70, 0.85). Conclusion:Low lymphocyte count, high NLR, low PNI, and high serum LDH level are independent risk factors for postoperative SPI in patients with sTBI, and the combination of PNI and serum LDH possesses a high predictive value for postoperative SPI in sTBI patients.

OBJECTIVETo study the correlation between the expression of Fcgamma receptor II b (FcgammaRII b) on B cells and rheumatoid arthritis (RA) patients of Shen deficiency syndrome (SDS).

METHODSThere were 43 RA patients, including 26 of SDS and 17 of non-SDS. The expression levels of FcgammaRII b on naive B cells, memory B cells, and plasma blasts in the peripheral blood were detected by flow cytometry. The numbers of tender joints, numbers of swollen joints, erythrocyte sedimentation rate (ESR), rheumatoid factor (RF), and disease activity score (DAS28), the correlation between the distribution of B cells and the expression level of FcgammaRII b in RA patients were analyzed. Besides, another 21 healthy volunteers were recruited as the control group.

RESULTSThe expression level of FcgammaRII b was 49.65% +/- 15.86% on memory B cells and 43.69% +/- 22.57% on plasma blasts in RA patients of SDS, significantly down-regulated when compared with those of the control group (64.03% +/- 6.01%, 66.59% +/- 10.18%, P < 0.01). The expression level of FcgammaRII b on memory B cells of RA patients of non-SDS was down-regulated more obviously when compared with that of the control group (52.70% +/- 9.52% versus 64.03% +/- 6.01%, P < 0.01). The expression level of FcgammaRII b on plasma blasts was obviously lower in RA patients of SDS than in RA patients of non-SDS (56.10% +/- 17.05%, P < 0.05). The expression level of FcgammaRII b on memory B cells was not correlated with numbers of tender joints, numbers of swollen joints, ESR, RF, or DAS28.

CONCLUSIONSThe defective immunological tolerance of B cells in RA patients of SDS might be closely correlated with down-regulation of FcgammaRII b on memory B cells and plasma blasts. There might exist genetic abnormality of FcgammaRII b gene in RA patients of SDS, thus inducing loss of autoimmunity tolerance.

Adult ; Arthritis, Rheumatoid ; blood ; diagnosis ; immunology ; B-Lymphocytes ; immunology ; metabolism ; Case-Control Studies ; Female ; Humans ; Male ; Medicine, Chinese Traditional ; Middle Aged ; Receptors, IgG ; immunology ; metabolism