BACKGROUND: Growth differentiation factor 5 (GDF-5) is an important factor to regulate the formation and development of the cartilage and bone, it plays a crucial role on the promotion of repairing bone, cartilage and tendon ligament injury. OBJECTIVE: To transfect eukaryotic expression plasmid pcDNA 3.1(+)/GDF-5 to bone marrow stroma stem cells of mouse and to check the expression of extracellular matrix and proteoglycan which relates with the cartilage formation and differentiation. DESIGN, TIME AND SETTING: An in vitro observation regarding cells was performed in the central laboratory of Wuhan Union Hospital between March and December in 2008.MATERIALS: Twenty Kunming specimen male mice were offered by Experimental Animal Center of Tongji Medical College of Huazhong University of Science and Technology. The eukaryotic expression plasmid pcDNA 3.1(+)/GDF-5 was preserved at the laboratory.METHODS: The man-ow stroma stem cells were isolated from mouse bone marrow and cultured in vitro with whole bone marrow adherence method. Passage 3 cells were incubated on 6-well plate and began to transfect when they were 90% confluent. Experiment was assigned into three groups: trensfection group underwent transient transfection of liposome-mediated pcDNA 3.1(+)/GDF-5 using LipofectamineTM2000; blank plasmid group was transfected with blank plasmid pcDNA 3.1(+); control group was added with equal volume of liposome and other protocols were the same as above. MAIN OUTCOME MEASURES: The transfection efficacy was identified success by the expression of GDF-5 gene and protein using RT-PCR and immunocytochemistry at 72 heurs following transfection, cartilage matrix Ⅱ collagen expression was determined as above methods. Then marrow stroma stem cells were cultured for additional two weeks to check expression of proteoglycan with alcian blue staining.RESULTS: In the transfection group, a 219-bp specific amplification band was visible, there were brown positive stain in the cytoplasm of marrow stroma stem calls; In blank plasmid group and control group, no GDF-5 trensfection, specific amplification band or obvious stain of cytoplasm was observed. In the transfection group, the collagen Ⅱ gene was detected to express at 225 bp, with brown yellow stain in cytoplasm; in the blank plasmid group and control group, no collagen Ⅱ gene expression or SP ～ stain was observed. Alcian blue staining results showed the transfected cells were stained blue while those in the blank plasmid group and control group were not metachromasia stained.CONCLUSION: Gene trensfection of pcDNA 3.1 (+)/GDF-5 to marrow stroma stem cells can significantly raise expression of collagen II and proteoglycan, and promote the chondrogenic differentiation of marrow stroma stem cells.

OBJECTIVEThe aim of this study was to analyze the initial displacement of the maxillary first molar by tip back bend in Tip-Edge force system and to explore the relationships between the displacement and the degree or position of tip back bend.

METHODSThe Tip-Edge three-dimensional finite element model was constructed by CT scan of a dry skull with all teeth. The conditions of five types of degree (20 degrees, 30 degrees, 40 degrees, 50 degrees, 60 degrees) and four types of position (2, 4, 6, 8 mm) of tip back bend were added to the three-dimensional finite element model and the initial displacements of the maxillary first molar were calculated.

RESULTSUnder the effect of tip back bend, distal and extrusive displacement of the maxillary first molar occurred. The distal and extrusive movement increased as the degree of tip back bend increased or the position of tip back bend moved distally.

CONCLUSIONThe initial displacement of the maxillary first molar could be controlled by the precise control of the degree and position of tip back bend.

Finite Element Analysis ; Humans ; Maxilla ; Models, Biological ; Molar ; Periodontal Ligament

Objective To investigate the effect of melatonin against cerebral ischemia reperfusion injury (CIRI) in mice and its mechanism.Methods Thirty male C57BL/6 mice were randomly divided into sham operation group,CIRI group,and melatonin treatment group (n =10 in each group).A middle cerebral artery occlusion model was induced by suture method.The degree of brain injury was evaluated by neurological function score,brain water content,and cerebral infarction volume.Western blot analysis was used to detect apoptosis-related proteins Bim,Bcl-2,and endoplasmic reticuhm stress-related molecules C/ EBP homologous protein (C/EBP) expression.Results Compared with the CIRI group,the neurological function score was significantly improved,the degree of cerebral edema was significantly reduced,and the volume of cerebral infarction was significantly reduced in the melatonin treatment group (all P ＜0.05).In addition,the expression of Bcl-2 was significantly up-regulated in the melatonin treatment group,and the expression of Bim and CHOP was significantly down-regulated (all P ＜ 0.05).Conclusion Melatonin may play an anti-CIRI role by regulating CHOP,and endoplasmic reticulum stress plays an important role in CIRI.

Objective To observe the circadian rhythm of blood pressure and investigate the impact of Norvasc and Tanatril administrated at different time points based on the theory of time therapeutics in hypertensive patients. Methods The 24-hour ambulatory blood pressure was monitored in 130 inpatients and outpatients with grade 3 or 2 hypertension from May 2008 to November 2009. A total of 34 dipper hypertensive patients were grouped into dipper blood pressure group and 5 mg/d of Norvasc and Tanatril were taken by them at 6:00 AM. A total of 96 nondipper hypertensive patients were further divided into three subgroups after adjustment for age and gender: 5 mg/d of Norvasc and Tanatril were taken by group I (n = 30) at 6:00 AM; 5 mg/d of Tanatril at 6:00 AM and 5 mg/d of Norvasc at 18:00 PM by group Ⅱ (n = 32); 5 mg/d of Norvasc at 6:00 AM and 5 mg/d of Tanatrilat 18:00 PM by group Ⅲ(n=34).The 24-hour ambulatory blood pressure monitoring was performed again after four weeks treatment and 24-hour mean systolic blood pressure(24 hSBP),24-hour mean diastolic blood pressure(24 hDBP),daytime and nighttime mean systolic blood pressure(dSBP,nSBP)and daytime and nighttime mean diastolic blood pressure(dDBP,nDBP),were analyzed.Results The 24 hSBP,24 hDBP and dSBP,and dDBP were reduced from(154.3±5.6),(95.4±3.1),(158.7±6.5),(99.6±3.7)mmHg to(137.2±3.9),(82.5±2.7),(139.7±3.8),(85.2±3.5)mmHg,respectively,in dipper blood pressure group(t=2.124,2.356,2.278,2.449,all P<0.05).The 24 hSBP and 24 hDBP of the three groups in nondipper blood pressure were decreased from(154.4±6.1),(156.7±6.7),(156.6±5.2),(95.8±2.8),(94.9±3.8),(95.7±3.2)mmHg to (139.6±4.1),(134.5±4.6),(133.4±3.5),(83.5±4.2),(80.8±5.6),(81.6±4.7)mmHg,respectively(t=2.038,2.040,2.135,2.142,2.213,2.216,all P<0.05).dSBP and dDBP were decreased from(158.6±3.50),(158.4±5.6),(159.5±4.),(98.4±3.7),(99.6±3.7),(83.9±5.2)mmHg to(138.9±5.4),(136.7±4.1),(137.4±6.4),(85.8±5.3),(83.6±5.1),(83.9±5.2)mmHg,respectively(t=2.021,2.252,2.261,2.217,2.167,2.076,all P<0.05).nSBP and nDBP were decreased from(146.7±6.9),(149.8±3.9),(150.2±4.1),(93.7±4.2),(95.7±4.3),(93.4±3.3)mmHg to(133.7±4.6),(129.8±5.7),(127.6±2.8),(87.8±2.9),(78.5±6.4),(77.8±4.8)mmHg,respectively(t=1.798,2.032,2.014,1.864,2.157,2.166,all P<0.05).There were significant differences in nSBP and nDBP among all groups after treatment(F=2.32,2.17,all P<0.05),and the effect of the drugs was better in groug Ⅱ and Ⅲ than in group Ⅰ(q=3.17,4.03,3.32,4.19,all P<0.05),but there were no significant differences between group Ⅱ and Ⅲ(P>0.05).Conclusions Blood pressure can be controlled effectively by taking two antihypertensive medictions in the morning in hypertensive dipper patient but the blood pressure of nondipper hypertensive patients were able to be controlled more efficiently by taking the two antihypertensive medictions separately in the morning and at the evening compared with that taking the two drugs together in the morning.

The purpose of this study was to establish a model of trigeminal neuralgia (TN) through an approach from lower edge of cheekbone and to observe the functional changes in the voltage-gated potassium currents in the cultured trigeminal ganglion (TG) neurons. Thirty Sprague-Dawley male rats were divided into two groups, the sham-operated (sham) group and the operated group. The TN model was carried out by using a chronic constriction injury of the infraorbital nerve (ION-CCI) from lower edge of cheekbone. Peripheral pain threshold test and whole-cell patch clamp recording were used to determine the difference between sham and ION-CCI rats. The withdrawal threshold of whisker pad in operated side of ION-CCI rat was decreased significantly from 6 d after operation and then maintained until 21 d, with the lowest on the 15th day. The threshold of whisker pad in non-operated side of operated rats was also decreased significantly compared with that in the sham group. Delayed rectifier potassium current (I(K)) in cultured ION-CCI TG neurons was decreased significantly compared with that in the sham group. Transient outward potassium currents (I(A)) in both operated and non-operated sides of TG neurons from ION-CCI rats were also reduced significantly compared with that in the sham group. The present study provided a new method of ION-CCI. In this model, the decrease of I(A) and I(K) might contribute, at least in part, to the decrease in mechanical pain threshold of whisker pad and the subsequent hyperalgia.
Animals ; Cells, Cultured ; Constriction ; Disease Models, Animal ; Hyperalgesia ; Male ; Pain Threshold ; Patch-Clamp Techniques ; Potassium Channels ; metabolism ; Rats ; Rats, Sprague-Dawley ; Trigeminal Ganglion ; metabolism ; Trigeminal Neuralgia ; physiopathology ; Vibrissae

OBJECTIVETo evaluate the prevalence of nasal carriage of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) in healthy children from Chengdu.

METHODSStrains of Staphylococcus aureus were isolated from nasal swabs of healthy children from five kindergartens in Chengdu from September, 2005 to December, 2005 and questionnaires were obtained. Antibiotic susceptibility test was performed with agar disk diffusion and Bauer-Kirby on Mueller-Hinton medium method to determine CA-MRSA. mecA and PVL genes were detected with PCR in all of the CA-MRSA isolates.

RESULTSA total of 801 children were enrolled. Overall 147 children (18.4%) were carried with Staphylococcus aureus and 9 (1.1%) were carried with CA-MRSA. All CA-MRSA isolates were positive for mecA gene, and 5 CA-MRSA isolates were positive for PVL gene. Of the 9 CA-MRSA isolates, 6 were multiresistant.

CONCLUSIONSCA-MRSA nasal colonization is present among Chengdu healthy children. The CA-MRSA isolates are multiresistant and parts of CA-MRSA isolates carry PVL gene. The nasal carriage of CA-MRSA in healthy children should be a concerned issue.

Bacterial Proteins ; genetics ; Carrier State ; microbiology ; Child ; Child, Preschool ; China ; Community-Acquired Infections ; microbiology ; Female ; Humans ; Male ; Methicillin-Resistant Staphylococcus aureus ; genetics ; isolation & purification ; Microbial Sensitivity Tests ; Nose ; microbiology ; Penicillin-Binding Proteins

BACKGROUNDRathke's cleft cyst (RCC) is one of the most common incidentally discovered sellar lesions, while symptomatic cases are relatively rare. Surgical treatment is recommended for symptomatic patients to drain the cyst content and to remove the capsule safely. The aim of this study was to clarify the clinical features, surgery considerations and therapy outcomes of symptomatic RCCs.

METHODSTotally 42 patients (19 males and 23 females) were retrospectively reviewed with the diagnosis of RCCs under surgery resection at the Affiliated Hospital of Medical College, Qingdao University between January 2005 and December 2010.

RESULTSPatients' age ranged from 6 to 67 years (mean of 41.6 years). The duration of symptoms ranged from 4 days to 10 years. Headache (69%), visual impairment (36%), and pituitary dysfunction (10%) were the most common presenting symptoms. The maximum diameter of cysts ranged from 6.0 to 46.7 mm (mean of 20.07 mm). Of the 42 patients, 36 underwent endonasal transsphenoidal approach and the others underwent transcranial approach. Thirty patients had a subtotal resection and decompression, while 12 patients had a total cyst resection. Cysts of 28 patients were lined by simple cubical or columnar epithelium, and cysts of 34 patients were filled by amorphous colloid material, that was the characteristic of RCCs. The majority of patients presented with a simple headache, and 93% of this group experienced a complete improvement after surgery. Twelve of 15 patients (80%) with preoperative visual deficits experienced an improvement in their vision after surgery. All of those patients with pituitary dysfunction experienced an improved endocrine status. The endocrinological complication usually was diabetes insipidus, and postoperative transient diabetes insipidus occurred in 13 (31%) patients without any permanent diabetes insipidus. The overall recurrence rate was 7% at a mean follow-up of 22 months (range 12 - 60 months).

CONCLUSIONSSurgical treatment is to drain the contents of the cyst and to remove the capsule as much as possible under the precondition that does not increase the complications. Biopsy and decompression procedures are recommended for most cases.

Adolescent ; Adult ; Aged ; Central Nervous System Cysts ; diagnosis ; pathology ; surgery ; Child ; Female ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

OBJECTIVETo screen the molecular markers of DNA methylation with potential diagnostic value, and to explore their methylation features in Chinese colorectal neoplasma in order to find out ones with higher diagnostic value.

METHODSTissue samples of colorectal cancer and normal adjacent mucosa(>10 cm distance to tumors) from 10 colorectal cancer patients undergoing operation, and tissue samples of colorectal adenoma from 10 patients undergoing endoscopic resection in our center from June to August 2013 were collected respectively. Methylation status of 8 genes, such as SNCA, MAL, INA, SPG20, FBN1, CNRIP1, TFPI2, OSMR, was detected by BSP and qMSP to screen genes with potential diagnostic valua. ROC curve was drawn to analyze its diagnostic value.

RESULTSBSP measurement showed that the rate of DNA methylation of SNCA, SPG20 and FBN1 was 100% in colorectal cancer and adenoma, while no methylation was found in normal adjacent mucosa. The other 5 genes expressed in different extent in cancer, adenoma and normal adjacent mucosa. Among 10 cancer tissues and normal adjacent mucosa detected by qMSP method, positive SNCA methylation was found in 5 cases and 1 case respectively; positive SPG20 in 8 cases and 1 case respectively; positive FBN1 in 7 cases and 0 cases respectively, whose differences were significant (P=0.070, P=0.003 and P=0.007). The area under curve(AUC) of SNCA, SPG20, and FBN1 methylation for diagnosing colorectal cancer was 0.890, 0.730 and 0.880 respectively.

CONCLUSIONSNCA, SPG20 and FBN1 are potential genes with screening value for colorectal neoplasma.

We observed the effects of IKVAV self-assembling peptide nanofiber scaffold (SAPNS) on olfactory ensheathing cells (OECs). The IKVAV molecules were triggered to self-assemble to interconnected nanofibers hydrogel by adjusting pH of solution and adding of DMEM/F12 culture medium. Atomic Force Microscopy (AFM) showed that self-assembly hydrogel was consisted of the interconnected nanofibers, which varied from three nm to five nm in diameter and hundreds nanometer in length. The primary OECs were isolated from rat olfactory bulb and purified by differential adhesion twice. At days 12, the purity of OECs was 85% according to immunostaining of P75 NGFR antibody. OECs were cultured with IKVAV peptide. The adhesion, viability and proliferation of OECs were observed with inverted microscope, Calcein-AM/PI staining and Cell Counting Kit-8. OECs cultured on IKVAV SAPNS grew well and the viable cell count was 95%. IKVAV SAPNS can promote the adhesion of OECs and did no hinder the proliferation of OECs. IKVAV SAPNS nanofiber gel has good biocompatibility and bioactivity for OECs. It can serve as a good nerve tissue engineering scaffold.
Animals ; Biocompatible Materials ; chemistry ; Humans ; Hydrogels ; chemistry ; Laminin ; chemistry ; Materials Testing ; Nanofibers ; chemistry ; Olfactory Bulb ; cytology ; drug effects ; Olfactory Nerve ; cytology ; drug effects ; Peptide Fragments ; chemistry ; Rats ; Rats, Sprague-Dawley ; Tissue Engineering ; methods ; Tissue Scaffolds ; chemistry

Objective:To investigate and summarize the chest CT imaging features of patients with novel coronavirus pneumonia (COVID-19), bacterial pneumonia and other viral pneumonia.Methods:Chest CT data of 102 patients with pulmonary infection due to different etiologies were retrospectively analyzed, including 36 patients with COVID-19 admitted to Hainan Provincial People's Hospital and the Second Affiliated Hospital of Hainan Medical University from December 2019 to March 2020, 16 patients with other viral pneumonia admitted to Hainan Provincial People's Hospital from January 2018 to February 2020, and 50 patients with bacterial pneumonia admitted to Haikou Affiliated Hospital of Central South University Xiangya School of Medicine from April 2018 to May 2020. Two senior radiologists and two senior intensive care physicians were participated to evaluated the extent of lesions involvement and imaging features of the first chest CT after the onset of the disease.Results:Bilateral pulmonary lesions were more common in patients with COVID-19 and other viral pneumonia, and the incidence was significantly higher than that of bacterial pneumonia (91.6%, 75.0% vs. 26.0%, P < 0.05). Compared with other viral pneumonia and COVID-19, bacterial pneumonia was mainly characterized by single-lung and multi-lobed lesion (62.0% vs. 18.8%, 5.6%, P < 0.05), accompanied by pleural effusion and lymph node enlargement. The proportion of ground-glass opacity in the lung tissues of patients with COVID-19 was 97.2%, that of patients with other viral pneumonia was 56.2%, and that of patients with bacterial pneumonia was only 2.0% ( P < 0.05). The incidence rate of lung tissue consolidation (25.0%, 12.5%), air bronchial sign (13.9%, 6.2%) and pleural effusion (16.7%, 37.5%) in patients with COVID-19 and other viral pneumonia were significantly lower than those in patients with bacterial pneumonia (62.0%, 32.0%, 60.0%, all P < 0.05), paving stone sign (22.2%, 37.5%), fine mesh sign (38.9%, 31.2%), halo sign(11.1%, 25.0%), ground-glass opacity with interlobular septal thickening (30.6%, 37.5%), bilateral patchy pattern/rope shadow (80.6%, 50.0%) etc. were significantly higher than those of bacterial pneumonia (2.0%, 4.0%, 2.0%, 0%, 22.0%, all P < 0.05). The incidence of local patchy shadow in patients with COVID-19 was only 8.3%, significantly lower than that in patients with other viral pneumonia and bacterial pneumonia (8.3% vs. 68.8%, 50.0%, P < 0.05). There was no significant difference in the incidence of peripheral vascular shadow thickening in patients with COVID-19, other viral pneumonia and bacterial pneumonia (27.8%, 12.5%, 30.0%, P > 0.05). Conclusions:The probability of ground-glass opacity, paving stone and grid shadow in chest CT of patients with COVID-19 was significantly higher than those of bacterial pneumonia, and it was more common in the lower lungs and lateral dorsal segment. In other patients with viral pneumonia, ground-glass opacity was distributed in both upper and lower lungs. Bacterial pneumonia is usually characterized by single lung consolidation, distributed in lobules or large lobes and accompanied by pleural effusion.