Objective To investigate the coverage of a recombinant protein vaccine based on pneumococcal surface protein A (PspA) from both family 1 and family 2.Methods One hundred and fifty-nine Streptococcus pneumoniae strains, including 47 invasive strains, were isolated from children in Nanjing Children′s Hospital.Cell lysates were prepared and reacted with three antibodies recognizing PspA -RX1, PspA-3296 and PspA-5668 for PspA typing by ELISA .Results Among 47 invasive isolates of 9 different serotypes, 10.7%were PspA family 1 and 89.3%were PspA family 2.Among all of 159 clinical isolates, 10.1% were identified as PspA family 1, 88.0%were family 2, while 1.9%of strains could not be typed by ELISA and PCR assays .None of strains belonged to PspA family 3.Conclusion The recombinant pro-tein vaccine based on PspA from both family 1 and family 2 has a broad coverage among clinical isolates and is potentially protective against both invasive and non-invasive pneumococcal diseases .

UNLABELLEDOBJECTIVE To study the effects of TiO2 blasted and acid-etched surfaces of cp-titanium on changing composition of oxide-film and attachment and proliferation of osteoblasts in vitro.

METHODScp-titanium discs were prepared and divided into 4 groups: TiO2 blasted (SB), sandBlasted and acid-etching (SLA1 and SLA2) and machine-polished surface (S1). Scanning electron microscopy (SEM), X-ray diffraction (XRD) and electron probe microanalysis (EPMA) were used to test surface morphology and composition of oxide-film. Osteoblasts were cultured on the titanium surface of 4 groups. MTT assay was used to measure the attachment and proliferation.

RESULTSIn regard to surface roughness, average roughness for SB, SLA1 and SLA2 was obviously higher than S0 Oxygen ratio increased on SB,contrarily, it decreased on SLA. A mixture of anatase and rutile-type crystals were observed in the SB. Smaller anatase were observed in the SLA1 and SLA2. The oxide thickness on SLA surface was thiner than that on the SB surface. Alter 1, 4, 24 hours' culture, the number of osteoblast attachment on SB surfaces was the highest (P < 0.05). The number of cells osteoblast proliferation was the highest on SB after 1, 3, 5 and 7 days' culture (P < 0.05).

CONCLUSIONThe thickness and chemical composition of oxide film play an important role in osteoblast attachment and proliferation at the same roughness surface. It is concluded that osteobla.st attachment and proliferation are better on SB surfaces than on SLA1 and SLA2 surfaces.

Cell Adhesion ; Cell Proliferation ; Microscopy, Electron, Scanning ; Osteoblasts ; Oxides ; Surface Properties ; Titanium

Objective To analyse the characteristics of orbital benign and malignant masses on diffusion weighted imaging in combination with conventional MR imaging and evaluate the diagnostic value of apparent diffusion coefficient in distinguishing benign and malignant orbital lesions.Methods Seventy- seven cases with orbital masses,including fifty-five benign lesions and twenty-two malignant tumors,who underwent conventional MRI and diffusion imaging scanning were studied with use of a 1.5 T magnetic resonance system.Quantitative ADC measurements of masses(ADC_M)and of the white matter of eontralateral temporal lobe(ADC_W)were made with two different b-values of 0 and 1000 s/mm~2.The ADC ratio(ADCR)of the lesion to the control was calculated.The receiver operating characteristic curves(ROC) were constructed using various cut points of ADC_M and ADCR for different parameters to differentiate between benign and malignant masses.The area under the ROC curve for each parameter was also calculated. Results All cases were proved by histopathology.The mean ADC_M and ADCR of benign orbital masses were(1.56?0.75)?10~(-3)mm~2/s and 1.85?0.91,respectively.The mean ADC_M and ADCR of malignant orbital masses were(1.09?0.42)?10~(-3)mm~2/s and 1.28?0.53,respectively.There were significant difference both between ADC_M and ADCR of benign and malignant masses(t=2.803,2.735,P

UNLABELLEDOBJECTIVE To evaluate the effects of grooved, alkali- and heat-treated, acid-etched and TiO2 blasted surfaces of titanium substrates on F-actin cytoskeleton of osteoblasts in vitro.

METHODSOsteoblasts derived from fetal rat calvarial were cultured on 6 different commercially pure titanium discs-grooved(G), sandblasted (SB), sand-blasted and acid-etching (SLA) surfaces and alkali- and heat-treated (AH1, AH2, AH3) surfaces. For F-actin cytoskeleton measurement, osteoblasts whose filamentous actin was stained with phalloidin-TRITC were cultured for 1, 2, 4, 12 h, evaluated by CLSM observation.

RESULTSOsteoblasts attached to the different types of surfaces after 1 hour culture were similar. The actin cytoskeleton formed a ring of cortical filaments around the nucleus after 1 hour on SB, AH2, AH3, SLA surfaces. Actin filaments condensed along edges of pits. The actin filaments of seeded cells were spread after 2 h. The actin filaments on G formed bundles around the nucleus. The filaments began to parallel to the grooves. On AH1, the fibres formed a ring of cortical filaments around the nucleus with some cytoplasmic fibres radially oriented. On AH2, AH3, SB, the fibres orignised in a cytoplasmic meshwork with fibres which terminate at the ridge of depressions. The cell were suspending itself over the depressed areas. Actin filaments on SB were distinct and well formed that were oriented paralled to one another and the long axis of cells. After 4 h, actin filaments appeared organised in a parallel to one another and the long axis of cells. After 12 h, the actin filaments on all surfaces were well spread and were oriented paralled to another and to the long axis of the cell. The filaments formed bundles which reached to holes or adhered to the ridge of raised points, suspending cells over depressed areas.

CONCLUSIONAfter 12 h, the actin filaments on all surfaces were well spread and were oriented parallel to another and to the long axis of the cell. It was concluded that F-actin cytoskeleton of osteoblasts were spread best on SB surfaces among all surfaces.

Actin Cytoskeleton ; Actins ; Animals ; Cytoskeleton ; Microtubules ; Osteoblasts ; Prostheses and Implants ; Rats ; Surface Properties ; Titanium

OBJECTIVETo identify whether SH3GL1 gene serves as a disease associated gene of adolescent idiopathic scoliosis (AIS).

METHODSPositioning candidate cloning: "case-sibling or case-family control design" research scheme based on family constellation was designed. Fifty-six AIS patients (15 male and 41 female, mean age 15 years old, ranged from 8 to 22 years old, Cobb angle from 25 degrees to 110 degrees , average Cobb angle of 67.5 degrees ) from November 2007 to December 2008 were recruited. In all patients, blood preparation was collected, and genome DNA was extracted. According to nucleotide sequence of gene SH3GL1, primer pair for PCR amplification, cloning, and sequencing with 10 exons as emphasis was designed. Sequence comparative analysis for exon sequencing result between sib pairs or family pairs, and that between sib pair or family pairs and NCBI (National Center for Biotechnology Information) were conducted through Vector NTI Advance 10.3 software to judge whether basic group mutation occurred or not. Amino acid sequence comparative analysis for prediction was made.

RESULTSTen exons of the candidate gene SH3GL1 were successfully amplified and cloned in genome DNA of an AIS sib pair and family pairs, and the sequencing obtained positive results. Twelve basic group mutations were found in 10 exons of the candidate gene SH3GL1 of patients with AIS. These mutations were located in the second exon (3 mutations), the fourth exon (1 mutations), the fifth exon (4 mutations), the sixth exon (1 mutations), the eighth exon (1 mutations), and the tenth exon (2 mutations, noncoding region). If basic group in 515 of mRNA was mutated to T, termination codon(TAG) came into being and open reading frame was altered. The sequence of protein showed brachytmema protein was encoded, which could cause changes of primary structure.

CONCLUSIONSH3GL1 is possibly one of the disease associated genes of AIS.

Adolescent ; Base Sequence ; Child ; Exons ; genetics ; Female ; Humans ; Intracellular Signaling Peptides and Proteins ; genetics ; Male ; Scoliosis ; genetics ; Sequence Alignment ; Young Adult

AIM:To investigate the effect of ischemic postconditioning ( IPC) on autophagy induced by focal cerebral ischemia reperfusion ( I/R) in rats.METHODS:Healthy male SD rats were assigned randomly into sham-opera-tion (sham) group, I/R group and IPC group with 10 rats in each group.The rats in sham group were only exposed the right common , internal and external carotid artery surgically .The rats in I/R group were subjected to right middle cerebral artery occlusion (MCAO) by the modified Longa suture method for 2 h followed by 24 h of reperfusion.The rats in IPC group were subjected to MCAO for 2 h followed by reperfusion of the ipsilateral common carotid artery occlusion for 10 s for 5 episodes, and then reperfusion for 24 h.Autophagy was obeserved by transmission electron microscopy (TEM).The pro-tein levels of mammalian target of rapamycin (mTOR), p-mTOR and microtubule associated protein light chain 3 (LC3)-II in brain tissue of the rats were determined by Western blot .Pathological changes of brain tissue were observed by HE staining.RESULTS:The protein levels of mTOR and p-mTOR in IPC group were significantly higher than those in I/R group (P<0.05).The expression of LC3-II in IPC group was significantly lower than that in I/R group (P<0.01).The cerebral infarction area and brain water content in IPC group were significantly lower than those in I /R group (P<0.01). HE staining showed that neurons degeneration and necrosis in IPC group were significantly alleviated compared with I /R group.TEM observation showed that IPC revealed fewer autophagosomes , with much less severe cell damage than that in I/R group.CONCLUSION:IPC reduces brain ischemia reperfusion damage by decreasing autophagy of brain cells , which might be related to the activation of mTOR .

Autopsy ; Coxsackievirus Infections ; Enterovirus ; isolation & purification ; Enterovirus B, Human ; isolation & purification ; Enterovirus Infections ; Hand, Foot and Mouth Disease ; pathology ; virology ; Humans ; Infant ; Male

Objective Type 1 diabetes mice model was established to investigate the changes of key enzymes involved in testosterone synthesis in testes of early diabetic mice.Methods Tatolly 20 male C57 mice were randomly divided into two groups:control and diabetic groups,and the diabetes mice were ip administered with a single dose of 150 mg/kg Streptozotocin.Four weeks after confirmation of diabetic model,the serum and testis were collected for further study.The qRT-PCR method was used to measure the expression of LHR and steroidogenesis synthetase StAR,P450scc,3β-HSD6,P450c17a1,and 17β-HSD3 mRNA.ELISA assay was performed to measure the levels of testosterone and luteinizing hormone (LH) in serum,and the enzymatic activities of 3β-HSD1,1P450c17 and 17β-HSD3 in testis tissue.Results Compared to control group,the levels of testosterone and LH of diabetic group declined significantly (P ＜ 0.05) after four weeks.The mRNA levels of LHR,StAR,CYP11a1,Hsd3b6,CYP17a1 and Hsd17b3,and enzymatic activities of 3β-HSD6,P450c17 and 17β-HSD3 were also decreased significantly compared with control group (P ＜ 0.05,0.01 and 0.001).Conclusion Expression of key enzymes of testosterone synthesis in testis of early diabetic mice decreases significantly.

Objective　To investigate the occurrence of postural hypotension (PH) in patients suffering from type 2 diabetes mellitus with or without hypertension (DMH or DM), and the relationship of PH and diabetic neuropathy, hyperinsulinemia and insulin resistance. Methods　A total of 30 cases of type 2 DM and 30 cases of DMH were included in this study. The blood pressure of all subjects were measured in supine and standing body positions respectively and PH was defined as a decline from supine to standing was ≥20 mmHg in systolic blood pressures (SBP). The concentrations of blood glucose and plasma insulin were measured to calculate the insulin sensitive index (ISI). Autonomic and peripheral function was determined by measuring the postural heart rates and the conduction speeds of superficial peroneal and communicating branch of peroneal nerves etc respectively. Results　Significant difference (P<0.01) was found in the occurrence of PH in the patients with DM (40%) and those with DMH (67%). The changes of postural blood pressure were more obvious in those with DM+PH and DMH+PH than in those with simple DM (P<0.01). The conduction speeds of newes were significantly lower in those with DMH+PH than with simple DM (P<0.05), but the occurrence of autonomic neuropathy had no difference between the 2 groups. There was no difference in postural heart rate, body mass index and blood glucose levels in fasting and 2 h after meal among the DM, DM+PH and DMH+PH groups. The concentrations of plasma insulin of those with DMH+PH were significantly higher, but their ISI significantly lower than those of the patients with DM respectively (P<0.01). The decline of postural SBP in patients with DMH+PH had a significantly positive correlation with their plasma insulin levels in fasting condition (r=0.689, P<0.01). Conclusion　The patients with DMH are more prone to PH compared with those only with DM and PH damages their peripheral nerves. Most of diabetic patients with PH suffer from obvious IR and hyperinsulinemia, and if with hypertension, the above metabolic disturbances are more severe.

Objective: To evaluate the impact of coronavirus disease 2019 ( COVID-19 ) prevention and control measures in Huzhou on influenza epidemic strength and characteristics in 2020, so as to provide reference for formulating influenza prevention measures. Methods: Using the influenza surveillance data of the national influenza sentinel surveillance system from January 2015 to July 2020, the seasonal characteristics of influenza epidemic were analyzed, the proportion of influenza-like illness cases ( ILI% ) and the positive rate of influenza virus in January to July of 2020 were compared with those of the same period in 2015-2019, in order to evaluate the impact of COVID-19 prevention and control measures. Results : The ILI% and the positive rate of influenza virus in Huzhou were 3.90% and 15.32% during 2015-2019, while were 4.41% and 12.63% from January to July of 2020. The trends of ILI% during 2015-2019 fluctuated similar, but continued to drop since January 2020. The positive rate of influenza virus peaked from December to March in 2015-2019, also peaked from December 2019 to January 2020, but decreased to 0 in March. ILI% was positively correlated with the positive rate of influenza virus ( r=0.682, P<0.05). The growth rates of ILI% from January to July 2020 were 4.75%, -11.27%, 0.68%, 19.84% and 0.92%, compared with the same period of 2015-2019, respectively. The growth rates of ILI% in January 2020 were much higher ( >57.00% ) and from April to July were much lower ( <-33.00% ) . The growth rates of influenza virus positive rate from January to July 2020 were -47.96%, -36.53%, -3.44%, -35.92% and -39.37%, compared to the same period of 2015-2019, respectively. The growth rates of influenza virus positive rate in January 2020 were much higher ( >11.00% ) and from February to March were much lower ( <-61.00% ). Conclusion Since COVID-19 prevention and control measures were implemented in January 2020 in Huzhou, the ILI% and the positive rate of influenza virus in sentinel hospitals decreased significantly.