To study the MEN1 gene mutations in a multiple endocrine neoplasia type 1 ( MEN 1 ) family,and determine the possible mechanism of disease induced by the mutations.Genomic DNA was isolated from peripheral blood leukocytes and the MEN1-related tumor tissues of the patient and the family members,then the coding exons and exon/intron boundaries of the MEN1 gene were amplified by polymerase chain reaction (PCR) and sequenced.Subclone sequencing was performed to identify the heterozygosity.Further immunohistochemistry was performed to observe menin protein expression in the tumor tissues.We identified a heterozygous deletion mutation of intron 9 ( IVS9+ 1_11 delGTGAGGGACAG) in the proband and two family menbers.We also demonstrated for the first time that the expression of menin protein is absent in the parathyroid adenoma tissue.The heterozygous mutation in the initial of intron 9,IVS9+ 1_11 delGTGAGGGACAG is a new type of MEN1 gene mutations in China.This mutation may produce an aberrant splicing of MEN1 mRNA,generating easily degradation and loss of expression of menin protein and resulting eventually in the disease.

The fine structure of cells in the trigeminal mesencephalic (Me5) nucleus in the rat was studied by transmission electron microscopy. This nucleus located in the mid-brain and the rostral portion of the pons, most neurons in the caudal part of Me5 nucleus were sufficiently large (40~50micrometer) and appeared as round-shaped unipolar cells. The Me5 neurons had a large, round, centrally located nucleus, and their cytoplasm contained numerous mitochondria, dense granular endoplasmic reticulum, abundant Golgi apparatus, groups of free ribosomes, some heterogeneous particles and neurofilaments. Cell surfaces were studded irregularly with small spinouts processes, and they contained a few fine irregularly arranged neurofilaments and some granular endoplasmic reticulum. Boutons contacting the soma of Me5 neurons were defined as axosomatic synapses and bostons contacting dendrites located between the Me5 neurons were defined as axodendritic synapses. Based on differences in bouton and vesicle morphology, the four synaptic bouton types were identified. 1. Asymmetrical as well as symmetrical synapses with small round vesicles. 2. Asymmetrical synapses with pleomorphic admixture contained predominantlyspherical vesicles. 3. Symmetrical synapses with pleomorphic vesicles of flattened, spherical and dense-core vesicles. 4. Asymmetrical as well as symmetrical synapses with heterogeneous and large dense-core vesicles. Synaptic boutons containing round vesicles and large dense-core vesicles were most frequently observed.
Animals ; Carisoprodol ; Cytoplasm ; Dendrites ; Endoplasmic Reticulum, Rough ; Golgi Apparatus ; Microscopy, Electron, Transmission ; Mitochondria ; Neurons ; Pons ; Presynaptic Terminals ; Rats* ; Ribosomes ; Synapses

The maturation pattern of the calcium binding proteins parvalbumin (PV) and calbindin-D28K (CB) from the day of birth, postnatal day 0 (P0) to 30 days (P5, P10, P15, P20, P30) and adult was studied in the rat amygdala using immunohistochemistry. PV and CB immunoreactivities in the amygdala of the rat showed very different patterns during postnatal development. The first PV-immunoreactive neurons appeared in the cortical amygdaloid nuclei and the basolateral amygdaloid nucleus at P5, and then in the lateral amygdaloid nucleus and the basomedial amygdaloid nucleus at P10. Adult patterns of PV-immunoreactive neurons were reached at P20. In contrast, CB-immunoreactive neurons were already found at birth in all amygdaloid nuclei except the intercalated nucleus. The intensity and number of immunoreactivity of CB-containing neurons increased during the first 10 days of postnatal life but dramatically decreased at P15. Mature patterns CB-immunoreactive neurons were achieved at P20. These two calcium binding proteins exhibited a non-homogeneous distribution in the adult amygdala, PV-immunoreactive neurons were mainly localized in the basolateral nuclear group but not in the medial amygdaloid nucleus, the cental amygdaloid nucleus and the intercalated nucleus. In contrast, CB-immunoreactive neurons were distributed in almost all amygdaloid nuclei except the intercalated nucleus. The present results showing different postnatal maturation patterns such as time of appearance, the number and distribution of immunoreactive cells suggest that PV and CB may play a different functional role during the postnatal development of the amygdala.
Adult ; Amygdala ; Animals ; Calbindin 1 ; Calbindins* ; Calcium-Binding Proteins ; Humans ; Immunohistochemistry ; Neurons* ; Parturition ; Rats*

PURPOSE: The purpose of this study was to evaluate the 15.5 year long term survival rate of the Precoat femoral stem. MATERIALS AND METHODS: We reviewed the results of 105 primary hybrid total hip replacements (98 patients) that were performed by one surgeon between October 1990 and August 1995 using a cemented polymethyl-methacrylate coated femoral prosthesis (Precoat) and contemporary cementing techniques. Thirty four patients (34 hips) died and seventeen patients (17 hips) were lost to follow-up. Forty seven patients (54 hips) were available for clinical follow-up, with an average follow-up period of 15.5 years (range: 8.4 to 18.3 years). The average age of the patients at the time of the index operation was 46 years (range: 22 to 67 years). There were 32 male patients (37 hips) and 15 female patients (17 hips). RESULTS: For the acetabular component, 15 hips (27.8%) were revised for cup loosening and isolated liner exchange was performed in 12 hips (22.2%) for liner wear and osteolysis. For the femoral component, 12 hips (22.2%) were revised due to aseptic loosening. Of these twelve hips, 3 hips had Grade B cement mantles and 9 had Grade C cement mantles. The clinical results of the 54 retained hips were good or excellent in 52 hips (96.3%) with the average Harris hip score being 88 points (range: 72 to 96 points). CONCLUSION: The mean 15.5 years' survival rate of the Precoat cemented femoral stem was 78%. We think that there were several factors for the failure of femoral stem fixation, including age, physical activity and the body weight, as well as the stem design and the surgical technique had an influence on the stem's survival.
Arthroplasty ; Arthroplasty, Replacement, Hip ; Body Weight ; Chimera ; Female ; Follow-Up Studies ; Hip ; Humans ; Lost to Follow-Up ; Male ; Motor Activity ; Osteolysis ; Prostheses and Implants ; Survival Rate

Objective To build a high sensitive method to detect HIV-1 P24 antigen quantitatively by Immuno-rotary Biosensor (IRB) based on F0F1-ATPase. Methods The immuno-rotary biosensor based on F0F1-ATPase in chromatophores for detecting HIV-1 P24 was assembled by label F0F1-ATPase chromatophores with the biotin-labled HIV-1 P24 antibody through the streptavidin-biotin-β antibody and the pH sensitivity of fluorescence F1300. Then it's sensitivity and specificity was tested by the prokaryotic expressed HIV-1 P24 antigen. Results Immuno-rotary biosensor based on F0F1-ATPase in chromatophores can detect P24 antigen sensitivily, the lowest level that could be detected was 0.1 fg/ml. Conclusion IRBbased system was successfully assembled and used for the detection of P24 antigen, the rapid and sensitive technique will be useful for detecting HIV-1 P24 antigen quantitatively.

This study investigated the anti-cancer potential of a near-infrared fluorescence (NIRF) molecule conjugated with Cetuximab (Cetuximab-NIRF) in six-week-old female BALB/c athymic (nu+/nu+) nude mice. A431 cells were cultured and injected into the animals to induce solid tumors. Paclitaxel (30 mg/kg body weight (BW)), Cetuximab (1 mg/kg BW), and Cetuximab-NIRF (0.25, 0.5 and 1.0 mg/kg BW) were intraperitoneally injected twice a week into the A431 cell xenografts of the nude mice. Changes in BW, tumor volume and weight, fat and lean mass, and diameter of the peri-tumoral blood vessel were determined after two weeks. Tumor volumes and weights were significantly decreased in the Cetuximab-NIRF (1 mg/kg BW) group compared with the control group (P < 0.001). Lean mass and total body water content were also conspicuously reduced in the Cetuximab-NIRF (1 mg/kg BW) group compared with the vehicle control group. Peri-tumoral blood vessel diameters were very thin in the Cetuximab-NIRF groups compared with those of the paclitaxel group. These results indicate that the conjugation of Cetuximab with NIRF does not affect the anti-cancer potential of Cetuximab and NIRF can be used for molecular imaging in cancer treatments.
Animals ; Blood Vessels ; Body Water ; Body Weight ; Cetuximab* ; Female ; Fluorescence* ; Heterografts ; Humans ; Mice ; Mice, Nude ; Molecular Imaging ; Paclitaxel ; Tumor Burden ; Weights and Measures

OBJECTIVETo evaluate the status of Haemophilus influenza type b(Hib) on death cases of children from community-acquired pneumonia (CAP) and to estimate the value of direct in-situ polymerase chain reaction (ISPCR) on diagnosis of children CAP, pathogenically.

METHODSOrdinary PCR, Southern blot and direct ISPCR were applied and compared in detecting Hib in 100 paraffin-embedded lung tissues of autopsy children died of CAP.

RESULTSNo major difference on the detection rate of Hib between 50-60s and 80s-2002 was found. The detection rate of Hib by direct ISPCR was higher than the other two methods. By Southern blot, Hib was identified from 8 out of 100 samples (8%), including 4 out of 56 in 1950-60s (7.1%) and 4 out of 44 (9.1%) (chi2 = 0.084, P>0.05) in 1980s-2002. By ISPCR, Hib was identified from 17 out of 100 samples (17%), including 8 out of 56 in 1950-60s (14.3%) and 9 out of 44 (20.5%) with chi2 = 0.665, P > 0.05, in 1980s-2002. Positive cases diagnosed by both Southern blot and ISPCR were 7%.

CONCLUSIONHib was one of the main bacterial pathogens causing CAP and deaths among children. Direct ISPCR was prefertable to be used in pathogenic diagnosis on children pneumonia, in terms of its sensitivity, specificity and localization.

Age Factors ; Autopsy ; Blotting, Southern ; Child, Preschool ; Community-Acquired Infections ; microbiology ; pathology ; Female ; Haemophilus influenzae type b ; genetics ; physiology ; Humans ; Infant ; Lung ; microbiology ; pathology ; Male ; Pneumonia ; microbiology ; pathology ; Polymerase Chain Reaction

This study was investigated the change of concentration and toxicity of thermally cross-linked superparamagnetic iron oxide nanoparticles (TCL-SPION) on tissues of Sprague-Dawley rats. TCL-SPION at the dose of 15 mg/kg body weight was intravenously injected into the tail vein of the male Sprague-Dawley rats. The fate of TCL-SPION in serum, urine and tissues was observed during 28 days. Serum iron level was maximal at 0.25 h post-injection and gradually declined thereafter. In addition, the sinusoids of liver and the red pulp area of spleen were mainly accumulated iron from 0.5 h to 28-day post-injection. In kidney, iron deposition was detected in the tubular area until 0.5 h after injection. Malondialdehyde concentration in the liver slightly increased with time and was not different with that at zero time. In the liver and spleen, TNF-alpha and IL-6 levels of TS treated with TCL-SPION were not different with those of the control during the experimental period. From the results, TCL-SPION could stay fairly long-time in certain tissues after intravenous injection without toxicity. The results indicated that TCL-SPION might be useful and safe as a contrast for the diagnosis of cancer or a carrier of therapeutic reagents to treat diseases.
Animals ; Body Weight ; Diagnosis ; Humans ; Indicators and Reagents ; Injections, Intravenous ; Interleukin-6 ; Iron* ; Kidney ; Liver ; Male ; Malondialdehyde ; Nanoparticles* ; Rats ; Rats, Sprague-Dawley* ; Spleen ; Tumor Necrosis Factor-alpha ; Veins

PURPOSE: The aim of this study is to analyze the clinical results of using the technique of rotator cuff repair without parting the biceps long head from the glenoid for large or massive tear of the rotator cuff. MATERIAL AND METHODS: Form January 2005 to January 2007, we performed the arthroscopic biceps repair with incorporating suture to the rotator cuff for 21 patients with large or massive rotator cuff tear. The mean follow up period was 23 months (range: 6-48months). The number of males and females was 9 and 13, respectively. The age distribution ranged from 47 to 73 years with a mean age of 60.3 years. We compared the preoperative score with the postoperative scores using the University of California Los Angeles (UCLA) score, the shoulder index of the American Shoulder and Elbow Surgeons (ASES) and a simple shoulder test (SST). RESULTS: The improvement in the VAS, ASES and the UCLA and SST scores was statistically significant at the final follow up (average follow-up 23 months) (p>0.05). Two of nine cases were found to have partial tear with continuity but seven cases were found to have complete tear according to the ultrasonography and MRI. CONCLUSION: The technique of rotator cuff repair without parting the biceps long head from the glenoid for large or massive tear of the rotator cuff is considered to be recommendable.
Age Distribution ; California ; Elbow ; Female ; Follow-Up Studies ; Head ; Humans ; Los Angeles ; Male ; Rotator Cuff ; Shoulder ; Sutures ; Tendons

Free Cy5.5 dye and Cy5.5-labeled thermally cross-linked superparamagnetic iron oxide nanoparticles (TCL-SPION) have been routinely used for in vivo optical imaging. However, there is little information about the distribution and accumulation of free Cy5.5 dye and Cy5.5-labeled TCL-SPION in the tissues of mice. Free Cy5.5 dye (0.1 mg/kg body weight) and Cy5.5-labeled TCL-SPION (15 mg/kg body weight) were intravenously injected into the tail vein of ICR mice. The biodistribution and accumulation of the TCL-SPION and Cy5.5 were observed by ex vivo optical imaging and fluorescence signal generation at various time points over 28 days. Cy5.5 dye fluorescence in various organs was rapidly eliminated from 0.5 to 24 h post-injection. Fluorescence intensity of Cy5.5 dye in the liver, lung, kidney, and stomach was fairly strong at the early time points within 1 day post-injection. Cy5.5-labeled TCL-SPION had the highest fluorescence density in the lung at 0.5 h post-injection and decreased rapidly over time. Fluorescence density in liver and spleen was maintained over 28 days. These results suggest that TCL-SPION can be useful as a carrier of therapeutic reagents to treat diseases by persisting for long periods of time in the body.
Animals ; Carbocyanines/*pharmacology ; Ferric Compounds/*pharmacology ; Fluorescent Dyes/*pharmacology ; Kinetics ; Male ; Mice ; Mice, Inbred ICR ; Nanoparticles/*metabolism ; Time Factors ; Tissue Distribution