Objective To investigate the proliferation and collagen secretion of transplanted human fibroblasts.Methods The solution containing human fibroblasts(2?1010L-1)was prepared and 1 mL was injected into the dermis of BALB/CNU nude mice.Animals were killed by the end of the 1st,2nd and 3rd month after injection.The dermis in the injected area was taken out and stained with HE.Immunohistochemical staining for type I and type Ⅲ collagen was performed at the same time.Results Mitosis was observed by the end of the 1st,2nd and 3rd month.The concentration of type I and type Ⅲ collagen in the extra cellular matrix increased with the passing of time.Conclusion Transplanted human fibroblasts can proliferate automatically in the dermis of nude mice and manufacture the type I and type Ⅲ collagen in situ.Long period of survival and secretion will make it possible for fibroblasts to become promising option to correct minimal tissue defects.

Objective To compare 99Tcm-MIBI MPI with delayed enhancement MRI (DE-MRI) in patients with idiopathic dilated cardiomyopathy (IDCM). Methods Forty patients with IDCM were included. They underwent both rest 99Tcm-MIBI myocardial perfusion imaging and DE-MRI within 7 days. 99Tcm-MIBI MPI was performed to identify diffuse or segmental abnormal perfusion patterns including reduced or defect perfusion segments. DE-MRI images were divided into 4 categories: no delayed enhancement, septal, subendocardial and transmural delayed enhancement, x2 test was used for data analysis. Results Diffuse and segmental perfusion abnormality on 99Tcm-MIBI MPI were found in 19 (47.5%) and 21 (52.5%)patients respectively, while DE-MRI enhancement was simultaneously found in 5 patients of the former (5/19, 26.3%) and 18 (18/21, 85.7%) of the latter (x2 =14.401, P＜0. 001). For those (n=18) with both segmental perfusion abnormality and DE-MRI enhancement, the number of segments of the 4 DE-MRI respectively. A significant difference was found in the DE-MRI enhancement categories between normal and defect perfusion segments (x2 = 29. 183, P ＜0.001 ) and between reduced and defect perfusion segments as well (x2 =25. 110, P＜0. 001). Conclusions Both diffuse and segmental perfusion abnormalities on 99Tcm-MIBI MPI can be found in patients with IDCM. DE-MRI enhancement is more frequently found in patients with segmental perfusion abnormality.

BackgroundRecent researches suggested that properties of neurons in the lateral geniculate neuron (LGN) may represent an important neural limitation on the development of basic spatial and temporal vision,and even binocular rivalry.However,previous studies on the properties of spatiotemporal frequency tuning of LGN were rather concentrated on a monkey or cat,whereas little is known about rat.ObjectiveThis study was to examine the development of spatiotemporal frequency tuning in rats LGN.MethodsTwenty Wistar rats were collected and divided into 14-16 day,20-22 day,27-30 day and 60 day groups according to the different ages after their eyes opened and 5 rats were assigned for each group.Extracellular single neuron recording was carried out in the rats to study the spatio-temporal receptive field properties of neurons in LGN by sinusoidal gratings visual stimuli.Dynamic changes of the spatio-temporal receptive field properties of neurons in LGN with the development of Wistar rats were evaluated.ResultsThe differences between band-pass and low-pass distribution of temporal frequency or spatial frequency of rat LGN were not statistically significant (x2 =0.68,0.47,P＞0.05 ).The optimal temporal frequency of receptive field in rat LGN went up to the maximum value until 60 day in Wistar rats.The mean optimal temporal frequencies of neurons in the four different age groups were ( 2.5 ± 1.3 ),( 2.6± 1.2),(2.6± 1.1 ) and ( 3.6± 1.1 ) Hz with significant differences among the 4 groups (F=4.53,P＜0.05 ),and those in the 14-16 day group,20-22 day group,27- 30 day group were significantly lower than in the 60 days group ( q =4.43,4.10,4.03,P ＜ 0.05 ).No significant differences were seen among the 14-16 day group,20-22 day group and 27-30 day group ( P＞0.05 ).The optimal spatial frequency values in the four groups were ( 0.04 ± 0.04 ),( 0.04 ± 0.03 ),( 0.05 ± 0.03 ),( 0.05 ± 0.04 ) cpd,respectively without statistical difference among them ( F =0.58,P ＞ 0.05 ).The temporal and spatial bandwidth values in the various age groups were not statistically significant among the four groups ( F =0.37,1.22,P＞0.05). Conclusions The development of temporal and spatial frequency characteristics of the rat LGN receptive field may be related to its functional visual pathway.

To provide accurate information on geographic distribution of crude drug Sailonggu in the plateau, we identified zokor species (Eospalax spp.) in Qinghai-Tibet Plateau using molecular methods. Based on the mitochondrial cytochrome B (cytb) gene sequences, we then extracted haplotypes from these sequences and reconstructed phylogenetic trees for the haplotypes using both maximum likelihood (ML) and Bayesian inference (BI) methods. Based on the trees, the species of each sample were determined. Five hundred and three samples from 35 populations were sequenced and their whole cytb sequences (1140 bp) were obtained. From these sequences 150 haplotypes were detected, in which, 126 were Eospalax baileyi, 20 were E. cansus, and 4 were E. smithi of the 35 populations, 28 were E. baileyi type, 5 were E. cansus type, and the remaining 2 were mixed of E. baileyi + E. cansus (DT2) and E. baileyi + E. smithi (ZN3). The results showed that, the regions around the Qinghai lake and near the upper stream of Yellow River started at Guide could be viewed as the producing area of authentic Sailonggu, and also, the cytb gene is a powerful molecular marker to determine the species of zokors as well as for the authentication of geographic distribution of Sailonggu.
Animals ; Bone and Bones ; metabolism ; Haplotypes ; Medicine, Tibetan Traditional ; Phylogeny ; Rodentia ; classification ; genetics

OBJECTIVETo observe the differential effect of joint ultrasound on the syndrome differentiation of rheumatoid arthritis (RA) by observing the high frequency ultrasound performances among inactive stage and different syndromes in active stage.

METHODSTotally 83 RA patients in the active stage were assigned to the dampness heat syndrome group (DHS, 59 cases)and the cold dampness syndrome group (CDS, 24 cases) according to Chinese medicine (CM) syndrome typing. Besides, 20 RA patients in the remission stage were recruited as the control group (abbreviated as the remission group). By using high frequency ultrasound and power Doppler ultrasound technology, a comparative observation of synovitis, tenosynovitis, synovial blood flow, and bone erosion in the 2nd-5th metacarpophalangeal (MCP) joints, proximal interphalangeal (PIP) joints, wrist joints, knee joints, the second and the fifth metatarsophalangeal (MTP) joints (a total of 24 joints) was performed in all patients. Correlation analyses were performed between the ultrasound performance, laboratory indices, and the disease activity. Ultrasound data of each RA patient were analyzed by their total scores. Χ2 test was used for enumeration data. The measurement data was expressed as x ± s. One-way ANOVA was used for data of normal distribution, while non- parametric test was used for data of non-normal distribution. Correlation analysis of two variables was performed for clinical indicators and ultrasound indicators. Its significance was detected using Pearson correlation.

RESULTSCompared with the remission group, the severity degree of synovitis, tenosynovitis, synovial blood flow, and bone erosion significantly increased in the DHS group (P < 0.01). There was statistical difference in ESR, CRP, anti-CCP, DAS28 score, and the positive rate of RF (P < 0.05, P < 0.01). There was statistical difference in the severity degree of synovitis and synovial blood flow, and DAS28 score in the CDS group (P < 0.05). Compared with the CDS group, there was statistical difference in the four ultrasound indices (P < 0.05, P < 0.01), ESR, CRP, anti-CCP, DAS28 score, and the positive rate of RF in the DHS group (P < 0.05, P < 0.01). There was no statistical difference in G, IgG, IgA, or IgM among the three groups (P > 0.05). There existed positive correlation between ESR and the synovitis degree, synovial blood flow, and bone erosion in the DHS group (r = 0.444, 0.397, 0.486, P < 0.05).There existed positive correlation between ESR and the synovitis degree, bone erosion, and synovial blood flow in the DHS group (r = 0.378, 0.270, P < 0.05). There existed positive correlation between the DAS28 score and the synovitis degree and synovial blood flow in the DHS group (r = 0.304, 0.351, P < 0.05).

CONCLUSIONSThe inflammation degree was the most severe in RA patients of DHS. High frequency ultrasound could provide better evidence for Chinese medical syndrome differentiation of RA patients.

Arthritis, Rheumatoid ; diagnostic imaging ; Humans ; Medicine, Chinese Traditional ; Metacarpophalangeal Joint ; ultrastructure ; Syndrome ; Synovitis ; diagnostic imaging ; Ultrasonography

Objective To investigate the value of the Wells score and D-dimer assay in assisting pulmonary perfusion imaging (PPI) for the diagnosis of acute pulmonary embolism (APE). Methods One hundred twenty-one patients with suspected APE were studied from January, 2006 to December, 2008. All patients underwent the Wells score, the quantitative D-dimer assay, chest X-ray photography, and PPI. The diagnostic sensitivity, specificity, positive predictive value and negative predictive value of PPI with the assistance of Wells score and D-dimer assay were calculated. Results Fifty (41.3%) patients were diagnosed with APE. PPI combined with chest X-ray photography (Q/X scan) showed positive results in 49 patients. The sensitivity, specificity, positive predictive value and negative predictive value of the Q/X scan were 86.0% (43/50), 91.5% (65/71), 87.8% (43/49) and 90.3% (65/72), respectively. With assistance of Wells score >4 and D-dimer≥0. 5 mg/L, Q/X scan had a positive predictive value of 100.0% (29/29), for patients with Wells score ≤4 and D-dimer<0.5 mg/L, the negative predictive value for Q/X scan was 100.0% (41/41). Conclusion Combined with Wells score and D-dimer assay, PPI can make accurate diagnosis of APE.

Background: There have been few reports of mutations in the beta-myosin heavy chain(MYH7)gene in hypertrophic cardiomyopathy(HCM),which is associated with sudden cardiac death caused by HCM.This study aimed to screen the mutation sites in the sarcomeric gene MYH7 in Chinese patients with HCM.We also p1anned to analyze the pathogenicity of the mutation site as well as its significance in clinical and forensic medicine.Methods: From January 2006 to June 2017,autopsy cases were collected from the Department of Pathology,the Affiliated Hospital of Qingdao University.The experiment was to detect MYH7 gene status in formalin-fixed paraffin-embedded tissues from 18 independent autopsy cases who suffered HCM related sudden death(fatal HCM)and 20 cases without cardiomyopathy.Common mutation exon fragments of MYH7 gene were amplified by polymerase chain reaction.The end-of-deoxygenation method and gene cloning method were further performed to analyze the mutation sites.Homologous comparison among mutant sites was conducted using BLAST online database.Results:The 1336th nucleotide of MYH7 gene at exon 14 was converted from T to G in one HCM case,resulting in the conversion of threonine(Thr)at position 446 to proline(Pro).In another case,the 1402th nucleotide at exon 14 was converted from T to C,resulting in the conversion of phenylalanine(Phe)at position 468 to leucine(Leu).Homologous comparison results showed that the two amino acid residues of Thr446 and Phe468 are highly conserved among different species.Conclusions: Our results showed fatal HCM harbored mutations of Thr446Pro and Phe468Leu in the MYH7 gene.It is significant for clinical and forensic medicine to further explore the functions and detailed mechanisms of these mutations.

OBJECTIVETo investigate the treatment method for collapse separation fractures of lateral tibial condyle.

METHODSFrom January 2003 to October 2008, 45 patients with collapse separation fractures of lateral tibial condyle were treated by surgical method, including 27 males and 18 females,ranging in age from 15 to 76 years,with an average of 37 years. The duration from injury to operation ranged from 1 to 14 days. All of the patients preoperatively underwent X-ray examinations in order to accurately understand the extent of fracture and fracture collapse. All of the 45 fractures with collapse more than 2 mm and widen more than 5 mm were selected for surgical treatment. The lateral surgical approach was used to expose the lateral margin of the lateral meniscus first, and then the reduction of the lateral meniscus and lateral condyle were achieved. The results after operation were evaluated by the Rasmussen criteria, as well as the complications of the knee joint pain, the joint activity, the infection, deep venous thrombosis and recurrent fracture collapse.

RESULTSPostoperative X-ray showed anatomical reduction was achieved in 31 patients, 14 patients got nearly anatomical reduction. The plate and screw position were satisfactory. Forty-two patients were followed up,and the duration ranged from 6 to 12 months, with an average of 10 months. Fracture re-displacement,postoperative infection and deep venous thrombosis did not occur after the lifting of external fixation. All the patients had pain disappeared about 10 days after operation,and had no vertical axis percussion tenderness at 3 weeks after operation. The X-ray showed fractures was healed in all patients at 3 months after operation. Joint activity Rasmussen assessment criteria results: 37 patients got an excellent result, 7 good, 1 poor.

CONCLUSIONThe incarceration of lateral meniscus and the collapsed bone fragments are two factors which make the reduction of the lateral tibial condyle difficult.

Adolescent ; Adult ; Aged ; Female ; Fracture Fixation ; methods ; Humans ; Male ; Middle Aged ; Tibial Fractures ; physiopathology ; surgery

Objective To investigate the effects of intrathecal dexmedetomidine administration on the development of morphine tolerance and spinal inflammatory responses.Methods Thirty-three male SD rats weighing 180～200 g were randomly divided into 3 groups (n=11):Saline group (group NS),Morphine group (group M) and Dexmedetomidine group (group Dex).Animals of group NS were intrathecally injected with 10 tμL of saline daily for seven days;Animals in group M were intrathecally injected with 15 μg of morphine daily for seven days;Animals in group Dex were intrathecally injected with a mixture of 15μg morphine and 1.5 μg dexmedetomidine daily for seven days.At 1,3,5 and 7 day of intrathecal injection,hot water tail-flick test were used to evaluate analgesic response to thermal stimuli.After the last episode of behavioral test,Western blot analysis was applied to determine the protein levels of Iba-1 (microglial marker),IL-1β,TNF-a and phospho-p38MAPK (p-p38) in the spinal cord.In addition,microglia in the spinal cord was immuno-stained with anti-Iba-1 antibody and the densities of microglia were calculated.Results In group M and Dex,the values of maximal possible effect (MPE) in tail-flick test decreased gradually along with repeated morphine administration (P＜0.05).Compared with group NS,the values of MPE in tail-flick test at 1,3,5 and 7 day of morphine tolerance were higher in group M (P＜0.05).Compared with group M,the values of MPE in tail-flick test at 3,5 and 7 day of morphine tolerance were higher in group Dex (P＜0.05).Compared with group NS,the spinal protein levels of Iba-1,IL-1L TNF-α and p-p38 as well as the density of Iba-1 positive cells in group M were increased (P＜0.05).However,Compared with group M,the of Iba-1,IL-1β,TNF-α and p-p38 as well as the density of Iba-1 positive cells were decreased(P＜0.05).Conclusion Intrathecal dexmedetomidine administration can attenuate morphine tolerance by inhibiting microglia-mediated inflammatory responses in the spinal cord.

Objective To establish an automatic synthesis method for 11C-carfentanil (CFN) as an novel opiate receptor radioligand and study its biodistribution in rats. Methods 11C-Triflate-CH3 was bubbled into 0.5 mg precursor desmethyl-CFN (which was dissolved in 0.15 ml DMSO) to generate 11C-CFN in a V-tube at room temperature. Sep-Pak C2 column was used for purification of 11C-CFN, which was eluted by 3ml binary system aqueous solution, 10 ml water thrice, and then I ml ethanol. The biodistribution (% ID/g) of 11C-CFN in SD rats was studied. SPSS 13.0 was used for statistical analysis. Non-normal distribution data were analyzed using nonparametric test. Results The synthesis time for 11C-CFN was 20 min (end of bombardment, EOB). The synthesis yield was (35.5 ± 2.2) % on average (n = 12, uncorrected)with the radiochemical purity over 98%. Biodistribution study in rats showed that the tracer had a high brain uptake, rapid blood clearance, and a metabolic pathway via liver and kidney. The highest tracer uptake was in thalamus (4.26 ± 0.89) % ID/g and striatum (4.05 ± 1.08) % ID/g at 5 min after injection, followed by cerebral cortex (2.63±0.89) %ID/g, pons (2.26 ±0.57) % ID/g, hippocampus (2. 17 ±0.55) %ID/g and cerebellum (2. 15 ±0.39) %ID/g. Conclusions The automatic synthesis of 11C-CFN is fast and reliable, and this radioligand can be used for opiate receptor imaging.