Objective To construct eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG,and to detect the expression of the plasmid in vitro.Methods hOPG cDNA fragment was extracted from plasmid MGC:29565,amplified by PCR,and inserted into pcDNA3.1(-)vector,then identified by restriction endonuclease digestion and nucleotide sequencing.C2C12 cells were transfected with the plasmid using Lipofectamine 2000.The expression of OPG protein was detected by immunohistochemistry techniques and bone wafer pit assay.Results The constructed recombinant plasmid contained the sequence of hOPG gene.After transfection with the plasmid,active OPG protein could be expressed in C2C12 cells.Conclusion The constructed eukaryotic expression bicistron plasmid vector pcDNA3.1-hOPG can express OPG in vitro,which is the basis for further study on the treatment of bone absorption caused by abnormal activity of osteoclasts.

Objective To observe the CT findings of hypoplasia of cochlear nerve foramen. Methods CT findings of cochlear nerve foramen were observed in 50 volunteers (100 ears)and in 9 cases with hypoplasia of cochlear nerve foramen.The width of the bony canal for the cochlear nerve were measured in both groups.Results The fissure in the cribriform area were seen in every volunteers.The width of the bony canal for the cochlear nerve was (2.32?0.25)mm in normal group.The stenosis of cochlear nerve foramen were shown in 9 cases (10 ears),the lacking of fissure in the cribriform area were seen in 8 ears, while the fissure were shown in 2 cases.The stenosis of internal auditory canal was shown in 4 cases. Abnormality of vestibule and horizontal semicircular canal was shown in one case.The maximum value of the width of the bony canal for the cochlear nerve was 1.5mm,the minimum value was 0.9 mm.Conclusions The typical CT findings of hypoplasia of cochlear nerve foramen were stenosis of cochlear nerve foramen,the lacking of fissure in the cribriform area and cochlear nerve foramen without helix shape.The hypoplasia of cochlear nerve foramen may be a subtype of cochlear dysplasia.

A comparable study were carried out by determination of trace elements on five marine-derived shell traditional Chinese medicine (TCM) (Ostreae Concha, Haliotidis Concha, Margaritifera Concha, Meretricis Concha, and Arcae Concha), which were recorded in the Chinese Pharmacopoeia (2010 version). Seven trace elements in 51 batches of this type of shell TCM were analyzed by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), combined with principal component analysis (PCA) methods. The content of element Se, which exhibited significant differences among different drugs, could be used as a key element to distinguish this type of drugs. Meanwhile, the contents of elements Co, Cu, Mo, and Ba in Haliotidis Concha, Co and As in Margaritifera Concha, Mo and As in Meretricis Concha, Mo, As, and Ba in Arcae Concha, and Zn in Meretricis Concha were relatively stable. In the PCA plot, Arcae Concha and Meretricis Concha could be efficiently distinguished from Ostreae Concha together with Haliotidis Concha, and Margaritifera Concha. The results also showed a correlation with their medicinal function. In conclusion, trace elements in marine-derived shell TCM could not be neglected for their quality control.
Animal Shells ; chemistry ; Animals ; Aquatic Organisms ; chemistry ; Bivalvia ; chemistry ; Mass Spectrometry ; Medicine, Chinese Traditional ; Trace Elements ; analysis

Objective To analyze the alterations of serum protein fingerprint in patients with hypertensive disorder complicating pregnancy(HDCP),screen serum biomarker and establish a diagnostic model of HDCP.Methods Surface-enhanced laser desorption lionization-time of flight-mass spectrometry (SELDI-TOF-MS)technology was used to analyze serum including 25 cases of HDCP patients and 30 cases of age-,gravity-and parity-matched healthy term pregnant women on IMAC3-Cu proteinchip before delivery. Biomarker Wizard and Biomarker Pattern software was used to detect protein peaks significantly different between HDCP and controls,and establish a primary diagnostic model of HDCP.This model was further evaluated by blind test using other 16 parts of serum protein fingerprint.Results Ten protein peaks were significantly different at the molecular range of 2000-50 000(P

OBJECTIVETo summarize the clinical application results of the repair soft tissue defect in contralateral leg with a cross-leg soleus muscle flap pedicle transplantation.

METHODSFrom January 2008 to January 2013, 8 patients with soft-tissue defect in lower leg underwent reconstruction with a cross-leg soleus muscle flap pedicle transplantation (without microvascular anastomoses). There were 7 males and 1 female, aged from 20 to 49 years old with an average of 31.8 years. The operative time after injury was from 2 to 8 weeks with the mean of 46 days. The soleus muscle flap was transposed across to the contralateral leg defect area, then immediate to perform the coverage of the muscle flaps by a meshed split-thickness skin graft. The donor site was closed directly.

RESULTSAll the muscle flaps had survived completely. In one case, recipient area edge had a less exudate from drainage hole everyday, the incision spontaneously was healed after 2 week's changing dressing. Follow-up period ranged form 1.5 to 4 years with an average of 2.5 years postoperatively. The tibia and fibula fractures were healed well. A good contour was achieved at the recipient area. According to LEM standard, 2 cases got excellent results, 5 good and 1 fair.

CONCLUSIONSoleus flap pedicle transplantation is very suitable to repair the soft tissue defect of the injuried leg only one main blood vessel, and can reduce the damage of donor area.

Adult ; Female ; Humans ; Leg Injuries ; surgery ; Male ; Middle Aged ; Muscle, Skeletal ; Soft Tissue Injuries ; surgery ; Surgical Flaps

OBJECTIVETo summarize the clinical application result of the selective nerve root blocks in limited operation of the lumbar spine.

METHODSFrom January 2008 to October 2012,68 patients with lumbar spinal canal stenosis with multiple levels were underwent the selective nerve root blocks in limited operation of the lumbar spine,including 47 males and 21 females with an average age of 56 years old ranging from 45 to 80. After never roots blocks,64 cases were positive for limited operation of the lumbar spine; the other 4 cases were negative and abort the operation.

RESULTSThe nerve roots block operation smoothly and no complications related to the nerve roots block occurred. There was no neurologic injury complication in this study. Follow-up period ranged from 16 to 45 months postoperatively (means, 32 months). The recovery effect was calculated with Macnab scores, the result was excellent in 44 cases, good in 18 cases, poor in 1.

CONCLUSIONOperative treatment for lumbar spinal canal stenosis with multiple levels is focused on the areas causing symptomate neural compression rather than prophylactic decompression at areas of nonsymptomatic disease. Application of selective nerve root blocks can accurately judge the responsible vertebral body and pain source and improve the curative effect of limited operation of the lumbar spine

Aged ; Aged, 80 and over ; Female ; Humans ; Lumbar Vertebrae ; surgery ; Male ; Middle Aged ; Nerve Block ; methods ; Spinal Nerve Roots ; Spinal Stenosis ; surgery

Objective To evaluate the clinical significance of serum level of vascular endothelial growth factor (VEGF),angiopoietin (Ang)-1 and Ang-2 in patients with rheumatoid arthritis (RA).Methods Serum levels of VEGF,Ang-1 and Ang-2 were measured with enzyme linked immunosorbent assay (ELISA).Twenty-one healthy subjects,24 osteoarthritis patients and 82 rheumatoid arthritis patients were included.We defined active and inactive group according to RA disease active score,while early active RA and late active RA group were defined on the basis of disease course.There were 28 early active patients,32 late active patients and 22 inactive patients with rheumatoid arthritis.At the same time 29 RA patients were examined with ultrasound.Synovial hypertrophy (US joint count SH,US index SH),synovial fluid(US joint count SF,US index SF),resistance index and power Doppler signal (US joint PD,TSS) were scored.The correlation was analyzed.We also detected the serum levels of VEGF,Ang-1,Ang-2,ESR,CRP and DAS28 in 25patients with active RA after 3 month regular treatment.We used one-way ANOVA to compare the differences between groups,and Wilcoxon test to compare the differences between before and after treatment.We analyzed the correlation with linear correlation or Spearman rank test.Results The serum level of VEGF [(1285 ±272) pg/ml],Ang-1 [(0.55±0.25)ng/ml] in patients with rheumatoid arthritis were higher than osteoarthritis patients [(934±80) pg/ml,(0.32±0.16) ng/ml] and normal controls [(565±115) pg/ml,(0.24±0.21) ng/ml],and the serum level of Ang-2 [(1.36±0.40) ng/ml] was higher than normal controls [(0.52±0.32) ng/ml].The serum level of VEGF [(1355±194) pg/ml] in early active patients was higher than late active patients [(1096±477) pg/ml] and inactive patients [(862±91) pg/ml].The serum level of Ang-1 in early active patients,late active patients and inactive patients with rheumatoid arthritis had no statistically significant differences.The serum level of Ang-2 in inactive patients [(2.0±2.0) ng/ml] was significantly higher than late active patients [0.9±0.8) ng/ml].The serum level of VEGF was positively correlated with US joint SH,US index SH,US joint PD,and TSS.The serum level of Ang-1 was positively correlated with US joint SH,US joint PD,and TSS.The serum level of VEGF and Ang-1 were negatively correlated with RI.The serum level of Ang-2 was not correlated with US joint SH,US index SH,US joint SF,US index SF,US joint PD,TSS and RI.In the active RA patients,the serum level of VEGF,Ang-1 and Ang-2 was positively correlated with each other.In the inactive RA patients,the serum level of VEGF,Ang-1 and Ang-2 was not correlated with each other.The serum level of VEGF and Ang-1 before treatment was slightly higher than that of after treatment,but the difference was not statistically significant.The serum level of Ang-2 after treatment was significantly higher than that before treatment.ESR,CRP and DAS28 of after treatment were lower than those before treatment.Conclusion The serum VEGF and Ang-1 level could be used as useful index to reflect RA synovial thickening and angiogenesis The serum level of Ang-2 could be used as one of the efficacy indices.They may influence each other,and they may be the key factors that mediate the onset and development of RA angiogenesis and synovial inflammation.

OBJECTIVETo determine the levels of blood spot carnitine and acylcarnitine in children aged 0-15 years by tandem mass spectrometry, offer basic data for evaluating carnitine nutritional status and diagnosing metabolic diseases of organic acid and fatty acid.

METHODSThe concentration of carnitine and acylcarnitines were measured in blood spot by tandem mass spectrometry using underivatized samples. The samples included those from 1376 perinatal neonates, 49 neonates above 1 week of life, 64 children aged up to 1 year and 401 children aged 1 year to 15 years. A few premature infants and low birth weight infants were involved in perinatal neonates without selection. Other samples were taken from mainly outdoor patients for little surgical preoperative examination. Patients suffering from fever, diarrhea, liver disease, severe fat-metabolic diseases were excluded from this study.

RESULTSThe concentrations of carnitine (C(0)); short-chain acylcarnitines (SC-AC), including acetyl (C(2)), propionyl (C(3)), malonyl (C(3)DC), butyryl (C(4)), methylmalonyl (C(4)DC), isovaleryl (C(5)), glutaryl (C(5)DC); middle-chain acylcarnitines (MC-AC), including hexanoyl (C(6)), hexanediol (C(6)DC), octylenoyl (C(8:1)), octanoyl (C(8)), decadienoyl (C(10:2)), decanoyl (C(10:1)), decanoyl (C(10)); total carnitine and acylcarnitines (TCAC)were lower in neonate, highest in 1-3 months of age, higher in 6-12 months of age, and kept at the same level between 2 and 15 years of age. The concentrations of total long-chain acylcarnitines (LC-AC), including lauren (C(12:1)), lauroyl (C(12)), tetradecanoyl (C(14:1)), tetradecanoyl (C(14)), 3-hydroxy-tetradecanoyl (C(14)OH), hexadecenoyl (C(16:1)), hexadecanoyl (C(16)), 3-hydroxy-hexadecanoyl (C(16)OH), 3-hydroxy-hexadecanoyl (C(16:1)OH), octadecadienoyl (C(18:2)), octadecenoyl (C(18:1)), octadecanoyl (C(18)), 3-hydroxy-octadecenoyl (C(18:1)OH), and 3-hydroxy-octadecanoyl (C(18)OH) were the highest in neonate, decreased gradually, and kept the same level between 2 and 15 years of age. The concentrations of C(0) (23.387 ± 7.702) µmol/L, (30.064 ± 8.252) µmol/L, (25.021 ± 6.630) µmol/L, of LC-AC (4.998 ± 1.557) µmol/L, (2.854 ± 0.821) µmol/L, (2.459 ± 0.553) µmol/L, of TCAC (43.497 ± 12.632) µmol/L, (49.013 ± 12.497) µmol/L, (39.656 ± 9.257) µmol/L were significantly different among the groups of neonate, up to 1 year and above 1 year (P < 0.01). The concentrations of C(0) (24.115 ± 7.715) µmol/L and TCAC (43.65 ± 5.252) µmol/L in perinatal male neonates were higher than that (22.696 ± 7.246) µmol/L, TCAC (41.90 ± 5.038) µmol/L in female neonates. The C(0)/TCAC ratio of neonatal group (54.0% ± 7.1%) was significantly lower than that in the children group (62.1% ± 6.1%, P < 0.05), LC-AC/TCAC (33.5% ± 6.0%), MC-AC/TCAC (1.3% ± 0.3%), SC-AC/TCAC (11.6% ± 2.5%)ratios of neonatal group were higher than that of children group respectively (30.1% ± 4.9%; 0.9% ± 0.6%; 6.5% ± 2.3%, P < 0.05).

CONCLUSIONSConcentrations and profiles of carnitine and acylcarnitines change significantly during the first year of life, the age should be considered as a factor when evaluating carnitine nutritional status and diagnosing metabolic diseases of organic acid and fatty acid. Concentrations of carnitine and acylcarnitines were a little higher in male neonates than in female.

Adolescent ; Carnitine ; analogs & derivatives ; blood ; Child ; Child, Preschool ; Female ; Humans ; Infant ; Infant, Newborn ; Male ; Tandem Mass Spectrometry ; methods

Six 89bp primers were designed on the base of the cDNA sequence encoding the human leptin reported on the NCBI. The synthetic gene with 464bp encoding rhLep was obtained by SOE ( splicing by overlap extension) PCR. The expression vector pET22b(+)/rhLep was constructed and transformed into E. coli BL21 (DE3). The rhLep protein was expressed as inclusion bodies with the yield of more than 50% of total bacterial proteins after IPTG induction. The rhLep protein, which has a molecular weight about 16kD, was purified by Ni2+ affinity chromatography column and identified by SDS-PAGE. The MTT Assay shows that rhLep promotes EC304 cells growth at the low concentration of 10ng/mL to 30 ng/mL, and rhLep appears cytotoxic to EC304 cells with the high dose of 50ng/mL to 225ng/mL. The viability of EC304 cells decreases to 1.2% with the concentration of 225ng/mL of rhLep. The massive apoptosis of rhLep on EC304 cells is observed by AO-staining under fluorescent microscope. All these results would lay the foundation for the further study of its biological functions in vitro and in vivo.
Apoptosis ; drug effects ; Escherichia coli ; genetics ; Humans ; Leptin ; genetics ; pharmacology ; Recombinant Proteins ; biosynthesis ; isolation & purification ; pharmacology

Objective To explore the relationship between social participation and needs for rehabilitation of the disabled in Guangdong Province and to make a proposal for developing the rehabilitation strategies. Meth-ods The data of the Second National Sample Survey of Disabled Persons in Guangdong Province was used in this study. Ranked data analysis was made with the sub-items of the social participation assessment and the main needs of the disabled individuals. Results Significantly statistical differences were revealed with regard to the constitu-ent ratio of needs for rehabilitation services among people with different degrees of difficulties in social participation caused by hearing and visual impairments as well as physical and mental disabilities. No significant difference was found in terms of the constituent ratio of rehabilitation needs among those with difficulties in speech and those with psychiatric diseases. The major rehabilitation needs focused on medical service, assistive apparatus support and functional trainings. Conclusions The rehabilitation needs were different among different categories of disabled persons. Rehabilitation services should be provided accordingly.