Matrix effect on the determination of Potency in Recombinant Coagulation Factor Ⅷ for injection / 中国输血杂志

【Objective】 To investigate the matrix effect on the determination of potency in Recombinant Human Coagulation Factor Ⅷ for Injection (rFⅧ). 【Methods】 Two different detection matrices were used to establish two methods for detecting the potency in Recombinant Human Coagulation Factor Ⅷ for Injection. And the matrix effect on the determination of potency was determined, including specificity, linearity, repeatability, accuracy and intermediate precision. 【Results】 As to the specificity, the recoveries of the two substrates at high vs low concentration level were 112% and 110% vs 104% and 109%, respectively. As to the linearity, in the range of (0.125-1.000) IU/mL, the correlation coefficient between concentration and coagulation time of standard/ sample was higher than 0.99. As to the accuracy/repeatability, the recoveries of two matrices was 104% and 102%, and RSD was 2.4% and 1.9%. As to the intermediate precision, personnel factor of two matrices was 0.72 and 0.23, date factor was 0.79 and 0.85, and RSD(for 12 times) was 4.2% and 3.0%. Comparison of two matrices was as follows: Deviation in test results of 6 batches of rFⅧ was all lower than 5%. There was no significant difference between two matrices. 【Conclusion】 The two matrices for potency detection show good performance including specificity, linearity, repeatability, accuracy, and intermediate precision. They are suitable for the determination of potency in rFⅧ products.

Microsurgical repair of soft tissue necrosis after beak-type calcaneal fracture in 8 cases / 中华显微外科杂志

Objective To investigate the clinical efficacy of microsurgical repair of soft tissue necrosis after beak-type calcaneal fracture.Methods From January,2012 to March,2017,surgically flaps were used to repair wounds in 8 patients with soft tissue necrosis after calcaneal beak fracture.Five patients underwent sural neurovascular flap in the first stage of repair,2 patients were treated with peroneal perforator propeller flap,and 1 patient was treated with posterior tibial artery perforator propeller flap.The donor sites of 3 flaps were directly closed,and donor areas of the remaining 5 were covered with medium-thickness skin grafts without being sutured directly.The size of flap was 5.0 cm× 3.0 cm-7.0 cm × 5.0 cm.Through postoperative outpatient and WeChat follow-up.The patient's flap survival,infection,flap shape,sensation and ankle function were evaluated.Results All flaps and skin grafts survived post-operatively.All patients were followed-up for 6-12 (mean,8.4) months.All patients had good flap survival and no complications such as soft tissue and calcaneal infection.The flaps were good in texture,shape and function of ankle.At the last follow-up,according to the British Medical Research Institute (BMRI),the sensory function was divided into 6 levels.The flap sensory function recovered to S2 in 3 cases,and the remaining 5 cases was S1.According to the American Orthopedic Foot and Ankle Society (AOFAS) Ankle-hindfoot Scale (AHS),the results were excellent in 5 cases,and good in 3 cases.All patients had good clinical results and satisfaction at the last followedup.Conclusion The treatment of soft tissue necrosis after calcaneus beak fractures can be completed in one stage by using flaps,which avoided the occurrence of calcaneal osteomyelitis.It is easy to perform early rehabilitation exercise and the ankle function is well restored.

Surgical treatment of calcaneal fracture malunion / 中华创伤骨科杂志

Objective@#To evaluate the surgical treatment of calcaneal fracture malunion.@*Methods@#A retrospective analysis was conducted of the 19 patients with calcaneal fracture malunion (19 feet) who had been treated from January 2011 to September 2017 at Departments of Joint Surgery and Orthopedic Trauma, The Third Affiliated Hospital, Sun Yat-Sen University. They were 16 males and 3 females with the average age of 36.2 years (from 22 to 57 years). According to Stephens-Sanders classification, the malunion was defined as type Ⅰ in 10 cases, as type Ⅱ in 6 and as type Ⅲ in 3. The patients with malunion of type Ⅰ were treated by lateral wall osteotomy and decompression of long and short peroneus muscles to preserve the subtalar joint, those with malunion of type Ⅱ by lateral wall osteotomy and decompression of long and short peroneus muscles followed by subtalar joint fusion, and those with malunion of type Ⅲ by lateral wall osteotomy, decompression of long and short peroneus muscles, medial oblique osteotomy for correction of varus deformity and subtalar joint fusion via both the medial and lateral approaches. The Maryland functional scores were used to assess the postoperative surgical efficacy.@*Results@#No such complications happened as incision or implant infection, screw breakage or joint non-fusion. Of the 19 patients, 17 were followed up for 18 to 26 months (mean, 20.5 months). The Maryland scores at the final follow-ups were 90.2±7.3, significantly higher than the preoperative values (38.6±5.5) (t=53.370, P<0.001).@*Conclusions@#In the surgical treatment of calcaneal fracture malunion, satisfactory clinical efficacy can be achieved by lateral wall osteotomy and preservation of the subtalar joint for patients with maunion of Stephens-Sanders type Ⅰ, and by subtalar fusion for those with maunion of Stephens-Sanders types Ⅱ-Ⅲ.

Surgical treatment of calcaneal fracture malunion / 中华创伤骨科杂志

Objective To evaluate the surgical treatment of calcaneal fracture malunion.Methods A retrospective analysis was conducted of the 19 patients with calcaneal fracture malunion (19 feet) who had been treated from January 2011 to September 2017 at Departments of Joint Surgery and Orthopedic Trauma,The Third Affiliated Hospital,Sun Yat-Sen University.They were 16 males and 3 females with the average age of 36.2 years (from 22 to 57 years).According to Stephens-Sanders classification,the malunion was defined as type Ⅰ in 10 cases,as type Ⅱ in 6 and as type Ⅲ in 3.The patients with malunion of type Ⅰ were treated by lateral wall osteotomy and decompression of long and short peroneus muscles to preserve the subtalar joint,those with malunion of type Ⅱ by lateral wall osteotomy and decompression of long and short peroneus muscles followed by subtalar joint fusion,and those with malunion of type Ⅲ by lateral wall osteotomy,decompression of long and short peroneus muscles,medial oblique osteotomy for correction of varus deformity and subtalar joint fusion via both the medial and lateral approaches.The Maryland functional scores were used to assess the postoperative surgical efficacy.Results No such complications happened as incision or implant infection,screw breakage or joint non-fusion.Of the 19 patients,17 were followed up for 18 to 26 months (mean,20.5 months).The Maryland scores at the final follow-ups were 90.2 ± 7.3,significantly higher than the preoperative values (38.6 ± 5.5) (t =53.370,P ＜ 0.001).Conclusions In the surgical treatment of calcaneal fracture malunion,satisfactory clinical efficacy can be achieved by lateral wall osteotomy and preservation of the subtalar joint for patients with maunion of Stephens-Sanders type Ⅰ,and by subtalar fusion for those with maunion of Stephens-Sanders types Ⅱ-Ⅲ.

Bundles of care prevents postoperative delirium in elderly patients with hip fractures / 中华创伤杂志

Objective To evaluate the effect of bundles of care on the prevention of postoperative delirium among elderly patients with hip fractures.Methods A prospective randomized case control study was conducted on 80 patients (≥65 years old) with hip fractures from March 2017 to June 2017.The patients were divided into experimental group (n =43) and control group (n =37) according to the random number table method.The experimental group received bundles of care,while the control group received routine nursing.The patients in both groups were all surgically treated,and the confusion assessment method (CAM) was applied to diagnose delirium after surgery.Gender,age,fracture type,duration from injury to operation,internal fixation type,operation time,total amount of bleeding,visual analogue scale (VAS),incidence of delirium,complications,and adverse events were compared between the two groups.Results Among the 80 patients,there were 11 males and 69 females,aged averagely 79.3 years (range,65-95 years).No significant differences were found between experiment group and control group in terms of gender (male:6 cases vs.5 cases,females:37 cases vs.32 cases),age [(79.8 ± 7.8) years vs.(78.7 ± 8.9) years],cause of injury (traffic injuries:7 cases vs.4 cases;falling injuries:36 cases vs.33 cases),duration from injury to operation [(66.1 ±14.3)hours vs.(63.4 ±14.9) hours],fracture type (femoral neck:13 cases vs.10 cases;intertrochanteric:26 cases vs.24 cases;subtrochanteric:4 cases vs.3 cases),internal fixation type (artificial total hip:5 cases vs.5 cases;artificial femoral head:8 cases vs.5 cases;PFNA:29 cases vs.27 cases),operation time [(55.5 ± 16.8) minutes vs.(51.6 ± 17.0) minutes],total blood loss [(114.4 ± 73.9) ml vs.(108.1 ±72.0) ml] (P ＞ 0.05).After bundles of care intervention,the postoperative VAS [(2.2 ± 0.8) points vs.(4.3 ± 1.2) points],postoperative delirium incidence (9％ vs.32％),incidence of complications and adverse events (2％ vs.19％) in experimental group were significantly lower than those in control group (P ＜ 0.05).Conclusion Bundles of care can relieve the pain and effectively reduce the incidences of postoperative delirium,complications,and adverse events in elderly patients with hip fracture.

The effect of percutaneous kyphoplasty combined with Xianling Gubao capsule in treatment of osteoporotic vertebral compression fractures / 中华内分泌外科杂志

Objective To explore the effect of percutaneous kyphoplasty (PKP) combined with Xianling Gubao capsule on treatment of osteoporotic vertebral compression fractures(OVCF).Methods 118 postmenopausal women diagnosed as OVCF and meeting the standard were collected from Mar.2012 to Jul.2016 in our hospital.They were randomly divided into two groups:observation group and the control group.General information of the two groups were comparable.The patients of the control group received PKP combined with traditional anti-osteoporosis therapy.The patients of observation group received Xianling Gubao capsule on the basis of the control group.The lumbar bone density (BMD,greater trochanter BMD,femoral neck BMD,Ward triangle BMD,the Japanese orthopaedic association(JOA)score,the visual analogue score (VAS) score,the serum levels of osteoprotegerin (OPG),receptor activator of nuclear factor-κB ligand (RANKL)and adverse reactions were recorded during the treatment.Results The lumbar BMD,greater trochanter BMD,femoral neck BMD,Ward triangle BMD of observation group at 6-month after operation were higher than those before operation and those of control group (P＜0.05).The VAS score and JOA score of the observation group were superior to those of the control group at 6-month after operation (P＜0.05).The serum levels of OPG of observation group were higher than those of control group,while the serum levels of RANKL of observation group were lower than those of control group at 6-month after operation (P＜0.05).No severe adverse reactions occurred in the two groups during the treatment.Conclusion Xianling Gubao capsule can significantly improve bone metabolism of postmenopausal women with osteoporotic vertebral compression fractures after percutaneous kyphoplasty,increasing BMD,improving lumbar function,relieving pain,without increasing complication rates.

Lengthening the columella of severe secondary bilateral cleft lip nasal deformity by using skin cartilaginous tissue flaps on superior border of bilateral nostril / 中华医学美学美容杂志

Objective To introduce the method of lengthening the columella of severe secondary bilateral cleft lip nasal deformity by using skin cartilaginous tissue flaps on the superior border of bilateral nostril.Methods We adopted the method of using the skin cartilaginous tissue flaps on the superior border of bilateral nostril associated with alar cartilages reduction to lengthen the columella in 40 patients with severe secondary bilateral cleft lip nasal deformity.All patients were followed up for 5-28 months,with an average period of 15.3 months.The treatment outcomes were evaluated by a questionnaire in postoperative follow-up.Results All the 40 patients had achieved satisfactory lengthening of columella and there were no major complications.The questionnaire results showed 36 patients with excellent and 4 patients with good.Conclusions Lengthening the columella of severe secondary bilateral cleft lip nasal deformity by using skin cartilaginous tissue flaps on the superior border of bilateral nostril is a relatively simple and safe method with good effect.

Construction and functional study of three plasmids including Smad3 WT, Smad3 EPSM and Smad3 3S-A stably transfected HepG2 cell lines / 中国药理学通报

Aim Toconstructthreeplasmidsincluding Smad3 WT,Smad3 EPSM and Smad3 3S-A stable transfection in HepG2 cell lines to investigate phospho-domains of Snad3(pSmad3C or pSmad3L),their pro-tein expression and roles in HepG2 cell proliferation, apoptosisandcellcycle.Methods Threeplasmidsin-cluding Smad3 WT (Carry the wild Smad3 gene ), Smad3 EPSM(Carry the mutated phosphorylation site in linker region of Smad3 gene)and Smad3 3S-A(Car-ry the mutated phosphorylation site in C-terminal of Smad3 gene)were respectively transfected into HepG2 cells by using a liposome transfection reagent.Verifi-cation of positive cells was done by screening with G418 via co-culture.Transfection efficiency was deter-mined by Western blot.Cell proliferation was induced by exogenous TGF-β1 in the respective stably transfect-ed HepG2 cell lines.Cell proliferation was monitored by MTT.Cell cycle and apoptosis were determined by flowcytometry(FCM).Results Therewaselevated protein expression of the respective phospho-domain sites in the stably transfected HepG2 cells for Smad3 WT(C-terminus and Linker),Smad3 EPSM(C-termi-nus)and Smad3 3S-A(Linker),which indicated suc-cessful stable transfection of HepG2 cell lines.The re-sults from MTT experiment showed that TGF-β1 could induce proliferation of HepG2 cells with or without the transfection of Smad3 WT,Smad3 EPSM and Smad3 3S-A plasmids,meanwhile transfected Smad3 EPSM plasmids could significantly inhibit proliferation of HepG2 cells induced by TGF-β1 , and transfected Smad3 3S-A plasmids accelerate proliferation of HepG2 cells induced by TGF-β1 .Cell cycle analysis showed that the G0/G1 phase of HepG2 cells with stable trans-fection of Smad3 EPSM plasmid increased compared with HepG2 cells with or without stable transfection of Smad3 WT plasmid,meanwhile the G2/M phase of HepG2 cells with stable transfection of Smad3 3 S-A plasmid increased.Compared with Smad3 WT trans-fected cells, apoptosis in Smad3 EPSM transfected cells was markedly increased,while that of Smad3 3S-Atransfectedcellsdecreased.Conclusions Thethree plasmids of Smad3 WT,Smad3 EPSM and Smad3 3S-A stably transfected HepG2 cell lines have been suc-cessively constructed.The construction of three plas-mids transfected HepG2 cell lines provides the research foundation for studying medical as well as possible reg-ulatory mechanism of pSmad3 C/pSmad3 L.

Nasolabial muscle finite-element study and clinical application / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the nasolabial muscle anatomy and biomechanical characteristics.</p><p><b>METHODS</b>Micro-computed tomography scan was performed in 8 cases of spontaneous abortion fetus lip nasal specimens to construct a three-dimensional model. The nasolabial muscle structure was analyzed using Mimics software. The three-dimensional configuration model of nasolabial muscle was established based on local anatomy and tissue section, and compared with tissue section. Three dimensional finite element analysis was performed on lip nasal muscle related biomechanics and surface deformation in Application verification was carried out in 263 cases of microform cleft lip surgery.</p><p><b>RESULTS</b>There was close relationship between nasolabial muscle. The nasolabial muscle tension system was constituted, based on which a new cleft lip repair surgery was designed and satisfied results were achieved.</p><p><b>CONCLUSIONS</b>There is close relationship among nasolabial muscle in anatomy, histology and biomechanics. To obtain better effect, cleft lip repair should be performed on the basis of recovering muscle tension system.</p>

Suckling mouse model establishment of myocarditis induced by foot-and-mouth disease vi rus / 中国人兽共患病学报

Interactions between FMDV and cardiac cells are multifaceted and complex ,these interactions leads to pro-teins alterations in cardiac cells inevitably .To understand the pathogenesis of myocarditis after FMDV infection in mice ,the suckling mouse model for myocarditis induced by foot-and-mouth disease virus (FMDV) was established in this study .Suckling mice within 3 days old was selected to infect by FMDV .Myocarditis caused by FMDV in suckling mice was confirmed with clinical symptom monitor .The observation of Hematoxylin and eosin stain (H&E stain) and transmission electron microscopy (TEM) were performed after samples processing .According to conventional polymerase chain reaction (PCR) methods ,prim-ers of VP1 gene was designed ,synthesised and specific FMDV VP1 gene was amplified from the heart muscle of suckling mice . The results indicated that suckling mice appeared low spirit condition ,dyspnea ,and dull reaction within 36 hours after chal-lenge with FMDV .Infiltration of inflammatory cells and dissolution of myocardial fibers were observed with H&E stain and TEM .Special target gene of FMDV was amplified from the heart of infected group .Obvious inflammation in the heart of suck-ling mice caused by FMDV was observed .It's suggested that suckling mouse model for myocarditis induced by FMDV was es-tablished successfully ,which would lay the foundation for researches of myocarditis mechanism in young cloven-hoofed ani-mals .

An unique exonic splicing mutation in the CYPI7AI gene as the cause of steroid 17α-hydroxylase deficiency / 中华内分泌代谢杂志

Objective To analyze CYP17A1 gene mutation in a patient with 46,XY disordered sex development and to explore the possible influence on the phenotype of the patient.Methods Eight exons of CYP17AI gene in the patient and her parents were amplified and directly sequenced.In order to construct Mini-gene system,PCR fragments containing wildtype and mutant splicing sites were inserted in expression vector,and then transfected into cells.RT-PCR was used to observe the influence of splicing site mutation.Wildtype and aberrant splicing CYP17A1 cDNA expression plasmids were constructed and transfected into cells respectively,and CYP17A1 enzyme activity was tested in vitro.Results Mutation analysis revealed compound heterozygous CYP17A1 mutations,with Y329fs in one allele and a synonymous substitution( c.1263G＞A:GCG＞GCA) in another allele.In vitro analysis showed that the synonymous substitution induced a novel splicing site,which resulted in aberrant splicing of CYP17A1 mRNA and lacked six or seven amino acids after 415 in splicing product.In vitro transfection and enzyme activity experiment showed that the aberrant splicing product abolished the enzyme activity completely.However,this mutation did not completely influence splicing.The patient also had a part of normal splicing product,which was a coincidence to the phenotype of the patient.Conclusion This is the first description of an exonic splicing mutation in CYP17A1 relevant to the 17ot-hydroxylase deficiency phenotype.The functional study of the aberrant splicing variant has been initiated.

Genetic diagnosis and weight loss surgery of a case of Prader-Willi syndrome / 中华内分泌代谢杂志

To investigate the clinical features, genetic diagnosis, and treatment of a patient with Prader-Willi syndrome(PWS). For a case with clinically suspected PWS, methylation specific PCR(MSPCR)amplification was applied to CpG islands of SNRPN(exon α)gene locus in the 15q11-q13. Furthermore, the diagnosis was comfirmed by the method of bisulfite sequencing PCR(BSPCR). Metabolic status before and after the operation of sleeve gastrectomy were compared. Absence of amplification of paternal allele on chromosome 15q11-q13 was detected in the case by MSPCR, different from the normal control. Results of BSPCR further proved a full methylation of CpG islands in the SNRPN gene locus. Four months after sleeve gastrectomy, systemic metabolic status and ventricular function were improved. MSPCR and BSPCR were both consistent with genetic diagnosis of PWS. Weight loss surgery is expected to be a major therapy of this disease.

Study of transdiaphragmatic pressure and its correlation with esophageal pressure in ARDS piglet / 中华急诊医学杂志

Objective To study the changes of trans-diaphragmatic pressure (Ptra) and its correlation with esophageal pressure (Peso) through ARDS piglet model.Methods Five piglets were enrolled in the study.Peso,gastric pressure (Pgas) and intra-thoracic pressure (Pint) was monitored through balloon inserted.The data before ARDS serve as control.ARDS was produced in the piglets through saline lavage.The pressure were observed and the Ptra were calculated.The pressure changes and correlation between Ptra and Peso were analyzed as well.Linear regression with the coefficient of determination and t-test were used as appropriate.Significance was assumed for P ＜ 0.05.Results Peso,Pgas and Pint before ARDS were 7.3 ± 1.9,25.5 ± 2.4,- 1.23 ± 0.21 cmH2O,Ptra was 18.2 ± 1.6 cmH2O.While after ARDS,the data were 4.7 ± 1.4,31.1 ± 3.1 and - 1.79 ± 0.28 cmH2O,and Ptra was 26.4 ± 2.1 cmH2 O,and all these changes were obviously ( P ＜ 0.05 ).The correlation between Pint and Peso,Pint and Ptra (A) and Ptra ( B ) were 0.93 ± 0.025,0.88 ± 0.023 and 0.87 ± 0.37 before ARDS.After ARDS,the correlation changed to be 0.82 ±0.21,0.81 ±0.20 and 0.78 ±0.31.Although a bit decreased,the correlation was still positive (P ＜ 0.01 ).Conclusions There existed good correlations between Peso and Ptra as well as between Pint and Peso before or after ARDS.Ptra was increased obviously after ARDS,which could lead to respiratory muscle fatigue.

Expression of goose interleukin-2 gene in Escherichia coli and isolation of its soluble monomer / 生物工程学报

Recombinant expression plasmid of pET-28a (+)-goIL-2 was constructed by inserting the goose IL-2 gene without the signal peptide sequence into the prokaryotic expression vector pET-28a (+), and transformed into the bacterial competent E. coli BL21 (DE3) cells for expression. After IPTG induction, an expected protein band with molecular weight of 15.0 kD was observed on SDS-PAGE gel, recognized by monoclonal antibody against goose IL-2 in western-blotting assay. In the pET-28a (+) expression system, much of the recombinant goose IL-2 (rgoIL-2) was found in inclusion bodies with a portion of soluble protein. The monomer and multimers of soluble goose interleukin 2 proteins were observed in native electrophoresis. The rgoIL-2 proteins were purified by Ni-NTA column under a native condition. The rgoIL-2 soluble protein monomer was isolated by a quick protein isolation and purification system of AKTA FPLC and identified by native PAGE. Bioactivity analysis showed that the rgoIL-2 monomer stimulated the proliferation of goose lymphocytes in vitro. This will establish a basis for further study about the biological function and clinical application of goose IL-2.

Clinical significance of histological examination and noninvasive measurements in renal osteodystrophy / 中华肾脏病杂志

Objective To analyze the histological changes of bone diseases and to investigate the noninvasive measurements for diagnosing renal osteodystrophy (ROD) in maintenance dialysis patients . Methods Ninety-one patients were selected to receive bone biopsy . The bone samples were stained with HE, toluidine blue and Masson, and were examined with light microscopy . The levels of immunoreactive parathyroid hormone (iPTH), osteoprotegerin (OPG),sRANKL and osteocalcin (OCN) were determined in the patients enrolled from 2004 to 2006 . The level of iPTH was measured by radioimmunoassay . OPG and sRANKL were measured by ELISA,and OCN was measured by chemiluminescence . Results The incidence of ROD in the maintenance patients was 100% . According to the histological appearance, 50 cases (54 .9%) were high turnover bone disease (secondary hyperparathyroid bone disease), 9 cases (9 .9%) were low turnover bone diseases(osteomalacia and adynamic bone disease), and 32 cases(35 .2% ) were mixed bone disease . The level of iPTH in patients with ROD was significantly increased compared with healthy controls . It was the lowest in low turnover bone diseases . There was no difference among three types of ROD . OPG level was significantly increased compared with healthy controls [(2176 .58±1576 .08) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . The level in high turnover bone diseases was higher than that of the healthy controls [(2261 .85±1712 .22) pmol/L vs (1310 .46±1254 .00) pmol/L, P＜0 .05] . There was no difference among three types of ROD .sRANKL level in high turnover bone disease was significantly increased compared with healthy controls [(0 .328±0 .524)pmol/L vs (0 .084±0 .190) pmol/L, P＜0 .05] . OCN level was also higher than that of the healthy controls (P＜0 .05), and the OCN level in low turnover ROD was the lowest among three types of ROD . OCN level in mixed ROD was dramatically increased as compared to low turnover ROD [(226 .633±66 .455) pmol/L vs (193 .03±104 .269) pmol/L, P ＜0 .05] .Conclusions The histological changes of bone disease can be indicated by iPTH level, but the types of ROD can not be distinguished according to iPTH level neither be differentiated by the levels of OPG, sRANKL and OCN . Bone histomorphometry is still the golden standard for diagnosing renal osteodystrophy .