Objective To investigate the role of Twist and vascular endothelial growth factor (VEGF)in the development of epithelial ovarian cancer and their relationship with clinical pathological features.Methods The expressions of Twist and VEGF were detected by immunohistochemical staining (SP)method in 80 cases of epithelial ovarian cancer and 10 paired cases of normal ovarian tissue.Results In the 80 cases of ovarian cancer tissues,the positive expression rate of Twist and VEGF in epithelial ovarian cancer was 85% and 86.25%,respectively,which were remarkably higher than those in normal ovarian tissues (P <0.01).The expressions of Twist and VEGF were closely associated with clinical stages,pathological grades and lymph node metastasis,but not correlated with age or pathological patterns (P >0.05).There was a significant correlation between the expressions of Twist and VEGF in epithelial ovarian cancer (r =0.646,P <0.01).Conclusion The high expressions of Twist and VEGF may play a potential role in the occurrence,development and invasion process of epithelial ovarian cancer.

Objective To approach the neurobiochemical mechanism of chronic central pain (CCP) after spinal cord injury (SCI). Methods 28 SD rats were divided into four groups, the normal group (group A), the pseudosurgery group (group B), and groups with CCP (group C) and without CCP (group D) after L1 spinal cord section injured with WADE method. T13 and L2 segments of rats' spinal cord were took and concentration changes of substance P (SP) in the spinal dorsal horn between two sections were examined by immunofluorescence histochemistry staining combined with confocal laser scanning microscope. Results Concentration of SP in the group D was decreased significantly compared with groups C,A and B (P<0.05-0.01), that of the group C was less than that of group A and B (P<0.05). Conclusion The rat model established by WADE method is proper to study CCP after SCI. SP in dorsal horn of spinal cord may inhibit the CCP after SCI in some degrees.

Objective:To investigate the changes of blood pressure and heart rate in patients aged over 80 years undergoing intravitreal injection of anti-vascular endothelial growth factor(VEGF)drugs under topical anesthesia without anesthetic monitoring in ophthalmology surgery, and to analyze the potential risk for surgery-induced acute cardiovascular and cerebrovascular events.Methods:This was a prospective observational cohort study.A total of 100 patients(aged 80 to 95 years, 47 males and 53 females)included 85 cases of wet age-related macular degeneration, 14 cases of retinal vein occlusion with macular edema, and 1 case of diabetic macular edema.These patients received anti-VEGF drugs injection, with accumulated injections of 1 to 36 times per patient.Blood pressure and heart rate were measured at visit to a medical clinic(as baseline), before operation, 0 min and 30 min after operation.The operation anxiety and pain scores were investigated by questionnaires.Comparison of systolic blood pressure, diastolic blood pressure and heart rate among four different time points was performed by Wilcoxon symbol rank test.Results:At four different time points i. e.at the baseline, before injection, 0 min and 30 min after injection, systolic blood pressure, diastolic blood pressure and heart rate showed an important regularity.The median systolic blood pressure at the four time points was 130(120-140)mmHg(1 mmHg=0.133 kPa), 142(129-149)mmHg, 153(139-164)mmHg and 143(130-151)mmHg, with ranges of 100-160 mmHg, 106-176 mmHg, 118-197 mmHg and 115-187 mmHg, respectively.The median diastolic blood pressure at the four time points was 70(65-75)mmHg, 76(69-83)mmHg, 81(73-87)mmHg and 75(69~81)mmHg, with ranges of 45-90 mmHg, 55-98 mmHg, 58-99 mmHg and 50-93 mmHg, respectively.The median heart rate at the four time-points was 70(65-80)beats/min, 72.5(65-81)beats/min, 73(66-80)beats/min and 70(65-76)times/min, with ranges of 50-95 beats/min, 48-101 beats/min, 51-93 beats/min and 50-87 beats/min, respectively.Systolic blood pressure and diastolic blood pressure were significantly increased from preoperative time to 0 min and 30 min after operation( P<0.01). Compared to the baseline, heart rate was increased before operation( P<0.01)and was decreased at 30 minutes after operation( P<0.01). Conclusions:Blood pressure fluctuation may exceed the safe range with an obvious increase of systolic blood pressure during the current intravitreal injection of anti-VEGF drugs in the patients aged over 80 years.Systolic blood pressure rises significantly before operation and falls back after operation, but it may last till 30 minutes after operation.The range of heart rate change is relatively small.Perioperative management and monitoring measures should be strengthened to reduce the risk of acute cardiovascular and cerebrovascular events.

To obtain single chain variable fragment (scFv) and bivalent single chain variable fragment (bsFv) against transferrin receptor, up-stream and down-stream primers were designed according to the complementary sequences of FR1 region of variable heavy (VH) and FR4 of variable light (VL), respectively, which contained inter-linker G4S and the restriction endonuclease SfiI, AscI and NotI. Two pieces of scFv fragments were first amplified through PCR and then inserted into plasmid pAB1, which could express scFv protein once induced by IPTG in the host bacteria. To express scFv and bsFv, E. coli TG1 was cultured in LB broth and was induced by IPTG. The restriction enzyme digestion map and DNA sequencing demonstrated that scFv and bsFv genes were successfully inserted into the expression plasmid. SDS-PAGE and Western blotting revealed the protein band at 35kD and 60kD, which were consistent with the molecular weight of scFv and bsFv respectively. Flow cytometry showed that scFv and bsFv harbored the specific binding activity with TfR expressed in various tumor cells, and the avidity of bsFv was higher than that of the parent scFv.
Base Sequence ; Cloning, Molecular ; Escherichia coli/genetics ; Escherichia coli/metabolism ; Genetic Vectors/genetics ; Hep G2 Cells ; K562 Cells ; Molecular Sequence Data ; Receptors, Transferrin/*immunology ; Recombinant Fusion Proteins/biosynthesis ; Recombinant Fusion Proteins/genetics ; Single-Chain Antibodies/*biosynthesis ; Single-Chain Antibodies/genetics

OBJECTIVETo investigate the clinical outcomes of the posterior C1,2 screw-rod combined with C2 unilateral translaminar screw and contralateral pedicle screw fixation and autogenous bicortical iliac crest graft fusion in treating upper cervical instability with vertebral artery variations.

METHODSFrom June 2008 to December 2012, the clinical data of 12 patients with upper cervical instability underwent C1 lateral mass screws-C2 unilateral laminar and contralateral pedicle screws fixation combined with autogenous bicortical iliac crest graft fusion were analyzed retrospectively. There were 8 males and 4 females with a mean age of 47.5 years (ranged, 16 to 77 years). Patients suffered from occipitocervical activity limitation of motion with pain or not, VAS was 0-7 points with an average of (3.50 +/- 2.71) points. Unilateral vertebral artery hypoplasia was demonstrated by vertebral arteriography (VAG) or CTA in all patients. Cervical X-ray and CT scans were done within 7 days after surgery in order to confirm internal fixation position. Internal fixation loosening and breakage, reduction losing, bone fusion ratio were observed during follow-up.

RESULTSNo nerves and vertebral artery injuries occurred during operation. Cervical pain obviously decreased and VAS was (0.92 +/- 0.90) points. Cervical alignment of 12 patients had well-recovered by X-ray while Atlantoaxial ventral lamina cortex of 1 case was encroached by CT scan without neurological symptom. All patients were followed up for 6 months to 3 years, no internal fixation loosening and breakage, reduction losing were found. All patients obtained bone fusion in 6-12 months after operation.

CONCLUSIONPosterior C1 lateral mass screws-C2 unilateral laminar and contralateral pedicle screws fixation combined with autogenous bicortical iliac crest graft fusion can achieve biomechanical stability and raise the successful rate of bone fusion, while avoiding the risk of vertebral artery injury and overcoming the insufficient of bone fusion during bilateral laminar screws placement as well. Posterior C1 lateral mass screws fixation is a safe and effective additional method in treating upper cervical instability with vertebral artery variations.

Adolescent ; Adult ; Aged ; Bone Screws ; Cervical Vertebrae ; surgery ; Female ; Humans ; Internal Fixators ; Joint Instability ; surgery ; Male ; Middle Aged ; Tomography, X-Ray Computed ; Vertebral Artery ; pathology

AIM:To explore the application of 10g/L cyclopentolate chloride eye drops in children, and to compare the different effectiveness of cycloplegia between 10g/L cyclopentolate chloride and atropine in Chinese children.METHODS:A total of 236 eyes of 118 children aged 3~12 years old were enrolled in this study including 80 eyes of 40 children with myopia, 156 eyes of 78 children with hyperopia and 146 eyes of 73 children combined with astigmatism. 10g/L cyclopentolate chloride eye drops were used once per 5min for 3 times and refractive diopter was obtained 1h after the last drop of cyclopentolate. Three days after that, 10g/L atropine was then used 1 time per night for 1wk and optometry was performed again. The children were divided into 3 groups ( myopia, hyperopia and astigmatism group ) according to the refractive status, in which astigmatism was independent of the degree of separation of cylinder statistics. The results of retinoscope refraction were then compared between 10g/L cyclopentolate and 10g/L atropine.
RESULTS:The refractive diopter was -2. 25±1. 31D after 10g/L cyclopentolate eye drops and -2. 23±1. 32D after 10g/L atropine in myopic group. The refractive diopter was 1. 35±1. 19D and 1. 38±2. 00D in astigmastic group. No significant difference was found in myopic group and astigmastic group (P= 0. 109, P= 0. 374). While in the hyperopic group, the refractive diopter was 3. 76±2. 4D after 10g/L cyclopentolate eye drops, which was lower than that after 10g/L atropine 4. 39±2. 6D (P=0. 000).
CONCLUSION: The results of this study suggest that 10g/L cyclopentolate chloride eye drops can be used in myopia and astigmatism children, and 10g/L atropine should be used in hyperopia children.

OBJECTIVETo explore the correlation among prevertebral hyperintensity (PVH), sagittal canal diameter on MRI and neurologic function of patients after cervical vertebral hyperextension injury without fracture and dislocation.

METHODSThe clinical data of 100 patients with cervical vertebral hyperextension injury without fracture and dislocation were retrospectively analyzed from September 2010 to December 2013. The patients were divided into PVH group and non-PVH group according to the presence of PVH on T2-weighted magnetic resonance imaging. There were 39 patients in PVH group, including 31 males and 8 females, aged from 21 to 83 years old with an average of (58.10 ± 14.78) years; and the other 69 patients in non-PVH group, including 49 males and 12 females, aged from 32 to 77 years old with an average of (55.05 ± 10.36) years. The sagittal disc level canal diameters of subaxial cervical spine were measured on mid-sagittal magnetic resonance imaging. The age, sex, cause of injury, and the segments of spinal stenosis were recorded. American Spinal Injury Association (ASIA) impairment scale and motor score were used to evaluate the neurological status.

RESULTSThe ASIA motor score of the group with PVH was 52.56 ± 31.97 while the ASIA motor score was 67.70 ± 22.83 in non-PVH group (P = 0.013). More patients with intramedullary hyperintensity signal on MRI were observed in the PVH group than in non-PVH group (P = 0.006). There was a significant positive correlation between ASIA motor score and sagittal disc level canal diameter of injury segment (P = 0.003). The neurological status was worse in patients with multi-level sagittal canal diameters below 8 mm.

CONCLUSIONThe PVH and the disc-level canal sagittal diameter of the injury segment are associated with neurological status. The patients with multi-level sagittal canal stenosis are vulnerable to severe cervical spinal cord injury.

Adult ; Aged ; Aged, 80 and over ; Cervical Vertebrae ; injuries ; Female ; Humans ; Magnetic Resonance Imaging ; Male ; Middle Aged ; Retrospective Studies ; Spinal Canal ; pathology ; Spinal Cord Injuries ; pathology ; physiopathology

Objective To investigate the role of the p38 MAPK pathway in the formation of cytoplasmic vacuoles .Methods Af-ter treated with Anisomycin ,SB203580 or SP600125 ,images of HepG2 ,LM3 ,QBC939 ,Hela and A549 cells were recorded by light microscopy and taken at a magnification of 400 × .The effects of anisomycin ,SB203580 and SP600125 on the activity of p38 and JNK were measured by Western blot .LM3 and A549 cells were stained with the ER-tracker red and the lyso-tracker red and subjec-ted to confocal microscopy analysis .Results (1)Anisomycin could abolish cytoplasmic vacuolization of HepG2 cells .(2)p38 MAPK activation was responsible for anisomycin-induced cytoplasmic vacuolization abolishment .(3)p38 MAPK blocking initiated cytoplas-mic vacuoles formation in various cancer cell lines .(4)p38 MAPK blocking-induced cytoplasmic vacuoles disrupted the integrity of endoplasmic reticulum .(5)p38 MAPK blocking reversibly induced cytoplasmic vacuoles formation .Conclusion These observations provide direct evidence for a role of p38 MAPK signaling in regulating the formation of cytoplasmic vacuoles .

Objective To construct vector expressing soluble mPDL1-hIgGFc and study its effect on the proliferation and apeptosis of cells in vitro. Methods The extrncellular domain of mPDL1 gene was amplified from pmPDL1 vector by PCR and inserted into phIgGFc vector. The recombinant pmPDL1-hIgGFc was transfected into CHO cells by LipofectAMINETM2000, and the transfected cells were named as CHOp. The expression of mPDL1-hIgGFc in the culture supernatants of CHOp was assayed by ELISA and Western blot. The effects of CHOp culture supernatants on mixed lymphocyte culture(MLC) was analysed by Flowm-etry. Results The extracellular domain of mPDL1 gene were obtained from PCR. DNA sequencing and the identification of digestion by HindⅢ and KpnⅠ indicated the recombinant plasmid pmPDL1-hIgGFc was suc-cessfully constructed. ELISA and Western blot analysis proved that the CHOp could express mPDL1-hIgG-Fc. CHOp culture supernatants could inhibit lymphocyte proliferation and induce the apoptosis of the activa-ted T cells in MLC in vitro in a dose-dependent manner. Conclusion The mPDL1-hIgGFc protein could in-hibit lymphocyte proliferation and induce the apoptosis of the activated T cells.

Objective To investigate the intelligence standard for diagnose the sub-cretin children and children with mental retardation of socio-cultural type.Methods The full intelligence quotient(IQ),verbal intelligence quotient(VIQ)and performance intelligence quotient(PIQ)was tested by Wechsler scale(C-WISC)for mild iodine deficiency disordem children,children living in abnormal socio-cultural condition and normal children aged 7～14 years old in Qinba mountain area.The test results had been compared between the groups.Results There were no significant difference between psychomotor functioning well children and children living normal sociocuhural condition in VIQ,PIQ and full IQ(89.24±18.44 vs 90.75±17.58,87.58±15.78 vs 88.95±15.56,87.42±17.84 vs 89.02±17.18,t=1.14,1.19 and 1.24,respectively,all P＞O.05).PIQ and full IQ were significantly lower in mild iodine deficiency disorders children than in children with abnormal socio-cultural background (65.81±10.22 vs 72.33±13.23,62.42±12.31 vs 68.13±14.54,t=3.26,2.55,P＜0.01 or＜0.05,respectively).But the VIQ was not significantly different between these two groups.The average difference of VIQ and PIQ among mild iodine deficiency disorders children wag-0.32 without significant difierence(t=0.28,P＞0.05),however it was-2.91 among children under abnormal socio-cultural condition with significant difierenee(t=-3.59,P＜0.01).Conclusions IQ for iodine deficiency disorders children is characterized by that VIQ is damaged in parallel with PIQ,while that in children under abnormal soeio-cuhural condition is marked by that VIQ is retarded more severely than PIQ,which ean be used as an intelligence standard for differentiating the sub-cretin children from children wjth socio-cuhural mental retardation.