Objective To develop nanometer-scale bubbles with surfaces of N-palmitoyl chitosan(PLCS) as ultrasound contrast agent and evaluate its characteristics and acoustic effects in vivo. Methods The PLCS nanobubbles were prepared using a cutting technique at differential high-frequency of shear speed. Both optical and transmission electron micrography were performed to determine the nanobubble size and morphology. Concentration, size-distribution and zeta potential of the PLCS nanobubbles were measured by cell counting chamber, Malvern lazer particle analyzer and zeta-sizer at 1-day, 45-day and 90-day. The acoustic effects of the PLCS nanobubbles on myocardium and renal tissue in 6 normal rats were observed using bolus infusion of the nanobubbles intravenously. The maximum video intensity(VI) was measured.Results The PLCS nanobubbles with nice round-shape and uniform site-distribution were demonstrated.The mean diameter,concentration and zeta potential of the PLCS nanobubbles were (617 ± 12) nm, (7.2 ±0.6) × 109/ml and (52.9 ± 1.3)mV at the 1-day,and all of parameters did not change significantly in 45-day and 90-day ( P ＞ 0. 05). A significant contrast-enhancement was noted on myocardium and renal tissue during infusion of the nanobubbles. VI on both tissues was (15.6 ± 1.1)GU and (27.3 ± 2.5)GU,respectively. The visual contrast-enhancement last up to (10 ± 2)min. Conclusions The PLCS nanometerscale bubbles have excellent physical-features and contrast-enhanced ultrasound effects in vivo. It may develop as a novel contrast ultrasound agent which could cross endothelial cell membrances.

BACKGROUND: The correlation of gycosylphosphatidilinoditol-specific phospholipase D (GPI-PLD) activity, mRNA expression to leukemia type, hepatosplenomegaly and/or lymphadenopathy has been rarely reported. OBJECTIVE: To explore the correlation of GPI-PLD expression to leukemia type and hepatosplenomegaly and/or lymphadenopathy of acute myeloid leukemia (AML) patients. METHODS: Fresh bone marrow specimens were obtained from 43 newly diagnosed AML patients, 28 acute lymphocytic leukemia (ALL) patients, and 21 normal persons. Bone marrow mononuclear cells were harvested by density gradient centrifugation. GPI-anchored human placent alkaline phosphatase was used as substrate. GPI-PLD activity was determined bytriton-X114 phase partitioning procedure. GPI-PLD mRNA expression was detected by semi-quantitative RT-PCR. The relationship of GPI-PLD activity, mRNA expression and leukemia type, hepatosplenomegaly and/or lymphadenopathy was analyzed. RESULTS AND CONCLUSION: Compared with control group, GPI-PLD activity and mRNA expression in bone marrow mononuclear cells were significantly higher in AML group (P < 0.01), while they were significantly lower in the ALL group (P < 0.01). Of 43 patients with AML patients, 13 patients had hepatosplenomegaly and/or lymphadenopathy. The GPI-PLD activity (%) and mRNA expression were significantly higher in AML patients without hepatosplenomegaly and lymphadenopathy than those patients with hepatosplenomegaly and/or lymphadenopathy (P < 0.05). These results demonstrated that GPI-PLD activity alteration is consistent with GPI-PLD mRNA expression in AML patients, and the expression levels correlate to leukemia type and hepatosplenomegaly and/or lymphadenopathy of AML patients.

To construct a plasmid expressing MDR1 short hairpin RNA (shRNA) mediated by pEGFP-C1/U6 vector, two coding sequences of 19 nucleotides were selected from MDR. Two pairs of oligonucleotides were designed for these two fragments. After annealing the formed double-stranded DNAs were ligated with plasmid pEGFP-C1/U6 (pEGFP-C1 vector with U6 promoter). The plasmids producing MDR1 shRNA were constructed from the inverted motif containing 9 spacers and four Ts. The results showed that the constructed plasmids were named pEGFP-C1/U6/A and pEGFP-C1/U6/B, and the constructs were identified by restriction and sequence analysis, no any base mutation was observed. It is concluded that plasmids of pEGFP-C1/U6/A and pEGFP-C1/U6/B expressing MDR1 shRNA were successfully constructed, providing a highly effective method for reversing the multidrug resistance in clinic.
ATP-Binding Cassette, Sub-Family B, Member 1 ; biosynthesis ; genetics ; DNA-Binding Proteins ; Drug Resistance, Multiple ; genetics ; Drug Resistance, Neoplasm ; genetics ; Genetic Vectors ; genetics ; Green Fluorescent Proteins ; genetics ; Humans ; Plasmids ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; RNA, Small Interfering ; biosynthesis ; genetics ; RNA, Small Nuclear ; genetics

OBJECTIVETo explore a new experimental method for screening of allergens in post-market traditional Chinese medicine injections by confirming allergens in Huachansu injection.

METHODFirst of all, the serum of patients allergic to Huachansu injection were collected, at the same time, the dubious allergen was conjugated to bovine serum albumin (BSA) by EDC coupling procedure to form complete antigen (BNP-BSA), which makes it possible to reproduce the allergic reaction of Huachansu injection in vitro. The histamine liberation ratio, the level of TNF-alpha and Histamine released from RBL-2H3 mast cell were detected; the above data were compared with those obtained in vivo.

RESULTThe difference of the histamine liberation ratio, the levels of TNF-alpha and histamine of the resibufogenin-BSA group, group of patients allergic to Huachansu injection were not significant compared with those of normal control group. However, there were significant difference in those data among the cinobufagin-BSA group, the blank control and normal control group (P<0.05).

CONCLUSIONThe allergen in the serum collected from patients allergic to Huachansu injection is resibufogenin.

Allergens ; adverse effects ; immunology ; Amphibian Venoms ; adverse effects ; immunology ; Animals ; Anura ; Bufanolides ; adverse effects ; immunology ; Drug Hypersensitivity ; immunology ; Histamine Release ; Humans ; Mast Cells ; immunology ; Medicine, Chinese Traditional

OBJECTIVETo discuss the protective effect of Mailuoning injection on ischemia/reperfusion (I/R) injury in rats and its mechanism.

METHODHealthy male adult Sprague-Dawley (SD) rats were randomly divided into the sham operation group, the model group, the edaravone (3 mg x kg(-1)) control group, and Mailuoning high, middle and low-dose groups (4, 2, 1 mL x kg(-1)), with 10 rats in each group, and administered with drugs through tail intravenous injection. The middle cerebral artery occlusion (MCAO) was adopted to establish the rat ischemia/reperfusion model. After the ischemia for 2 h and reperfusion for 24 h, the pathological changes in neurovascular units (NVU) of brain tissues at the ischemia side was observed by HE staining. The expressions of glialfibrillary acidic protein (GFAP) and ionized calcium-binding adaptor molecule 1 (Ibal) were detected by the immunohistochemical method. The expressions of tumor necrosis factor-alpha (TNF-alpha), interleukin 1beta (IL-1beta), vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) were detected by the western blotting technique.

RESULTMailuoning injection could significantly improve the pathological changes in cortical penumbra brain tissue UVN of (I/R) rats, reduce the number of GFAP and Ibal positive cells, and significantly decrease the expressions of TNF-alpha, IL-1beta, VCAM-1 and ICAM-1 of brain tissues of I/R rats.

CONCLUSIONMailuoning injection shows an obvious protective effect on UVN of I/R rats. Its mechanism may involve the inhibition of the activation of astrocyte and microglia and the secretion and expression of various inflammatory factors.

Animals ; Brain ; drug effects ; metabolism ; Brain Ischemia ; surgery ; Drugs, Chinese Herbal ; administration & dosage ; Humans ; Infarction, Middle Cerebral Artery ; genetics ; metabolism ; Intercellular Adhesion Molecule-1 ; genetics ; metabolism ; Male ; Protective Agents ; administration & dosage ; Rats ; Rats, Sprague-Dawley ; Reperfusion Injury ; genetics ; metabolism ; prevention & control ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; Vascular Cell Adhesion Molecule-1 ; genetics ; metabolism

Bronchopulmonary Sequestration ; classification ; pathology ; surgery ; Humans ; Infant ; Lung ; diagnostic imaging ; Male ; Radiography ; Respiratory System Abnormalities ; diagnosis ; surgery

Objective:To investigate the expression levels of matrix metalloproteinase-9 (MMP-9), procalcitonin (PCT), soluble triggering receptor expressed on myeloid cells (sTREM-1) and soluble cell differentiation 14 (sCD14) in pregnant women with premature rupture of membranes (PROM) and their predictive value for chorioamnionitis.Methods:A total of 132 pregnant women with PROM who received treatment in Tengzhou Central People's Hospital from January 2016 to June 2017 were included in the study group. These women were assigned to pre-term PROM group (gestational age < 37 weeks, n = 58) and full-term PROM group (gestational age > 37 weeks, n = 74). A total of 106 concurrent full-term healthy pregnant women were included in the control group. Pregnant women in the PROM group were also assigned into an infection group ( n = 51) and a non-infection group ( n = 81). Serum levels of MMP-9, PCT, sTREM-1 and sCD14 were compared between study and control groups, and their value in the diagnosis of PROM complicated with chorioamnionitis was analyzed. Results:The expression levels of MMP-9 [(271.42 ± 34.16) ng/L], PCT [(54.57 ± 8.16) pg/mL], sTREM-1 [(0.51 ± 0.11) ng/mL] and sCD14 [(60.23 ± 9.49) ng/mL] in the study group were significantly higher than those in the control group [(54.97 ± 10.08) ng/L, (26.04 ± 1.98) pg/mL, (0.19 ± 0.04) ng/mL, (42.04 ± 10.33) ng/mL, t = 27.064, 13.767, 14.831, -13.342, all P < 0.01). The expression levels of MMP-9 [(314.05 ± 45.37) ng/L], PCT [(0.61 ± 0.18) ng/mL], sTREM-1 [(63.12 ± 10.12) pg/mL] and sCD14 [(68.07 ± 11.05) ng/mL] in the pre-term PROM group were significantly higher than those in the full-term PROM group [(238.01 ± 40.45) ng/L, (47.87 ± 8.90) pg/mL, (0.43 ± 0.14) ng/mL, (54.09 ± 10.33) ng/mL, t = 9.103, 8.862, -10.538, 6.494, all P < 0.05). The expression levels of MMP-9 [(343.74 ± 43.74) ng/L], PCT [(69.88 ± 8.83) pg/mL], sTREM-1 [(0.67 ± 0.16) ng/mL], sCD14 [(70.41 ± 8.89) ng/mL] in the infection group were significantly higher than those in the non-infection group [(230.09 ± 49.82) ng/L, (45.82 ± 11.04) pg/mL, (0.42 ± 0.19) ng/mL and (54.41 ± 12.42) ng/mL, t = 23.655, 12.014, 9.382, 11.306, all P < 0.01]. The sensitivity (94.23%), specificity (93.75%), positive predictive value (92.45%) and negative predictive value (96.20%) of combined detection of these indexes in the diagnosis of PROM complicated by chorioamnionitis were significantly higher than those of other indexes detected alone (all P < 0.05). Conclusion:Combined detection of serum levels of MMP-9, PCT, sTREM-1 and sCD14 can be used as an effective auxiliary index for the diagnosis of early premature rupture of membranes complicated with chorioamnionitis.

AIM:To investigate the surgical method and extent of reoperation in the concomitant strabismus patients with surgical under-correction and over-correction.
METHODS: Ninety - six concomitant strabismus patients with surgical under - correction and over -correction were recruited in this study, which included 41 males and 55 females, aged 21. 90±14. 70. All individuals underwent routine eye examinations for strabismus before the surgery. Among the cases with concomitant esotropia, there were over-correction in 23 cases, under-correction in 15 cases. Among the cases with concomitant exotropia, there were over-correction in 28 cases, under - correction in 30 cases. The method of reoperation were based on angle of deviation, the method of original operation and acute visual acuity of patients.
RESULTS:In over - correction cases with concomitant esotropia, medial rectus muscle of 9 cases were advanced, the corrective extent was (5. 51±2. 63) ? / mm;9 cases were performed advance of medial rectus muscle and recession of lateral rectus muscle, the corrective extent was (6. 25±1. 59) ? / mm; 3 cases were performed resection of medial rectus muscle and recession of lateral rectus muscle, the corrective extent was (4. 26±1. 04) ? /mm; only 2 cases were performed recession of lateral rectus muscle, the corrective extent was (4. 21±1. 91) ? /mm. In under - correction cases with concomitant esotropia, 6 cases were performed resection of lateral rectus muscle, the corrective extent was (4. 03±0. 98) ? /mm; 6 cases were performed resection of lateral rectus muscle and recession of medial rectus muscle, the corrective extent was (6. 86 ± 1. 32) ? / mm; 3 cases were performed recession of medial rectus muscle, the corrective extent was ( 4. 33 ± 0. 29 ) ? / mm. In over -correction cases with concomitant exotropia, 16 cases were performed advance of lateral rectus muscle, the corrective extent was (5. 37 ± 1. 56) ? / mm; 6 cases were performed recession of medial rectus muscle, the corrective extent was (6. 29 ± 3. 68) ? / mm; 5 cases were performed advance of lateral rectus muscle and recession of medial rectus muscle, the corrective extent was (5. 46±1. 78) ? / mm; 1 case were performed resection of lateral rectus muscle, the corrective extent was 5. 00? / mm. In under - correction cases with concomitant exotropia, 12 cases were performed resection of medial rectus muscle, the corrective extent was (4. 47 ± 0. 54) ? / mm; 16 cases were performed recession of lateral rectus muscle and resection of medial rectus muscle, the corrective extent was ( 5. 11 ± 0. 75 ) ? / mm; 2 cases were performed recession of lateral rectus muscle, the corrective extent was (2. 65±0. 42) ? / mm.
CONCLUSION: In reoperation of concomitant strabismus patients with over-correction, weakening or/and strengthening the horizontal muscle which were performed surgery before has a greater and more unstable surgical corrective extent. While In reoperation of concomitant strabismuspatients with under -correction, weakening or/ and strengthening the horizontal muscle which were not performed surgery has a normal corrective extent as usual.

AlM:To observe the changes of binocular vision in V-pattern exotropia children before and after surgical correction, and the effect of training in reconstructing the binocular vision after surgical corrections.METHODS: Sixty V-pattern exotropia children were enrolled in this study and were divided into three groups according to their age:group A (4~6 years old), group B (7~9 years old), and group C (10~12 years old), 20 cases for each group. Patients received routine refraction and ophthalmic examinations. Distance and near deviation were measured by prism-covering method and synoptophore. The simultaneous perception and fusion were examined with a synoptophore, and the stereacuity was measured with stereograms ( Titmus) . The children who didn’t reconstruct binocular vision function 1wk after surgery received binocular vision training. The data were recorded before and 1 , 2, 4, and 8wk after surgery. RESULTS: Binocular vision significantly improved among the children after surgery in group A and B ( P<0. 05 ) . Significantly divergence showed between group C and the other groups 1wk after surgery ( P < 0. 05 ). Binocular vision of the three groups all significantly improved 8wk after surgery, with no significant differences (P>0. 05). CONCLUSlON: V - pattern exotropia children can benefit from early surgical correction and training after surgery in reconstruct binocular vision.

Objective To investigate the relationship of antithrombin‐Ⅲ activity and thrombosis risk in liver cirrhosis with Child‐Pugh classification .Methods In our hospital from June to December 2014 ,60 liver cirrhosis patients were selected randomly included into this experiment group ,The 60 cases of control group were from medical examination of health in our hospital .The plasma AT‐Ⅲ activity and D‐D concentration in all these cases were detected and analyzed .Results The AT‐Ⅲ in cirrhosis patients were significantly lower than which in healthy persons(P<0 .05) .The lower level of AT‐Ⅲ is in these patients which were in seri‐ous condition(P<0 .05) ,the abnormal rate of D‐D concentration is also higher at the same time(P<0 .05) .Conclusion The detec‐tion of AT‐Ⅲ level in patients with liver cirrhosis is directly related to the severity of clinical and thrombosis risk .The AT‐Ⅲ de‐tection level can be used to judge the patient′s condition and develop appropriate treatment strategies .