OBJECTIVETo develop a simple, cheap and efficient restriction endonucleases fingerprinting-single strand conformation polymorphism(REF-SSCP) method applied to screen for mutations in long segments.

METHODSThe genomic DNA of Cx26 gene segment of the patients with deafness was amplified. The amplification products were screened with SSCP and REF-SSCP technique and DNA sequencing to evaluate and compare the effect on detection of mutations in long segments.

RESULTSNo different band was found in 724 bp segment in SSCP examination. Three kinds of different bands were discovered in REF-SSCP examination and the 79 G -->A mutation detected by DNA sequencing were accorded with the REF-SSCP bands entirely. The rate of detection was 100%.

CONCLUSIONThe present REF-SSCP method is applicable to screen mutations in long segment DNA of mass specimens.

Connexin 26 ; Connexins ; genetics ; DNA Fingerprinting ; methods ; DNA Restriction Enzymes ; Deafness ; genetics ; Genetic Testing ; methods ; Humans ; Mutation ; Polymorphism, Single-Stranded Conformational ; Sensitivity and Specificity

Objective To explore the neuroprotective mechanism of propofol by comparing the influence of propofol and isoflurane on inflammatory cytokines ( TNF-α、IL-1、ICAM-1 ) in patients with intracranial tumors. Methods One hundred and sixty-eight patients with intracranial neoplasm were randomly divided into two groups:the propofol ( Group P) and isoflurane (Group I),84 cases in each. Patients were given with propofol (3-6 μg·mL-1) by plasma target-controlled infusion or with continuously inhaled isoflurane ( 1%-2%) , respectively. The serum levels of TNF-α, IL-1 and ICAM-1 were detected before anesthesia and at 0,24,and 48 h after operation. Results The serum levels of TNF-α,IL-1 and ICAM-1 were significantly increased after operation as compared to baseline in both groups. The serum level of TNF-α was(69. 11±8. 95) and (76. 26±11.28) μg·mL-1,IL-1 was(21.57±3.19) and (29.58±4.38) ng·L-1,and ICAM-1 was (1.63±0.24)and (1.94±0.29) g·L-1 at 24 h post operation in Group P and Group I,respectively. These inflammatory cytokine levels were significantly higher in group I compared to group P at 24 and 48 h after operation (P<0. 05 or P<0. 01). Conclusion The target-controlled infusion of propofol brings about lower level of inflammatory reaction than isoflurane inhalation in patients with intracranial neoplasm,which may attribute to the mechanism of brain protection against injury.

Objective To investigate the effects of sevoflurane pretreatment on the inflammatory response in patients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB) and the mechanism of myocardial protection.Methods Twenty NYHA class Ⅱ or Ⅲ patients of both sexes,aged ＜ 60 yr,undergoing cardiac valve replacement with CPB,were randomly divided into 2 groups (n=10 each): sevoflurane group (group S) and control group (group C).The patients were premedicated with intramuscular morphine 0.1 mg/kg and scopolamine 0.3 mg.Anesthesia was induced with iv injection of midazolam 0.05-0.08 mg/kg,fentanyl 10-15 μg/kg and pipecuronium 0.08-0.10 mg/kg.The patients were tracheal intubated and mechanically ventilated.Anesthesia was maintained with intermittent iv boluses of midazolam0.03-0.06 mg/kg,fentanyl 5-10μg/kg and pipecuronium 0.04-0.08 mg/kg.Sevoflurane was inhaled before aortic clamping and the end-tidal concentration was rapidly adjusted to 1.0％ and maintained at this level for 30 min in group S.Blood samples were taken from the central vein before skin incision,immediately after aortic clamping,immediately after aortic unclamping and at 30 min after aortic unclamping,at 2,6,12 and 24 h (T1-8) after operation for determination of the concentrations of plasma tumor necrosis factor-α (TNF-α),interleukin-6 (IL) and interleukin-8 (IL-8),intercellular adhesion molecule-1 (ICAM-1),cardiac troponin I (cTnI) and activity of creatine kinase MB (CK-MB).The requirement for cardiovascular drugs was recorded after release of aortic cross clamp.Results Compared with group C,the plasma concentrations of TNF-α,IL-6 and IL-8 were significantly decreased at T3-8,the plasma concentrations of ICAM-1 and cTnl were significantly decreased at T4-8,the activity of plasma CK-MB was significantly decreased at T8,and the requirement for cardiovascular drugs was significantly reduced after release of aortic cross clamp in group S (P ＜0.05).Conclusion Sevoflurane pretreatment can inhibit the inflammatory response and provide myocardial protection to some extent in patients undergoing cardiac valve replacement with CPB.

Objective To investigate the effects of sevoflurane pretreatment on the myocardial injury in patients undergoing cardiac valve replacement with cardiopulmonary bypass (CPB).Methods Twenty NYHA class Ⅱ or Ⅲ patients,aged ＜ 60 yr,undergoing cardiac valve replacement with CPB,were randomly divided into 2 groups (n =10 each):sevoflurane group (group S) and control group (group C).The patients were premeditated with intramuscular morphine and scopolamine.Anesthesia was induced with iv injection of midazolam 0.05-0.08 mg/kg,fentanyl 10-15 μg/kg and pipecuronium 0.08-0.10 mg/kg.Anesthesia was maintained with intermittent iv boluses of midazolam,fentanyl and pipecuronium and in addition sevoflurane was inhaled before aortic clamping and the end-tidal concentration was rapidly increased to 1.0％ and maintained at the level for 5 min in group S.Blood samples were taken from the central vein before skin incision (T1),immediately after aortic clamping (T2 ),at 0 and 30 min after aortic unclamping (T3-4),and at 2,6,12 and 24 h after operation (T5-8) for determination of the concentration of serum cardiac troponin Ⅰ (cTnI) and activities of creatine kinase (CK) and creatine kinase isoenzyme-MB (CK-MB).Myocardial specimens were taken from right auricle before aortic clamping and at 10 min after aortic unclamping for electron microscopic examination.Results The concentration of serum cTnI and activities of CK and CK-MB were significantly increased at T4-8 in both groups ( P ＜ 0.05).The serum cTnI concentration at T4-8 and the activities of CK and CK-MB at T8 were significantly lower in group S than in group C ( P ＜0.05).Different degrees of mitochondrial swelling were observed after aortic unclamping in both groups,but the changes were milder in group S than in group C.Conclusion Sevoflurane pretreatment can attenuate the myocardial injury in patients undergoing cardiac valve replacement with CPB.

Cancer theranostics has attracted increasing attention in the area of nanooncology, where the therapeutic drugs and diag-nostic imaging are integrated into a multifunctional nanoplatform. A theranostic nanoparticle can deliver therapeutic drugs and imag-ing agents simultaneously. Iron oxide based magnetic nanoparticles (MNPs) are one of the most typical theranostic nanoparticles and have many excellent properties, such as biosafety, superparamagnetism, and tunable surface modifications and functionalizations. Moreover, they have drug loading capacity along with the distinctive properties of T2 weighted magnetic resonance imaging (T2W MRI), magnetic targeting, and magnetic hyperthermia. Presently, iron oxide based MNPs are being widely used in cancer theranostic research. This paper introduces the general structure of iron oxide based MNPs and reviews their applications in cancer dual/multiple modal imaging (T2W MRI combining T1W MRI, CT, optical imaging, PET/SPECT, and ultrasound) and therapy (chemotherapy, photody-namic therapy, photothermal therapy, and magnetic hyperthermia).

This study shows the case of a patient with peroneus brevis tendon rupture in ankle fracture. The patient com-plained of swelling, pain and activity limitation in her right ankle caused by sprain. She was diagnosed with ankle fracture, supina-tion-adduction (Lauge-Hansen typing) by medical history, physical examination and imaging. The peroneus brevis tendon was not-ed complete ruptured when suturing the incision after ORIF, which was then repaired by "8" shaped suture. Based on literature re-view, the injury mechanism of the case may be because of overload inversion force toward to lateral inferior from the fracture of fib-ula on the peroneus brevis tendon. Additional attention should be paid to patients who suffered from an supination-adduction ankle fracture for whether peroneus brevis tendon tears before and during surgery.

The objectives of this study were to prepare pitavastatin-loaded poly lactic-co-glycolic acid nanoparticles(PLGA), to characterize their pharmaceutical properties, to conduct in vitro drug-release from the nanoparticles, and to observe the effects on the proliferation of endothelial progenitor cells. Both pitavastatin-loaded PLGA and blank PLGA nanoparticles were prepared using emulsion-solvent diffusion method with PLGA being carrier materials. Morphology of the nanoparticles was observed by scanning electron microscopy(SEM), and particle size was analyzed by laser nanometer particle size analyzer. The drug loading and encapsulation efficiency were assayed using high-performance liquid phase. Impact of blank and pitavastatin-loaded nanoparticles on the viability of endothelial progenitor cells was investigated by CCK8 method. Pitavastatin-loaded PLGA nanoparticles exhibited the structure with spherical shape, smooth surface and average diameter of(230. 1±45)nm. The drug loading capacity and encapsulation efficiency were(10. 00±1. 83)% and(35. 54±5. 40)%, respectively. In vitro sustained-release of pitavastatin from the nanoparticles was found. The blank PLGA nanoparticles had no effect on the viability of the endothelial progenitor cells in different concentrations. Compared with pitavastatin group, pitavastatin-loaded nanoparticles(0. 01 μmol/L, 0. 1 μmol/L)had more effects on the proliferation of endothelial progenitor cells. In conclusion, emulsion-solvent diffusion method is applicable in preparation of pitavastatin-loaded PLGA nanoparticles with good shape and sustained-release of interest. Pitavastatin-loaded nanoparticles could significantly improve proliferation of the endothelial progenitor cells.

Objective: To evaluate the effect of zinc ions on human umbilical vein endothelial cells biological functions. Methods: The primary human umbilical vein endothelial cells were cultured with the ECM medium, and cells were divided into 8 groups: the control group(routine culture,n=3), 20 μmol/L zinc group(20 μmol/L zinc chloride solution was added into the cell medium, n=3), 40 μmol/L zinc group(40 μmol/L zinc chloride solution was added into the cell medium, n=3),80 μmol/L zinc group(80 μmol/L zinc chloride solution was added into the cell medium, n=3), 100 μmol/L zinc group(100 μmol/L zinc chloride solution was added into the cell medium, n=3), 200 μmol/L zinc group(200 μmol/L zinc chloride solution was added into the cell medium, n=3),300 μmol/L zinc group(300 μmol/L zinc chloride solution was added into the cell medium, n=3), 500 μmol/L zinc group(500 μmol/L zinc chloride solution was added into the cell medium, n=3). The cell proliferation curve was derived from real time cell analysis (RTCA). The viability value was obtained via CCK-8 reagent, and the migration distance was tested by scratch-wound assay while the adhesion function was detected by RTCA. Results: (1)After 18 hours, RTCA showed that the proliferation cell indexes were 4.5±0.6, 3.7±0.4, 3.6±0.3, 2.5±0.4, and 2.5±0.4 in the 20, 40, 80, 100, and 200 μmol/L zinc groups, as compared with 3.5±0.3 in the control group (all P<0.05). Proliferation cell indexes were 0 in both of the 300 μmol/L and 500 μmol/L zinc groups. (2)After 96 hours, the viability were 1.21±0.05, 1.10±0.03, 0.99±0.05, 0.62±0.02, 0.45±0.04, 0.11±0.01, and 0.12±0.06, respectively in the 20, 40, 80, 100, 200, 300, and 500 μmol/L zinc groups, as compared with 0.75±0.05 in the control group (all P<0.05). (3)After 12 hours, the migration distances were (0.56±0.11),(0.96±0.07),(0.49±0.02), and (0.29±0.01)mm in the 20, 40, 80, and 100 μmol/L zinc groups, as compared with (0.24±0.04)mm in the control group (all P<0.05). (4)After 18 hours, the adhesion cell index were 0.40±0.05, 0.31±0.01, 0.38±0.05, and 0.40±0.03 in the 20, 40, 80, and 100 μmol/L zinc groups, as compared with 0.24±0.04 in the control group (all P>0.05). Conclusions Zinc ions at lower concentration (≤80 μmol/L) can promote proliferation, viability and migration of human umbilical vein endothelial cells, but the adhesion function was not significantly affected by zinc ions. Zinc ions at higher concentration (≥200 μmol/L) can inhibit the cellular function of the human umbilical vein endothelial cells.

Objective:To compare the efficacy of modified single-incision and traditional double-incision for flexor hallucis longus tendon transfer in the treatment of Myerson type III chronic Achilles tendon ruptures in the middle-aged and elderly patients.Methods:A retrospective cohort study was conducted to analyze the clinical data of 106 patients with Myerson type III chronic Achilles tendon ruptures admitted to Ningbo No.6 Hospital from January 2015 to May 2021, including 54 males and 52 females; aged 48-82 years [(67.2±8.4)years]. Flexor hallucis longus tendon transfer was assigned to 60 patients using modified single-incision (modified single-incision group) and to 46 patients using traditional double-incision (traditional double-incision group). The operation time, intraoperative blood loss as well as American foot and ankle society (AOFAS) ankle-hindfoot score, foot and ankle outcome score (FAOS), visual analogue score (VAS) before operation, at postoperative 12 months and at the last follow-up were compared between the two groups. Time to recover walking and self-care ability, and incidence of complications were compared as well.Results:All patients were followed up for 16-52 months [(37.4±9.5)months]. The operation time and intraoperative blood loss were (51.6±7.4)minutes and (16.6±3.9)ml in modified single-incision group compared to (72.8±7.5)minutes and (32.9±5.2)ml in traditional double-incision group (all P<0.01). There was no significant difference in the AOFAS ankle-hindfoot score, FAOS and VAS between the two groups before operation (all P>0.05). At postoperative 12 months and at the last follow-up, AOFAS ankle-hindfoot score and VAS were not significantly different between the two groups (all P>0.05), but FAOS in modified single-incision group [(112.6±3.0)points, (114.4±3.1)points] was improved significantly compared with traditional double-incision group [(110.8±4.1)points, (112.7±4.3)points] ( P<0.05 or 0.01). At postoperative 12 months and at the last follow-up, the AOFAS ankle-hindfoot score, FAOS and VAS in both groups were improved or decreased significantly compared with those before operation (all P<0.05). All patients recovered their pre-injury daily activities. The time to recover walking and self-care ability were (9.6±2.0)weeks and (12.7±1.7)weeks in modified single-incision group compared to (10.8±1.8)weeks and (13.7±1.9)weeks in traditional double-incision group (all P<0.01). In modified single-incision group, superficial incision infection ( n=1) was found and cured after oral antibiotics. In traditional double-incision group, superficial incision infection ( n=2), deep incision infection ( n=1), deep venous thrombosis ( n=1) and medial plantar nerve injury ( n=1) were found and cured after symptomatic internal medical therapy; claw toe deformity ( n=2) was found and relieved after wearing customized insoles and functional exercise. The incidence of complications was 1.7% (1/60) in modified single-incision group compared to 15.2% (7/46) in traditional double-incision group ( P<0.01). Conclusion:In contrast with traditional double-incision surgery, modified single-incision for flexor longus tendon transfer in the treatment of Myerson type III chronic Achilles tendon ruptures in the middle-aged and elderly patients has advantages such as shorter operation time, less intraoperative blood loss, better functional recovery, faster postoperative recovery and less complications.