Objective:To establish a graded method to avoid mean fluorescence intensity(MFI)threshold of HLA Class I antibodies corresponding antigen,and the HLAMatchmaker program has been used to select the minimum mismatch value of donor-patient epitopes.Evaluate the application value of combining both methods in selecting HLA compatible platelets(PTL)for patients with immune platelet transfusion failure(IPTR)in improving platelet the corrected count increment(CCI).Methods:A total 7 807 PLT cross-matching compatible were performed by the solid-phase red cell adherence(SPRCA)method for 51 IPTR patients.The Luminex single antigen flow cytometry was used to detect HLA Class I antibodies in patients,and detected the MFI value for different specificity antigens of HLA Class I antibodies,was graded into strong positive group(MFI＞4 000,level 1),medium positive group(1 000＜MFI 4 000,2),weak positive group(500＜MFI≤1 000,3),and one negative control group(MFI≤500).The results of 7 807 SPRCA their negative/positive reaction wells were enrolled and statistically analyzed in different grades and the four groups,the statistical differences between the four groups were compared.Multiple applications for the select HLA Class I compatible donor events were made for patients in two cases,and HLAMatchmaker program was used to calculate the number of HLA Class I epitopes mismatches between the donors and patients.The donor with the minimum number of epitopes mismatches was selected,while avoiding the corresponding antigens of HLA Class I antibodies in levels 1 and 2,the provision of HLA compatible platelets for IPTR.After the transfusions,the CCI value of the platelet transfusion efficacy evaluation index was calculated,and the clinical evaluation of the transfusion effect was obtained through statistical analysis.Results:There were statistically significant differences in the positive results of SPRCA immunoassay among the strong positive group,medium positive group,and weak positive group of 51 IPTR patients with different specific of HLA-I class antibodies and corresponding antigens(all P＜0.001).The positive results showed a range from high to low,with strong positive group＞medium positive group＞weak positive group.There were a statistical difference among between the strongly positive or moderately positive groups and the negative control group(P＜0.001).There was no statistical difference between the weakly positive group and the negative control group(P＞0.05).The strong positive group was set as the corresponding specific HLA Class I site corresponding antigen grade 1 avoidance threshold,the medium positive group as the grade 2 avoidance thresholds,and the weak positive group as the grade 3 avoidance threshold.In the case of donor platelet shortage,it is not necessary to avoid the weak positive group.Avoiding the strategy of donor antigens and HLAMatchmaker program scores≤7 corresponding to HLA Class I antibodies of levels 1 and 2,with CCI values＞4.5 × 109/L within 24 hours,can obtain effective clinical platelet transfusion conclusions.Conclusion:When selecting HLA Class I compatible donors for IPTR patients,the grading avoids HLA Class I antibodies corresponding to donor antigens,and the donor selection strategy with the minimum scores of HLAMatchmaker program is comprehensively selected.The negative result confirmed by platelet cross-matching experiments has certain practical application value for improving platelet count in IPTR patients.

Objective To establish a method for rapid screening and quantifying the illegal addition of metolazone in tablet candy by ultra-high performance liquid chromatography-quadrupole-time of flight mass spectrometry(UPLC-Q-TOF/MS).Methods The component in samples was extracted with acetonitrile,then purified by Captiva EMR-Lipid purification column,detected by UPLC-Q-TOF/MS with Agilent RRHD Eclipse Plus C18 chromatographic column(100 mm×2.1 mm,1.8μm)under Targeted MS/MS mode.Results The linear regression analysis data for the calibration plots showed a good linear relationship over the concentration range of 50-1 000 ng·mL-1 for metolazone(r=0.999 0);the value of detection limit and quantification limit was found to be 1.0 μg.g1 and 2.5 μg·g-1.The average recovery rate was 98.15％(RSD=2.2％,n=18).Conclusion The method had the advantages of simple operation,qualitative and quantitative accuracy.It could be applied to daily inspection.

Objective: To explore the effect of P311 microspheres-loaded thermosensitive chitosan hydrogel on the wound healing of full-thickness skin defects in rats. Methods: The method of experimental study was adopted. The polyvinyl alcohol/sodium alginate microspheres (simple microspheres), P311 microspheres, and bovine serum albumin labeled with fluorescein isothiocyanate (FITC-BSA) microspheres were prepared by water-in-oil emulsification, and then their morphology was observed under a light microscope/inverted fluorescence microscope. Chitosan solution was prepared, chitosan solution and β-glycerol phosphate disodium hydrate were mixed to prepare simple thermosensitive hydrogels, and thermosensitive hydrogels loaded with simple microspheres or P311 microspheres were prepared by adding corresponding substances in simple thermosensitive hydrogels. The morphological changes of the prepared four liquids in the state of tilt was observed at 37 ℃. After being freeze-dried, the micromorphology of the prepared four liquids was observed under a scanning electron microscope. Eighteen 3-4-week-old male Sprague-Dawley rats were divided into normal group without any treatment, dressing group, chitosan group, hydrogel alone group, simple microspheres-loaded hydrogel group, and P311 microspheres-loaded hydrogel group, which were inflicted with one full-thickness skin defect wound on both sides of the back spine and were dealt correspondingly, with 3 rats in each group. Rats with full-thickness skin defects in the five groups were collected, the wound healing was observed on post injury day (PID) 0 (immediately), 5, 10, and 15, and the wound healing rates on PID 5, 10, and 15 were calculated. The wound and wound margin tissue of rats with full-thickness skin defects in the five groups on PID 15 and normal skin tissue in the same site of rats in normal group were collected, hematoxylin and eosin staining was conducted to observe the histological changes, immunohistochemical staining was performed to observe the expressions of CD31 and vascular endothelial growth factor (VEGF), and Western blotting was conducted to detect the protein expressions of CD31 and VEGF. The number of samples was all three. Data were statistically analyzed with one-way analysis of variance, analysis of variance for repeated measurement, and Bonferroni correction. Results: Simple microspheres were spherical, with loose and porous surface. The surfaces of P311 microspheres and FITC-BSA microspheres were smooth without pores, and the FITC-BSA microspheres emitted uniform green fluorescence. The diameters of the three microspheres were basically consistent, being 33.1 to 37.7 μm. Compared with chitosan solution and simple thermosensitive hydrogel, the structures of the two microspheres-loaded hydrogels were more stable in the state of tilt at 37 ℃. The two microspheres-loaded hydrogels had denser network structures than those of chitosan solution and simple thermosensitive hydrogel, and in the cross section of which microspheres with a diameter of about 30 μm could be seen. Within PID 15, the wounds of rats in the five groups were healed to different degrees, and the wound healing of rats in P311 microspheres-loaded hydrogel group was the best. On PID 5, 10, and 15, the wound healing rates of rats in dressing group and chitosan group were (26.6±2.4)%, (38.5±3.1)%, (50.9±1.5)%, (47.6±2.0)%, (58.5±3.6)%, and (66.7±4.1)%, respectively, which were significantly lower than (59.3±4.8)%, (87.6±3.2)%, (97.2±1.0)% in P311 microspheres-loaded hydrogel group (P<0.05 or P<0.01). The wound healing rates of rats in hydrogel alone group on PID 10 and 15, and in simple microspheres-loaded hydrogel group on PID 15 were (76.0±3.3)%, (84.5±3.6)%, and (88.0±2.6)%, respectively, which were significantly lower than those in P311 microspheres-loaded hydrogel group (P<0.05). The epidermis, hair follicles, and sebaceous glands could be seen in the normal skin of rats in normal group, without positive expressions of CD31 or VEGF. The wounds of rats in P311 microspheres-loaded hydrogel group on PID 15 were almost completely epithelialized, with more blood vessels, hair follicles, sebaceous glands, and positive expressions of CD31 and VEGF in the wounds than those of rats with full-thickness skin defects in the other four groups, and more protein expressions of CD31 and VEGF than those of rats in the other five groups. Conclusions: The P311 microspheres-loaded thermosensitive chitosan hydrogel can release the encapsulated drug slowly, prolong the drug action time, and promote wound healing in rats with full-thickness skin defects by promoting wound angiogenesis and re-epithelialization.
Rats ; Male ; Animals ; Hydrogels ; Vascular Endothelial Growth Factor A ; Chitosan/pharmacology* ; Serum Albumin, Bovine/pharmacology* ; Microspheres ; Polyvinyl Alcohol/pharmacology* ; Hematoxylin/pharmacology* ; Eosine Yellowish-(YS)/pharmacology* ; Rats, Sprague-Dawley ; Wound Healing ; Skin/injuries* ; Skin Abnormalities ; Soft Tissue Injuries ; Water/pharmacology* ; Alginates/pharmacology*

CD55 Antigens ; Complement Activation ; Humans ; Membrane Cofactor Protein ; Pemphigoid, Bullous ; Recombinant Fusion Proteins

OBJECTIVE: To investigate the molecular characteristics and intracellular growth ability of Legionella pneumophila (L. pneumophila) strains from 1989 to 2016 in Sichuan Province, China. METHODS: Seventy-nine isolates of L. pneumophila were collected from environmental and clinical sources, including cooling towers, hot springs, bath water, fountains, and patients, and identified with 16S rRNA gene analysis and serum agglutination assay. The isolates were then typed by Sequence-Based Typing (SBT), and Genotyping of forty-two LP1 strains were analyzed by means of multiple-locus VNTR analysis with 8 loci (MLVA-8). All strains were further analyzed for two virulence genes: Legionella vir homologue (lvh) and repeats in structural toxin (rtxA). The intracellular growth ability of 33 selected isolates was determined by examining their interaction with J774 cells. RESULTS: All isolates were identified to L. pneumophila including 11 serogroups, among which the main serogroup were LP1, accounting for 54.43%. Thirty-three different sequence types (STs) from five main clonal groups and five singletons were identified, along with 8 different MLVA patterns. Both the lvh and rtxA loci were found in all 79 strains. Thirty isolates showed high intracellular growth ability in J774 cells. CONCLUSION L. pneumophila is a potential threat to public health, and effective control and prevention strategies are urgently needed.
Bacterial Proteins ; genetics ; Bacterial Toxins ; genetics ; China ; Genotyping Techniques ; Humans ; Legionella pneumophila ; genetics ; growth & development ; isolation & purification ; RNA, Ribosomal, 16S ; genetics ; Water Microbiology

BACKGROUND: Multilevel thoracolumbar fractures are mainly treated with percutaneous pedicle screw and open pedicle screw system, but the treatment effect of different systems and the accuracy rate of screw placement are controversial, resulting in the lack of uniform standards for choosing the treatment method. OBJECTIVE: To evaluate the effect of percutaneous pedicle screw and open pedicle screw system in the treatment of multilevel thoracolumbar fractures and to evaluate the accuracy of the screw placement. METHODS: Totally 90 patients with multilevel thoracolumbar fractures were divided into open pedicle screw group (n=43 cases) and percutaneous pedicle screw group (n=47) according to different surgical methods. Open pedicle screw group was treated with open pedicle screw treatment, and percutaneous pedicle screw group was treated with percutaneous pedicle screw. Comprehensive effects were analyzed by comparing perioperative indicators (operation time, postoperative drainage volume, and incision length) imaging index (anterior vertebral height percentage, posterior vertebral height percentage, sagittal Cobb angle), postoperative complications, and pedicle screw accuracy. RESULTS AND CONCLUSION: (1) The amount of bleeding, postoperative drainage volume, and incision length were less (shorter) in the percutaneous pedicle screw group compared with the open pedicle screw group (P < 0.05). However, operation time and the number of undergoing fluoroscopy were longer (more) in the percutaneous pedicle screw group than in the open pedicle screw group (P < 0.05). (2) Anterior vertebral height percentage and posterior vertebral height percentage were higher in the percutaneous pedicle screw group than in the open pedicle screw group (P < 0.05). Sagittal Cobb angle was smaller in the percutaneous pedicle screw group than in the open pedicle screw group (P < 0.05). (3) At 2 months after surgery, the complication rate was significantly lower in the percutaneous pedicle screw group (4%) than in the open pedicle screw group (14%) (P < 0.05). (4) The accuracy rate of pedicle screw was significantly higher in the percutaneous pedicle screw group (92.1%; 279 screws) than in the open pedicle screw group (77.0%; 257 screws) (P < 0.05). (5) Results indicated that percutaneous pedicle screw fixation is characterized by less trauma and rapid recovery in the treatment of multilevel thoracolumbar fractures. It is helpful for the reduction of the injured vertebra, the maintenance of vertebral height; the safety and the accuracy of screw placement are high.

OBJECTIVETo compare the short term curative effect of posterior open door laminoplasty between continuous placement of shaping plate and intermittent placement in treating multilevel cervical spondylotic myelopathy.

METHODSFrom January 2012 to March 2015, 43 patients with multi segment cervical spondylotic cervical were treated with posterior open door laminoplasty, 21 patients with continuous placement of shaping plate(continuous group), 22 patients with intermittent placement of shaping plate(intermittent group). Operative time, intraoperative blood loss, JOA score, VAS score, postoperative spinal sagittal diameter and cervical curvature, postoperative cervical activity, complications, hospitalization expenses etc. were observed.

RESULTSThe patients of two groups were followed up with an average of (23.2±8.1) months and (23.3±8.0) months in continuous group and intermittent group, respectively. There was no significant difference in operative time, intraoperative blood loss, hospitalization time between two groups(>0.05). JOA and VAS scores of all patients at final follow up were obviously improved than preoperative(<0.05). Postoperative spinal sagittal diameter at 3 days and final follow up were obviously improved(<0.05), and there was no significant difference between postoperative at 3 days and final follow up(>0.05). Cervical activity of all patients at final follow up was decreased than preoperative(<0.05), but there was no significant difference between two groups(>0.05). There was no significant difference in postoperative complication and there was significant difference in hospitalization expenses between two groups.

CONCLUSIONSPosterior open door laminoplasty with continuous or intermittent placement of shaping plate have similar clinical effects in ameliorating nerve function for the treatment of multilevel cervical spondylotic myelopahty. However, the hospitalization expenses of intermittent group is obviously reduced, and the medical resources can be saved.

OBJECTIVETo investigate the clinical effects and prevent the complications of posterior and anterior decompression and internal fixation in the revision of cervical anterior internal fixation failure.

METHODSFrom 2008 January to 2011 December, 17 patients with cervical anterior internal fixation failure were treated with posterior and anterior decompression and internal fixation. There were 12 males and 5 females, aged from 26 to 68 years old with an average of 44.1 years. The lower screw loosening was found in 6 cases, the upper screw loosening in 5 cases, titanium mesh caving in 3 cases, the upper screw breakage in 2 cases, the lower screw breakage in 1 case. Informations of bone fusion were observed by X-ray, CT, MRI. Clinical effects were evaluated by modified JOA score.

RESULTSAll the revision operations were successfully completed. One case with poor blood coagulation function before operation resulted in postoperative hematoma and occurred neurological symptoms; after hematoma removal and fresh frozen plasma infusion later, neurological symptoms of the patient disappeared. All patients were followed up from 6 to 38 months with an average of (22.4±10.0) months. Postoperative at 2 weeks, 3 months, and final follow-up, JOA score had obviously improved and respectively was 13.1±1.6, 13.4±1.6, 14.2±1.5. All internal fixation locations were good after revision,and obtained bone fusion at 10 months after operation, with an average fusion time of 6 months.

CONCLUSIONThe combined posterior and anterior decompression and internal fixation in the revision of cervical anterior internal fixation failure is safe, can achieve thoroughly decompression, maintain the cervical curvature, reconstruct the three column stability, and it may be used for the patients of cervical anterior fixation failure.

Adult ; Aged ; Cervical Vertebrae ; surgery ; Decompression, Surgical ; methods ; Female ; Fracture Fixation, Internal ; methods ; Humans ; Male ; Middle Aged

BACKGROUNDIncomplete right bundle branch block (ICRBBB) is commonly associated with atrial septal defect (ASD), but lacks sufficient diagnostic test characteristics. An abnormal T wave is also often observed in ASD, with horizontal or inverted displacement of the proximal T wave limb in the right precordial leads, termed "defective T wave" (DTW).

METHODSWe examined the diagnostic test characteristics of combining ICRBBB with DTW as a new index to diagnose ASD. A total of 132 consecutive patients with ASD and 132 cases of age/gender-matched controls without ASD were enrolled.

RESULTSSensitivities of DTW, ICRBBB, and both were 87.1% - 87.9%. Specificities were 97.0%, 96.2%, and 100%, respectively. Positive predictive values were 1.3%, 1.1%, and 100.0% respectively, while negative predictive values were 99.9% for each.

CONCLUSIONCombining ICRBBB with DTW in electrocardiogram (ECG) as a new index significantly increased the specificity and positive predictive values while maintaining a high sensitivity in diagnosing ASD.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bundle-Branch Block ; diagnosis ; physiopathology ; Child ; Child, Preschool ; Electrocardiography ; Female ; Heart Septal Defects, Atrial ; diagnosis ; Humans ; Male ; Middle Aged ; Predictive Value of Tests ; Sensitivity and Specificity

BACKGROUNDThe genome of the severe acute respiratory syndrome-associated coronavirus (SARS-CoV) includes sequences encoding the putative protein X4 (ORF8, ORF7a), consisting of 122 amino acids. The deduced sequence contains a probable cleaved signal peptide sequence and a C-terminal transmembrane helix, indicating that protein X4 is likely to be a type I membrane protein. This study was conducted to demonstrate whether the protein X4 was expressed and its essential function in the process of SARS-CoV infection.

METHODSThe prokaryotic and eukaryotic protein X4-expressing plasmids were constructed. Recombinant soluble protein X4 was purified from E. coli using ion exchange chromatography, and the preparation was injected into chicken for rising specific polyclonal antibodies. The expression of protein X4 in SARS-CoV-infected Vero E6 cells and lung tissues from patients with SARS was performed using immunofluorescence assay and immunohistochemistry technique. The preliminary function of protein X4 was evaluated by treatment with and over-expression of protein X4 in cell lines. Western blot was employed to evaluate the expression of protein X4 in SARS-CoV particles.

RESULTSWe expressed and purified soluble recombinant protein X4 from E.coli, and generated specific antibodies against protein X4. Western blot proved that the protein X4 was not assembled in the SARS-CoV particles. Indirect immunofluorescence assays revealed that the expression of protein X4 was detected at 8 hours after infection in SARS-CoV-infected Vero E6 cells. It was also detected in the lung tissues from patients with SARS. Treatment with and overexpression of protein X4 inhibited the growth of Balb/c 3T3 cells as determined by cell counting and MTT assays.

CONCLUSIONThe results provide the evidence of protein X4 expression following SARS-CoV infection, and may facilitate further investigation of the immunopathological mechanism of SARS.

Amino Acid Sequence ; Animals ; BALB 3T3 Cells ; Cercopithecus aethiops ; Growth Inhibitors ; analysis ; physiology ; HeLa Cells ; Humans ; Immunohistochemistry ; Lung ; chemistry ; Mice ; Molecular Sequence Data ; SARS Virus ; chemistry ; Severe Acute Respiratory Syndrome ; metabolism ; Vero Cells ; Viral Structural Proteins ; analysis ; physiology