Objective To investigate the application of leukoreduction filter in removal of leukocytes in platelet concentrate . Methods Platelet concentrate was prepared by using platelet‐rich plasma method .35 bags of the same type of prepared platelet concentrate were filtered by using leukoreduction filter .The changes of conventional indicators of platelet before and after filter were measured and recorded .The activating platelets indicators PAC‐1 and CD62p were detected by using flow cytometry .The platelet hypotonic shock was measured by biochemical analyzer .The platelet aggregation was measured by using platelet aggrega‐tion instrument .Results After platelet concentrate was filtered by using leukoreduction filter ,leukocyte removal rate was(98 .28 ± 0 .97)% ,platelet recovery rate was(86 .37 ± 2 .84)% .After filtration ,white blood cell count ,platelets ,red blood cells were reduced than before filtration(P<0 .05) .The differences of capacity of platelets ,mean platelet volume and pH before and after filtration were not statistically significant(P> 0 .05) .Before and after filtration ,the expression of platelet activation markers PAC‐1 and CD62p ,platelet aggregation and platelet hypotonic shock were not statistically different(P>0 .05) .Conclusion Platelet leukore‐duction filter could effectively filter leukocytes in platelet concentrate .It would not change the conventional indicators ,and not affect platelet activation ,aggregation and anti‐hypotonic shock capacity significantly .

Influenza A/H1N1 virus-encoded nonstructural, or NS1, protein inhibits the 3'-end processing of cellular pre-mRNAs by binding the cellular protein: the 30-kDa subunit of CPSF (cleavage and polyadenylation specificity factor, CPSF30). CPSF30 binding site of the NS1 protein is a potential target for the development of drugs against influenza A/H1N1 virus. A yeast two-hybrid screening system was constructed and used for screening Chinese medicines that inhibit the interaction of the A/H1N1 flu NS1 protein and human CPSF30 protein. The NS1 gene of A/H1N1 virus was amplified by consecutive polymerase chain reaction (PCR), and the human CPSF30 gene of HeLa cell cloned by reverse transcriptase-polymerase chain reaction (RT-PCR). Then the two gene fragments confirmed by sequencing were subcloned into the yeast expression vectors pGBKT7 and pGADT7, respectively. The two constructs, bait vector pGBKNS1 and prey vector pGADCPSF, were co-transformed into yeast AH109. The eight individual yeast colonies were picked and subjected to verification by PCR/gel electrophoresis. The inhibition of the NS1-CPSF30 interaction was allowed the identification of selective inhibitors. The four of more than thirty identified Chinese medicines, including 'Shuanghuanglian oral liquid', showed the strong inhibition of the NS1-CPSF30 interaction.

AIM: To investigate the ocular complication after radiation therapy for nasopharyngeal carcinoma(NPC).METHODS: The authors performed a previous study on keratopathy in 213 NPC patients who received first stage radiation and had at least 10 months of follow-up. These patients were categorized into three groups depending on NPC clinic stages. Rates and proportions of keratopathy occurring in these groups were compared and analyzed with Chi-square Test and Spearman rank correlation coefficient.RESULTS: Radiation keratopathy developed in 19 patients, about 8.9% (19/213). The latency value was 3 to 30days. The effect of NPC clinic stages and radiation did on the development of keratopathy was not statistically significant (P＞0.05).CONCLUSION: The NPC clinic stages and radiation doses plays few effects on the development of keratopathy. It may play a key role that corneal nerves damage induced ocular surface diseases. It can not be excluded that individuals have different sensitivities to radiation.

Objective To explore the diagnoses of spontaneous intracranial hypotension,and discuss the therapeutic efficacy of epidural blood patch therapy in spontaneous intracranial hypotension patients.Methods The clinical data of 12 patients with spontaneous intracranial hypotension,admitted to our hospital from January 2013 to December 2018,were retrospectively analyzed.The lumbar puncture results,MR imaging features of the skull and spine,and CT myelography (CTM) features of these patients were analyzed.The treatment efficacies of epidural blood patch,which included blind epidural blood patch and targeted epidural blood patch,were compared.Results The cerebrospinal fluid pressure of 12 patients was ≤ 60 mmH2O.Ten patients (83.3％) showed subdural fluid collections,enhancement of the pachymeninges,engorgement of venous structures,pituitary hyperemia,and sagging of the brain on brain MR imaging,and one of the patient showed pituitary hemorrhage.Seven patients (63.3％) showed spinal dural epithelial fluid accumulation and venous plexus expansion on spine MR imaging,and one of the patient showed dorsolateral dural discontinuous thickening of T6 and forward movement of the spinal cord caused by massive dorsal epidural effusion.Twelve patients in this group underwent CTM,and were found cerebrospinal fluid leakage.Twelve patients applied 14-times epidural blood tests;4 responded well to one-time targeted epidural blood patch therapy,with success rate of 100％;8 patients used blind epidural blood patch therapy,and 6 patients responded well to one-time therapy,with success rate of 75％,one patient improved with blind epidural blood stick twice,and one patient was ineffective twice.Conclusions Head MR imaging combined with spinal MR imaging is a non-invasive method to diagnose spontaneous intracranial hypotension.Myelogram can determine whether there is a leakage of spinal cerebrospinal fluid and accurately locate the leakage site.Epidural blood patch therapy is an effective method for treatment of patients with spontaneous intracranial hypotension.With the precise location of leak points by myelography,targeted epidural blood patch is more effective.

OBJECTIVETo study the expression of hTERT mRNA and Mad1 protein in lung cancer of Gejiu and Xuanwei and normal lung tissue and to investigate their correlations with lung cancer.

METHODSMad1 protein was detected by immunohistochemistry S-P method, and hTERT message RNA (mRNA) was detected by in situ hybridization (ISH) in 40 specimens of lung cancer of Gejiu Tin miners and 20 specimens of lung cancer of Xuanwei peasants and 20 specimens of normal lung tissue. The positive signals were quantitatively analyzed by HPIAS-100.

RESULTSThe positive unit (PU) of Mad1 protein was 16.77 +/- 6.01 in Gejiu Tin Miners lung cancer group, and 19.36 +/- 4.54 in Xuanwei peasant lung cancer group, compared with the normal lung tissue (46.05 +/- 7.26). The difference was highly significant (P < 0.01); The PU of hTERT mRNA was 72.10 +/- 13.07 in Gejiu Tin miners lung cancer group, and 74.20 +/- 15.17 in Xuanwei peasant lung cancer group, which was higher than that in normal tissue group (10.70 +/- 2.21). The difference was significant (P < 0.01). The expression of Mad1 protein was negatively correlated with the expression hTERT mRNA (P < 0.05, r = 0.9881, r = -0.999).

CONCLUSIONReduced hTERT mRNA expression may play an important role in the occurrence of lung cancer. The expression of hTERT mRNA and deletion of Mad1 protein are closely related to pathogenesis of lung cancer.

Cell Cycle Proteins ; metabolism ; China ; Female ; Humans ; Lung ; metabolism ; Lung Neoplasms ; metabolism ; Male ; Mining ; Nuclear Proteins ; metabolism ; Telomerase ; metabolism

OBJECTIVETo compare the clinical effects and radiographic outcomes of mini-open trans-spatium intermuscular and percutaneous short-segment pedicle fixation in treating thoracolumbar mono-segmental vertebral fractures without neurological deficits.

METHODSFrom August 2009 and August 2012, 95 patients with thoracolumbar mono-segmental vertebral fractures without neurological deficits were treated with short-segment pedicle fixation through mini-open trans-spatium intermuscular or percutaneous approach. There were 65 males and 30 females, aged from 16 to 60 years old with an average of 42 years. The mini-open trans-spatium intermuscular approach was used in 58 cases (group A) and the percutaneous approach was used in 37 cases (group B). Total incision length, operative time, intraoperative bleeding, fluoroscopy, hospitalization cost were compared between two groups. Visual analog scale (VAS) and radiographic outcomes were compared between two groups.

RESULTSAll patients were followed up from 12 to 36 months with an average of 19.6 months. No complications such as incision infection, internal fixation loosening and breakage were found. In group A, fluoroscopy time was short and hospitalization cost was lower than that of group B (P<0.05). But the total incision length in group B was smaller than that of group A (P<0.05). There was no significant differences in operative time, intraoperative bleeding, postoperative VAS and radiographic outcomes between two groups (P>0.05). Postoperative VAS and radiographic outcomes were improved than that of preoperative (P<0.05).

CONCLUSIONThe mini-open trans-spatium intermuscular and percutaneous short-segment pedicle fixation have similar clinical effects and radiographic outcomes in treating thoracolumbar mono-segmental vertebral fractures without neurological deficits. However, in this study, the mini-open trans-spatium intermuscular approach has a short learning curve and more advantages in hospitalization cost and intraoperative radiation exposure times, and is recommendable.

Adolescent ; Adult ; Female ; Fluoroscopy ; Fracture Fixation, Internal ; methods ; Humans ; Lumbar Vertebrae ; injuries ; surgery ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; Spinal Fractures ; surgery ; Thoracic Vertebrae ; injuries ; surgery ; Visual Analog Scale

OBJECTIVETo study the effect of adriamycin combined with hyperthermia on tumor formation and growth of human B lymphoma cell line (Raji) in nude mice.

METHODSTwenty-four BALB/C nude mice were divided into control group (37 degrees celsius;), chemotherapy group (37 degrees celsius;+ADM), hyperthermia group (42 degrees celsius;) and thermochemotherapy group (42 degrees celsius;+ADM), and in each mouse, 5 x 10(6) Raji cells were injected subcutaneously. The time and rate of tumor formation were observed, the tumor diameter was measured every 3 days and the tumor volume calculated to obtain the growth curves of the tumors. Three weeks after tumor formation, all the mice were executed to observe the histopathological changes of the tumor with HE staining.

RESULTSThe time of tumor formation in the control, chemotherapy, hyperthermia and thermochemotherapy groups were 11.2-/+1.7, 20.2-/+2.3, 15.3-/+1.6 and 23.8-/+1.7 days, respectively. Three weeks after tumor formation, the average weight of the tumors were 3.33-/+0.57, 2.26-/+0.28, 2.76-/+0. 26 and 1.73-/+0. 33 g, respectively, and the tumor growth inhibition rate was 48.0% in the thermochemotherapy group.

CONCLUSIONThermochemotherapy can be effective in inhibiting tumor growth and provides an alternative of adjuvant therapy for malignant B cell lymphoma.

Animals ; Antigens, CD20 ; metabolism ; Cell Line, Tumor ; Cell Transformation, Neoplastic ; Combined Modality Therapy ; Doxorubicin ; pharmacology ; therapeutic use ; Female ; Gene Expression Regulation, Neoplastic ; drug effects ; Humans ; Hyperthermia, Induced ; Leukocyte Common Antigens ; metabolism ; Lymphoma, B-Cell ; drug therapy ; genetics ; pathology ; therapy ; Male ; Mice ; Mice, Nude ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Xenograft Model Antitumor Assays

OBJECTIVETo explore the influence of concealed penis on the morphology of the corpus cavernosum in rats.

METHODSRat models of concealed penis were established by intra-pocket-suture of the root of the penis. Fifty rats were equally assigned to Groups A (2-month) and B (4-month), each further divided into a buried (n = 15) and a normal subgroup (n = 10). Changes in the morphology of the penile cavernous tissue were observed under the light microscope and transmission electron microscope.

RESULTSCompared with Group A, Group B showed significant ultra-structural pathological changes in the corpus cavernosum, including abnormal arrangement of endothelial and smooth muscle cells, massive hyperplasia of interstitial tissues, narrowed cavernous sinus, atrophic smooth muscle cells, degenerated mitochondria, dilated endoplasmic reticula, decreased dense bodies and contractile fibers, and cytoplasmic vacuolization. No significant differences were found in the appearance and weight of the corpus cavernosum between the buried and normal groups (P > 0.05).

CONCLUSIONConcealed penis does not significantly affect the appearance and weight of the corpus cavernosum, but causes ultra-structural pathological changes in it with the lengthening of time.

Animals ; Male ; Penis ; pathology ; ultrastructure ; Phimosis ; pathology ; Rats ; Rats, Sprague-Dawley

Measurement of platelet-associated imunoglobulin (PAIg) has frequently been applied for the diagnosis of idiopathic thrombocytopenic purpura (ITP) and other immune thrombocytopenias. In the present study, a flow cytometry (FCM) analysis has been used to detect and characterize PAIg in 47 patients with ITP and Evans' syndrome, 13 patients with non-immune thrombocytopenia, 10 patients with autoimmune hemolytic anemia (AIHA) whose platelet counts were in normal range, and 31 healthy volunteers. With FCM measurement, mean fluorescence intensity (MFI) of platelets from patients with ITP and Evans' syndrome (2.26 +/- 2.29) was significantly higher than those from non-immune thrombocytopenia (0.33 +/- 0.39), AIHA (0.17 +/- 0.07) and control subjects (0.25 +/- 0.15) (P < 0.01). Meanwhile, the percentage of positive platelets of patients with ITP and Evans' syndrome [(44.1 +/- 29.0)%] was also higher than those of non-immune thrombocytopenia [(17.5 +/- 9.4)%], AIHA [(10.7 +/- 7.5)%] and control subjects [(16.6 +/- 8.4)%] (P < 0.01). In addition, some peak shape abnormality appeared (double peaks and peak tail) in the histogram of fluorescence intensity (log) of 11 patients (23.4%) with ITP and Evans' syndrome either alone or accompanied with quantitative alteration of MFI and/or positive platelet percentage. In seven cases, the peak shape abnormality was the unique characteristic that could be detected and have never been seen in normal platelets. This phenotypic alteration perhaps reflects the existence of different platelet populations and could be of diagnostic value. Totally, the positive result of FCM measurement in patients with ITP and Evans' syndrome was 87.2%, slightly higher than 83.0% positive rate with ELISA method, without statistical difference. The correspondent rate of the results of these two analytical settings was 85.1%. This study shows that FCM assay is a rapid and sensitive method for the measurement of PAIg and seems to be suitable as a novel routine diagnostic technique of immune thrombocytopenia.

Objective To establish the chronic low flow myocardial hibernation animal model in pigs, and to assess the diagnostic value for myocardial hibernation by using various imaging methods. Methods A total of 13 miniswine (30-40 kg) were used. All animals underwent general anesthesia and orotracheal intubation while the animals were mechanically ventilated. Under sterile conditions, left ventriculography and coronary angiography were performed by introduction of catheter into the right femoral artery. Further, a left anterolateral thoracotomy was performed in the third intercostal space. The proximal LCX was dissected free to allow placement of an ameroid constrictor. More than 1 month later, left ventriculography and coronary angiography were performed again, followed by cine MRI at rest and during stress with low dose of dobutamine (5 ?g?kg -1 ?min -1 ), respectively. Traditional and/or breath hold cine MRI were used to evaluate regional left ventricular wall motion, corresponding to basal, midventricular and apical short axis tomograms. Regional wall motion score index (WMSI) was calculated. At the same time 99m Tc MIBI myocardial SPECT was performed at rest and during nitroglycerin administration, respectively. All animals were finally sacrificed for pathological examination. Triphenyl tetrazolium chloride (TTC) staining was used to assess the myocardial infarction. Electron microscopy was used to identify myocardial cellular changes characteristic of hibernating myocardium. Results Three pigs died during surgery or within two weeks after surgery. One pig died of anesthesia during SPECT examination, 1 pig suffered from aneurysm, and another one pig showed negative findings. The other 7 pigs were found with hypokinetic ( n =4) or akinetic ( n =3) myocardial regions related to stenosed LCX (70%-99%). Resting cine MRI demonstrated decreased regional motion of the lateral and posteroinferior walls (ischemic regions) of the left ventricle ( n =7), compared with the nonischemic anteroseptal regions; but the low dose dobutamine (5 ?g?kg -1 ?min -1 ) could recover those hypokinetic or akinetic myocardial regions, characteristic of hibernating myocardium. Resting 99m Tc MIBI myocardial SPECT ( n =6) showed a fixed perfusion defect on the corresponding ischemic areas, which became reversible on the nitrate augmented myocardial perfusion imaging. It also indicated myocardial viability presented at the ischemic areas. TTC staining revealed patchy infarction of the area at risk localized to the endocardial surface ( n =3), and no myocardial infarction ( n =4). Electron microscopy of sections from the hibernating regions revealed loss of contractile materials, increased numbers of small mitochondria, and glycogen accumulation within viable cardiomyocytes, which had been described as hallmarks of hibernating myocardium. Conclusion Chronic low flow myocardial hibernation can be reproduced in an animal model during progressive coronary stenosis caused by ameroid constrictor.