Objective To study the effects of improved Wright-Giemsa staining. Methods 40 semen samples by both traditional and improved Wright-Giemsa staining.The morphological characteristics stained by two methods was ob-served. Results The defective rates of the middle and tail parts of sperm were 7.02%± 2.4%and 11.02%± 2.03%respec-tively in improved staining group;while those were 5.48%±2.8%and 8.05%±2.56%in traditional staining group. There was statistical significance between two staining methods (P < 0.05). There was no significant difference of normal sperm and sperm head between these two staining methods;however, improved staining method showed a much clearer nucleus and ac-rosome staining than that of traditional methods. Improved Wright-Giemsa staining is better for sperm, vaginal discharge and Trichomonas vaginalis. More importantly, axostyle and flagella were clearly stained with the improved method. Conclusion Improved Wright-Giemsa staining is simple and can be used in clinical diagnosis.

Objective:This paper aims to analyze the related effects of the low price medicine policy, and prob-lems in the implementation process. Methods:To retrieve an official website of the state food and medicine supervi-sion and administration, collect the package supplements of the low-price medicines and analyze their varieties according to their situations, and calculate he highest and lowest average daily use indicators for the medicine, etc. from January 2009 to August 2014. Results:(1) The low-price medicine list contains 533 kinds of standard medi-cines, and the coincidence rate with the essential medicines list is 51. 59%. Results also show that 96. 82% of the low-price medicines are incorporated into the national medical insurance directory. ( 2 ) The daily medicine cost difference of the maximum and minimum bidder price to the same medicine produced by different manufacturers ran-ges from 0. 01 to 30. 96 Yuan with 94. 76% of the western medicine dosage constituting the existing price rise space. (3) The daily medicine cost difference ranges from 0. 01 to 19. 35 Yuan with 92. 13% of the proprietary Chinese medicine varieties constituting the existing price rise space. Discussions:The low-price medicine varieties lack in the strict proof, the fact of low pricing the medicine has a two-way effect, and the connection between the low-price medi-cine administration policy and other policies is unclear.

The present study was undertaken to investigate the effect of human PMNs on the production of TNF-α by the human peripheral blood mononuclear cells (PBMCs) and to elucidate its tentative mechanism. Human PMNs and PBMCs were isolated from the venous blood of healthy donors by dextran sedimentation and density gradient centrifugation. In the presence of lipopolysaccharide (LPS), PMNs and PBMCs were cocultured at the ratio of 2:1 for 20 h and the concentration of TNF-α in the supernatant was measured by enzyme-linked immunosorbent assay. The binding rate of monocytes with the fluorescein isothiocyanate-labeled LPS (FITC-LPS) and the mean surface fluorescence intensity of monocytes were analyzed by flow cytometry. Results showed that PMNs were capable of inhibiting the TNF-α release from PBMCs (P＜0.05). PMNs suppressed the TNF-α release from PBMCs by 45% on average when PMNs and PBMCs cocultured at the ratio of 2:1. Paraformaldehyde-fixed PMNs still demonstrated the same inhibition (P＜0.05)，which proved that the inhibition was dependent on cell-to-cell contact and suggested that effector molecules responsible for this effect existed on the cell surface of PMNs. In the presence of PMNs, the binding rate of monocytes with the FITC-LPS and the mean surface fluorescence intensity of monocytes were not affected compared with PBMCs alone (P＞0.05). As incubation time was prolonged, the binding of FITC-LPS to monocytes increased (P＜0.05). Thus PMNs did not block the binding of LPS with monocytes. It was concluded that PMNs suppressed the TNF-α release from PBMCs via cell-to-cell interaction. In a cell-contact dependent manner, PMNs might interfere with the signal transduction pathway through which LPS activated PBMCs, thus attenuating the response of PBMCs to LPS and downregulating the TNF-α release.

The samples of muscular tissue from pork,beef and lamb which were closely related in the genetic relationship were analyzed by ultra performance liquid chromatography-tandem mass spectrometric (UPLC-MS) technique.The specific peptide biomarkers of pig meat species were found and confirmed.Proteins from three pure meat samples were extracted and digested using trypsin,the digested proteins were identified by UPLC-triple time-of-flight (TOF)-MS,and the total ion chromatogram (TIC) was searched and analyzed against the UniProt database.Three high abundant homologous proteins of three species and 8 potential peptide biomarkers of pork were found.A multiple reaction monitoring (MRM) QTRAP-MS method was established to confirm the specificity of potential peptide biomarkers.As a result,five peptide biomarkers of pig species meat were confirmed,three of which were not reported.

AIM: To investigate the effect of human polymorphonuclear leukocytes (PMNs) on the release of TNF-? by the human peripheral blood mononuclear cells (PBMCs) and to elucidate its mechanism. METHODS: Human PMNs and PBMCs were isolated from the venous blood of healthy donors by dextran sedimentation and density gradient centrifugation. After the cells were cocultured at the ratio of 2:1 in the presence of lipopolysaccharide (LPS), the concentration of TNF-? in the supernatant was measured by enzyme-linked immunosorbent assay. The binding rate of monocytes with the fluorescein isothiocyanate-labeled LPS (FITC-LPS) and the mean surface fluorescence intensity of monocytes were analyzed by flow cytometry. RESULTS: PMNs do not produce detectable TNF-? in the presence of LPS. PMNs were capable of inhibiting the TNF-? release from PBMCs ( P

Swine is an ideal model for diabetes studies. Insulin and insulin resistance are closely related with diabetes. To investigate the effect of SOCS-3 in insulin resistance, porcine primary adipocyte was treated with insulin (100 nmol/L) and dexamethasone (300 nmol/L) to induce insulin resistance. The simi-quantitative PCR results suggested that insulin increased GLUT4, PPARgamma and SOCS-3 gene expression in primary culture porcine adipocytes and no change of OB gene expression. Under insulin resistance conditions, SOCS-3 and OB gene expression were up-regulated, whereas GLUT4 and PPARgamma gene expression were down-regulated in primary porcine adipocytes. The overexpression of PPARgamma gene resulted in the increase of GLUT4 expression by insulin. Different expression levels of SOCS-3 determined the inhibitory effects of insulin signaling. Induction of insulin resistance by dexamethasone was not only due to inhibition of glucose transportation, but also repression of insulin signaling. SOCS-3 might be a potential gene to block the insulin resistance.
Adipocytes ; cytology ; metabolism ; Animals ; Cells, Cultured ; Dexamethasone ; pharmacology ; Glucose Transporter Type 4 ; biosynthesis ; genetics ; Insulin ; pharmacology ; Insulin Resistance ; Leptin ; biosynthesis ; genetics ; PPAR gamma ; biosynthesis ; genetics ; Suppressor of Cytokine Signaling 3 Protein ; Suppressor of Cytokine Signaling Proteins ; biosynthesis ; genetics ; Swine

Objective: To analyze the clinicopathologic characteristics of poorly-differentiated chordoma with INI1 loss in children and to discuss the differential diagnosis. Methods: The clinical, radiological, histopathological profiles and molecular pathologic characteristics of two pediatric poorly differentiated chordoma cases with INI1 loss were reviewed. Results: The patients were a girl and a boy. Both lesions involved the slope. Both patients were presented with progressive muscle weakness or neck pain. Radiological examination showed clivus bone destruction and compression of the brain stem and cervical spinal cord. Histologically, the tumor cells lacked typical organization and were associated with inflammatory cells infiltration. On high power field, the tumor cells were ovoid or fusiform with prominent atypia, vacuolated nuclei and prominent nucleoli. By immunohistochemistry, the tumor cells expressed cytokeratin, epithelial membrane antigen, brachyury and were negative for INI1. In both cases, INI1 gene deletion was detected by FISH. Conclusions Poorly-differentiated chordoma with INI1 loss mainly occurs in children. The morphology is different from classical chordoma.INI1 gene deletion is detectable by FISH. It can be distinguished from atypical teratoid/rhabdoid tumors and other neoplasms by the identification of nuclear brachyury expression. The loss of INI1 expression in poorly-differentiated chordoma might be associated with a poorly-differentiated morphology and an adverse prognosis.

Objective To investigate the clinical and radiological features of children with first attack of inflammatory central nervous system disorders and seropositivity to myelin-oligodendrocyte glycoprotein (MOG).Methods The clinical course,cerebrospinal fluid (CSF),MRI studies,MOG status and outcomes were retrospectively analyzed in children with first attack of inflammatory central nervous system disorders and seropositivity to MOG who were hospitalized in Children's Hospital of Fudan University from January 2016 to April 2017.Results Thirteen patients including 8 males and 5 females were included in this study,the ratio of male/female was 1.6∶ 1,and the median age was six years.Ten patients were diagnosed with acute disseminating encephalomyelitis,and three with clinically isolated syndromes.Seven patients had elevated CSF lymphocyte cells,and five patients had elevated CSF protein.All the patients' sera were tested for the anti-MOG IgG,which ranged from 1∶10 to 1∶100 with cell-based assay.MRI results showed that multiple anatomical areas were involved.Twelve patients had brain lesions,in which 10 patients had multiple lobes involved and four had tumefactive demyelinating lesions.The affected anatomical areas included white matters in 11 cases,thalamus/basal ganglias in nine,corpus callosums in three,brainstems in 10,spinal cord in five.The MRI features were characterized by hazy,bilateral lesions without clear boundaries.Clinical symptoms were fully restored in all the patients after treated with intravenous globulin and methyl prednisone.The average follow-up time was 8.9 months,and none of the patients had clinical recurrence.Conclusions MOG was associated with many kinds of inflammatory demyelinating diseases of central nervous system in children.Most of them were diagnosed with acute disseminating encephalomyelitis which has an acute or sub acute clinical course.The clinical manifestations of patients showed diversity.Multiple anatomical areas were involved,and treatment with intravenous globulin and methyl prednisone was effective in the acute phase.All of the patients had a favorable outcome.

Objective: To investigate the clinical features and genetic characteristics of patients with TBC1D24 gene mutations. Method: The clinical data of a patient with novel TBC1D24 compound heterozygous mutations from Children′s Hospital of Fudan University were collected, the related literature was searched from China National Knowledge Infrastructure, Wanfang Data Knowledge Service Platform, National Center from Biotechnology Information and Pubmed (up to April 2016) by using search terms "TBC1D24" "epilepsy" . The clinical features, electroencephalogram (EEG) and prognosis of the patients with TBC1D24 gene mutations were studied. Result: The patient was a boy with non-consanguineous healthy parents.He had an acute episode of focal continuous myoclonus lasting a few hours with consciousness preserved at the age of 3 months.Myoclonic jerks alternatively affected the eyelids, either the right or left limbs, sometimes triggered by fever or fatigue.The frequency was once 3-7 days.At the age of 6 months he was found to have myoclonus seizures with onset from a unilateral eyes lid and limb lasting 10 more minutes and subsequently affected four extremities or the trunk.They occurred once 3-4 months with perserved consciousness and lasted from several hours to up to ten more hours.They mostly disappeared during sleep.He had ataxia and mild mental retarding.Paroxysmal anomalies were not found on ictal traces.A novel compound heterozygous mutation of TBC1D24 gene, c. 730G>A, p.A244T and c. 1571G>C, p.R524P were found in the patient.Further study showed that c. 730G>A mutation was inherited from his father and c. 1571G>C from his mother. These two were not reported in public databases and predicted deleterious by Mutation Taster and polyphen-2.Literature relevant to TBC1D24 published all around the world was reviewed, no Chinese cases with TBC1D24 gene mutations had been reported. The total of 24 cases including the present case with TBC1D24 gene mutation were reported.Among them, 11 cases had compound heterozygous mutations and 13 cases had homozygous mutations.Ten mutations were identified, including 1 termination mutation, 1 frameshift mutation and 8 missense mutations. Conclusion TBC1D24 gene mutational analysis should be performed on patients with early-onset focal continuous myoclonus, if the etiology was unclear.

Objective:To investigate the efficacy of percutaneous balloon compression (PBC) in the treatment of trigeminal neuralgia (TN) and the risk factors of postoperative recurrence.Methods:A retrospective case-control study was conducted to collect the clinical data of 200 TN patients admitted to Hebei General Hospital from February 2018 to February 2019. According to different treatment methods, the patients were divided into operation group ( n=150) and conservative group ( n=50). The operation group received PBC treatment, and the conservative group received conservative treatment. The clinical efficacy of the two groups was recorded, including total effective rate, VAS, quality of life score (physical function, health status, social function, mental health). Patients treated with PBC were followed up for 4 years, and were divided into relapse group ( n=23) and non-recurrence group ( n=127) according to postoperative recurrence. The baseline data and laboratory indexes of the two groups were recorded, including fasting blood glucose, procalcitonin, C-reactive protein (CRP), interleukin-6 (IL-6), tumor necrosis factor-α(TNF-α). Measurement data with normal distribution were expressed as mean±standard deviation ( ± s), and t-test was used for comparison between groups. Chi-square test was used for comparison between count data groups. The factors related to postoperative recurrence of TN patients were first analyzed by univariate analysis, and then by multivariate Logistic regression analysis. A nomogram prediction model was constructed to predict postoperative recurrence, and its prediction efficiency was evaluated. The model was stratified by X-tile software to discuss its clinical application value. Results:The total effective rate, VAS of the operation group were 94.00%, 2.14±0.57, the conservative group were 78.00%, 3.87±1.16, and the difference between the two groups was statistically significant ( P< 0.05). The scores of physical function, health status, social function and mental health of quality of life in the operation group after treatment were 89.75±4.67, 90.36±5.68, 87.68±4.16, 88.79±5.09, the conservative group were 82.54±4.03, 84.67±4.28, 81.45±4.09, 80.69±4.89, and the difference between the two groups was statistically significant ( P< 0.05). Of 150 patients treated with PBC, 23 (15.33%) relapsed. Hypertension( OR=2.537, 95% CI: 1.069-6.019), diabetes( OR=5.179, 95% CI: 1.492-17.970), hyperlipidemia( OR=2.447, 95% CI: 1.227-4.883), CRP≥6.62 mg/L( OR=6.386, 95% CI: 1.738-9.854), IL-6≥19.55 ng/L( OR=8.028, 95% CI: 1.279-12.214), procalcitonin ≥1.13 ng/mL ( OR=7.615, 95% CI: 5.020-14.559), TNF-α≥4.56 ng/L( OR=6.226, 95% CI: 4.950-13.337) were independent risk factors for postoperative recurrence of PBC ( P<0.05). Based on the nomogram constructed by the above 7 risk factors, the decision curve showed that the net benefit rate was greater than 0 when the threshold probability was between 0.01 and 0.91. The X-tile software was used to divide the model into three levels of low (>21.6), medium (13.8-21.6) and high (<13.8) risk according to the Logistic risk score, and the postoperative recurrence probability of PBC was 10.87%, 20.74% and 64.04%, respectively. The recurrence rate of PBC in the high-risk group was significantly higher than that in the medium-risk group and the low-risk group ( χ2=5.136, P=0.015). Conclusions:Percutaneous balloon compression of trigeminal nerve in the treatment of patients with trigeminal neuralgia has the advantages of high total effective rate, fewer postoperative complications, good clinical efficacy, quick effect and so on. The construction of percutaneous balloon compression of trigeminal nerve in patients with trigeminal neuralgia postoperative recurrence risk prediction model to provide reference for clinical improvement of patient rehabilitation.