Objective To determine the effect of hypothermia on gene transcription and protein expression of calpain after traumatic brain injury (TBI). Methods Twenty-seven rats were randomly divided into three groups, ie, normal control group, normothermia TBI group and hypothermia TBI group. All rats with TBI were suffered from a lateral fluid percussion injury (FPI) at the right parietal lobe. Hy-pothermia intervention [rectal temperature for (32 ± 0.5) ℃] was performed for four hours immediately after TBI in hypothermia TBI group. Fluorescence PCR and Western blot were utilized to semi-quantify gene transcription and protein expression of ealpain and immunofluorescence used to observe protein dis-tribution of Calpain. Results Compared with normothermia TBi group and normal control group, hypo-thermia TBI group showed increased calpain gene transcription at 12 and 24 hours respectively after FPI (P <0.05). However, the increase of ealpain protein expression in hypothermia TBI group was inhibited more significantly by hypothermia at 6,12,24 and 72 hours after TBI, compared with normothermia TBI group (P < 0.05). Conclusion Neuroproteetion of hypothermia after TBI may somewhat be related to the decrease of calpain protein expression after its gene transcription.

Objective To investigate the changes of circulating endothelial progenitor cells (EPCs) in patients with acute cerebral infarction or chronic cerebral ischemia and discuss the related clinical significance.Methods Circulating EPCs were isolated using staining markers of CD34,CD133,and kinase insert domain receptor (KDR).Peripheral venous blood was collected from patients with acute cerebral infarction within 24 hours of onset (infarction group,n =30),with chronic cerebral ischemia (ischemia group,n =20),and without cerebral ischemia (control group,n =10) to quantify circulating level of EPCs using flow cytometry and measure parameters of systolic pressure,glycosylated hemoglobin (HbAlc),total cholesterol (TC),and triglyceride (TG),and low density lipoprotein-cholesterol (LDL-C),and high density lipoprotein-cholesterol (HDL-C).Results CD34-,CD34/CD133-,and CD34/KDR-positive cells counted (14.2 ± 8.1)‰,(7.1 ± 4.1)‰ and (5.0 ± 3.7)‰ in infarction group,(28.5 ± 9.9)‰,(15.2 ± 3.7)‰ and (6.8 ± 2.0)‰ in ischemia group,and (44.8 ± 9.5) ‰,(22.1 ± 6.6) ‰ and (16.7 ± 6.9) ‰ in control group.Taken together,circulating level of EPCs lowered substantially in infarction and ischemia groups compared to control group (P ＜ 0.05) and a far lower level was observed in infarction group (P ＜ 0.05).Circulating level of EPCs in infarction group was in a moderate negative correlation with systolic pressure,TC,TG,and LDL-C (P ＜ 0.05).Conclusions Decreased circulating level of EPCs may be a risk factor to the development of cerebral ischemia in acute cerebral infarction patients.Therefore,level of EPCs is vital for prediction,prevention and treatment of acute cerebral infarction.

Objective To explore the effects of mitogen-activated protein kinase (MAPK) pathway by estradiol induced vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) in endometrial cancer Ishikawa cells.Methods The experiments were divided into 4 groups:E2 group (Ishikawa cells treated with 1 p mol/L estradiol for 30 minutes); inhibitor group:including Ishikawa cells treated with 10 μmol/L Bibf1 120 (Bibf1 120 group),or treated with 2.5 μmol/L Ponatinib (Ponatinib group),or treated with 10 p mol/L U0126 (U0126 group) for 60 minutes; inhibitor + E2 group:including Ishikawa cells treated with 10 μmol/L Bibf1120 (Bibf1120 + E2 group),or treated with 2.5 μmol/L Ponatinib (Ponatinib + E2 group),or treated with 10 μmol/L U0126 (U0126+ E2 group) for 60 minutes following incubation with 1 μmol/L estradiol for 30 minutes; control group:only adding the culture medium without serum DMEM.(1) Western blot analysis was used to detect phosphorylation extracellular signal-regulated kinase 1/2(p-ERK 1/2) protein expression with stimulation in different concentrations of estradiol (0.01,0.1,1,10,100 μmol/L).(2) Quantitative fluorescent reverse transcription (qRT)-PCR and western blot analysis was used to test the level of mRNA and protein of VEGF,bFGF,MAPK kinase 1/2 (MEK1/2),extracellular signal-regulated kinase 1/2 (ERK1/2),p-ERK1/2 and phosphorylation MEK1/2 (p-MEK1/2).Flow cytometry were used to examine the cell cycle,and transwell chamber assay were used to detect the cell migration in different groups.Results The expression of the p-ERK1/2 protein at 0.01,0.1,1,10,100 μ mol/L were 0.16±0.03,0.10±0.03,0.41 ±0.04,0.19±0.03,0.19±0.03,there were significantly higher than that in control group(0.05±0.00,P＜0.05),and which was more obvious at the concentration of 1 μmol/L estradiol.The expression level of VEGF,bFGF mRNA and protein in E2 group were higher than those in the control group (P＜O.05).VEGF mRNA and protein in Bibf1120+E2 group were higher than those in E2 group.The expression of MEK1/2,ERK1/2 mRNA protein in E2 group were higher than those in control group (P＜0.05).The expression of MEK1/2,ERK1/2 mRNA or p-MEK1/2,p-ERK1/2 protein in Bibf1120 + E2 group,Ponatinib+E2 group or U0126+E2 group were lower than those in E2 group(all P＜0.05).Percentage of G1 phase [(53.6±3.2)％] and S phase[(29.2±4.2)％] in E2 group was significantly different with those in control group respectively(P＜0.05).Percentage of G1 phase[(66.8±2.6)％,(63.1±2.6)％ and (63.3±0.4)％] and S phase [(25.4±1.9)％,(25.0±3.8)％ and(23.8±0.5)％] in U0126+E2 group,Bibf1120+E2 group or Ponatinib +E2 group was also significantly different with those in control group (all P＜0.05); percentage of G1 phase and S phase in U0126+E2 group was significant difference with those in Bibf1120+E2 group or ponatinib+E2 group (P＜0.05).The number of cell colony in E2 group (110± 17) was more than those in control group (65±8) ;the number of cell colony in U0126+E2 group(28±4),Bibf1120+E2 group(38±5) or Ponatinib+E2group(42±6) were significant different with those in E2 group (P＜0.05),the number of cell colony in U0126+E2 group was significant difference with those in Bibf1 120+E2 group or Ponatinib+E2 group (all P＜0.05).The results shown that the abilities of proliferation and cell migration were significantly increased in cells after estradiol stimulation.Conclusion Estradiol inducing the production of VEGF and bFGF could activate MAPK pathway through ER-independent manner,further promote development.

Objective To investigate the efficacy of endovascular stenting for aortic arch artery stenosis after nasopharyngeal carcinoma radiotherapy. Methods The clinical data of 8 patients with symptomatic severe aortic arch artery stenosis after nasopharyngeal carcinoma radiotherapy were analyzed retrospectively. The patients were all received endovascular stenting,and their improvement of cerebral ischemic symptoms was observed. They were followed up by cervical color Doppler ultrasound.Results The whole brain vascular DSA confirmed that there were 24 severe arterial stenoses on the aortic arch arteries of extracranial segments in 8 patients,including 11 in internal carotid artery,2 in common carotid artery,10 in vertebral artery and 1 in subclavian artery. The patients were treated with vascular angioplasty and stenting respectively. All the patients were followed up for 1 year;there were no recurrence of cerebral ischemic symptoms.Cervical color Doppler ultrasound did not reveal any obvious restenosis. Conclusion Endovascular stent angioplasty for the treatment of aortic arch artery stenosis after nasopharyngeal carcinoma radiotherapy is relatively safe and feasible.

OBJECTIVETo study the histogenesis, pathologic features and differential diagnosis of prostatic primary signet ring cell carcinoma.

METHODS10 cases of the primary signet ring cell carcinoma were detected from 262 cases of prostate carcinomas diagnosed on needle biopsy were investigated by routine pathological, immunohistochemical and histochemical methods, and then compared with 10 cases of signet ring cell carcinomas of the stomach and colon. 3 cases were studied with electron microscopy.

RESULTS9 cases of prostate signet ring cell carcinoma were associated with concurrent high-grade conventional prostatic carcinoma, but at least 25% of the neoplasm consisted of signet ring cells. Only one case was pure signet ring cell carcinoma. Neoplastic signet ring cells may be divided into two types: the first type showed formation of intracytoplasmic lumina or vacuole, and the second type had intracytoplasmic accumulation of excess PSA and/or PAP. Both types of signet ring cells were negative for mucin staining (AB/PAS and mucicarmine). Therefore they differed from signet ring cell carcinomas of the stomach and colon.

CONCLUSIONSPrimary prostate signet ring cell carcinoma is a low-differentiated adenocarcinoma of a special histologic type, which arises from the epithelial cells of the prostate acinus. They can be distinguished from metastatic signet ring cell carcinoma of the stomach and colon and also from vacuolate degeneration of conventional carcinoma after endocrine or radiation therapy.

Carcinoma, Signet Ring Cell ; pathology ; Cell Differentiation ; Diagnosis, Differential ; Humans ; Immunohistochemistry ; Male ; Microscopy, Electron ; Middle Aged ; Prostatic Neoplasms ; pathology

Objective To evaluate the clinical value of autoantibody spectrum against ovarian cancer associated antigens combine CA125 in detecting and monitoring ovarian cancer. Methods Circulating IgG, IgM autoantibodies against ovarian cancer associated antigens which included TM4SF1, C1D,TIZ, OV-142,FXR1 and OV-189 were measured by indirect ELISA in serum from 126 patients with ovarian cancer (prior treatment), 42 patients with benign ovarian masses, 142 healthy women. Cut off value of IgG, IgM autoantibodies were determined by receive operating characteristic (ROC) curve. CA125 was measured in serum by immunoradiometric assay (IRMA). We evaluated the clinical value of combining multiple autoantibodies (autoantibody spectrum ), combining autoantibody spectrum with CA125 by binary logistic regresion. The positive ratio of autoantibody spectrum in serum (prior and post treatment ) of 24 synchronization patients with ovarian cancer was analyzed to evaluate the value in monitoring state of illness.Results Our data indicated that serum contains IgG, IgM autoantibodies against ovarian cancer associated antigens. The positive ratio of IgG autoantibodies in serum from ovarian cancer patients and cancer-free patients were 34. 1% - 47. 6% and 13.0% - 19. 0%, respectively ( P ＜ 0. 05 ). The positive ratio of IgM autoantibodies in serum from ovarian cancer patients and cancer-free patients were 39. 7% - 53.2% and 12. 0% -33.2%, respectively (P ＜0. 05). The positive ratio of IgG autoantibodies against FXR1 and IgM autoantibodies against TIZ,FXR1 and OV-189 in early stage ( Ⅰ - Ⅱ ) ovarian cancer(55.3% ,63.8%,61.7% and 66. 0% ) were significantly higher than those in advanced ( Ⅲ - Ⅳ )ovarian cancer( 34. 2%,39. 2% ,26. 6% ,45.6%; all P ＜ 0. 05 ). Combining five autoantibodies ( TM4SF1 IgG, TM4SF1 IgM, C1D IgG, FXR1 IgG and TIZ IgM ) showed significantly improved sensitivity (75.4%, P ＜ 0. 05 ), lower specificity (78. 3% ,P ＜ 0. 05 ) and similar accuracy (77. 1%, P ＞ 0. 05 ) in detecting ovarian cancer compared to those of CA125 (61.1% ,88.0% ,77. 1% ). But the autoantibody spectrum showed significantly improved sensitivity in classifying early stage (76. 6% ), compared to those of CA125 (51.1% ,P ＜ 0. 05 ).Combining autoantibody spectrum with CA125 showed significantly improved sensitivity ( 85.7% ), specificity (90. 8% )and accuracy (88.7%) in detecting ovarian cancer compared to those of autoantibody spectrum alone ( all P ＜ 0. 05 ), while CA125 ( 61.1%, P ＜ 0. 05; 88. 0%, P ＞ 0. 05; 77. 1%, P ＜ 0. 05 ). The positive ratio of combine the autoantibody spectrum with CA125 was significantly lower in 24 post-treatment serum (42%) compared to the pairing prior treatment serum ( 88%, P ＜ 0. 05 ). Conclusion Combining the autoantibody spectrum against ovarian cancer associated antigens with CA125 can improve sensitivity,specificity and accuracy in detecting early ovarian cancer and may be used to monitoring state of illness.

Objective To investigate the value of autoantibody of breast cancer susceptibility 1 associated RING domain (BARD1) splice variant (OV-142) in detection of ovarian cancer.Methods We cloned OV-142 gene into plasmid pET-30b(+).The recombinant protein of OV-142 was expressed in pET30b(+) system and purified. The autoantibody of OV-142 was detected by indirect enzyme-linked immunosorbent assay (ELISA).Results We successfully constructed the recombinant plasmid of OV-142.The recombinant protein was expressed in pET-30b(+) system and purified.The purification rate of the recombinant protein was up to 90%.The relative amount of autoantibody of OV-142 detected by indirect ELISA was analyzed by receiver operating characteristic curve (ROC) and the cutoff value was determined.Combination of the autoantibody IgG of OV-142 and CA125 was analyzed by logistic regression. The sensitivity,specificity and accuracy was 71.4%,89.1%,and 81.9%,respectively,which were higher than IgG (41.3%,84.2%,66.8% ) and CA125( 61.1%,88.0%,77.1% ) when used alone each.Conclusions OV-142 is a splice variant of BARD1.It may be a potential immunotherapy target of ovarian cancer.Detection of autoantibody of OV-142 is a potent complementary tool of CA125 in ovarian cancerdiagnosis.

Objective To investigate the correlation between the mean platelet volume (MPV) ,neutrophil-to -lymphocyte(NLR) and the severity as well as prognosis of sudden sensorineural hearing loss (SSHL) patients . Methods A retrospective cohort study involved 172 patients with SSHL from January 2012 to May 2015 .The distri-bution characteristics of routine blood (white blood cells ,neutrophil ,lymphocyte ,MPV ,NLR) in different audio-metric curves (hearing loss at low frequencies ,flat type ,high frequencies ,and total deafness) and prognosis of re-covery (complete ,partial ,slight ,and no recovery )were analyzed by SPSS 19 .0 analysis chi square test ,and the prognosis was estimated by using the receiver operating characteristic curve (ROC) .Results MPV and NLR levels in the severe and profound hearing loss group were significantly higher than that in different audiometric curves (P<0 .01 and P<0 .01) .MPV and NLR level in the partial recovery group and the no recovery group were signifi-cantly higher than that in the complete recovery group (P<0 .01 and P<0 .01) .The value of the MPV and NLR showed negative correlation with the prognosis (hearing recovery ) ,and the lymphocyte was positive with the prog-nosis .The sensitivity and specificity of MPV and NLR count 24 hours after admission predicting the prognosis of hearing recovery were 66 .2％ and 85 .5％ ,58 .4％ and 86 .7％ ,respectively .Conclusion The changes of MPV and NLR in SSHL patients are related to the severity of hearing loss ,and NLR count at 24 hours after admission may play an important role in prognosis of this disease .

Objective:To find out the optimal ratio of tip projection to nasal length for Chinese in order to help doctors to reduce the dissatisfactory rate of rhinoplasty.Methods:The authors retrospectively reviewed the records of patients with rhinoplasty from June 2015 to June 2018. The patients were classified into satisfactory, underprojected and overprojected groups. The average, maximum, minimum of tip projection (TP), nasal length (NL) and TP/NL, median of TP/NL of each group were measured. The results were used in later patients to assess their efficacy by comparing the dissatisfactory rates of patients before and after June 2018.Results:In satisfactory group, the mean of TP/NL was 0.63 with minimum 0.56, maximum 0.69, and median 0.64 in primary cases, while the mean of TP/NL was 0.63 with minimum 0.52, maximum 0.75, and median 0.64 in secondary cases. In underprojected group, the mean of TP/NL was 0.60 with median 0.58. In overprojected group, the mean of TP/NL was 0.64 with median 0.65. The dissatisfactory rates of patients before and after June 2018 were 8% and 2.2%, respectively ( P>0.05). Conclusions:The optimal ratio of tip projection to nasal length for Chinese is near 0.63-0.64. Combined with the desire of patient, it is possible for doctors to satisfy the patients by controlling tip projections between 2.5 cm to 2.8 cm.

Objective:To explore the relationship between expression levels of CLOCK mRNA and protein and the clinical characteristics of patients with nasopharyngeal carcinoma.Methods:The frozen tissue specimens from 33 patients with nasopharyngeal carcinoma in the Affiliated Tumor Hospital of Guizhou Medical University from 2018 to 2019 were collected. Seventeen cases of tissue specimens from patients with nasopharyngeal chronic inflammation in the Affiliated Hospital of Guizhou Medical University in 2019 were collected. From 2008 to 2014, 68 cases of formalin-fixed paraffin-embedding (FFPE) nasopharyngeal carcinoma tissue and 37 cases of FFPE nasopharyngeal chronic inflammation tissue were collected from the Affiliated Tumor Hospital of Guizhou Medical University. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) and western blot (WB) were used to detect the mRNA and protein expression levels of CLOCK. The nasopharyngeal carcinoma cells including CNE1, CNE2, 5-8F and the normal nasopharyngeal epithelial cell NP69 were cultured. qRT-PCR was used to detect the expression level of CLOCK mRNA in each cell line at the time points of ZT2, ZT6, ZT10, ZT14, ZT18 and ZT22. The cosine method was used to fit the rhythm of CLOCK gene in nasopharyngeal carcinoma. The protein expression of CLOCK protein was detected by using immunohistochemical method in 68 cases of nasopharyngeal carcinoma and 37 cases of nasopharyngeal chronic inflammation tissue. Survival was analyzed by Kaplan-Meier method and Log rank test, and the influencing factors was analyzed by Cox regression model.Results:The expression levels of CLOCK mRNA in CNE1, CNE2 and 5-8F cells (0.63±0.07, 0.91±0.02 and 0.33±0.04, respectively) were lower than that in NP69 cell (1.00±0.00, P<0.05). The expression levels of CLOCK protein in CNE1, CNE2 and 5-8F cells (0.79±0.06, 0.57±0.05 and 0.74±0.10, respectively) were lower than that of NP69 cells (1.00±0.00, P<0.05). The expressions of CLOCK mRNA in nasopharyngeal carcinoma cells including CEN1, CNE2, 5-8F and normal nasopharyngeal epithelial cell NP69 were different at different time points, with temporal fluctuations. The fluctuation periods of CLOCK mRNA in CNE1, CNE2, 5-8F, and NP69 cells were 16, 14, 22 and 24 hours, respectively. The peak and trough times were ZT10: 40 and ZT18: 40, ZT10 and ZT3, ZT14: 30 and ZT3: 30, ZT12: 39 and ZT0: 39, respectively. CLOCK mRNA and protein expression levels in nasopharyngeal carcinoma tissues (0.37±0.20 and 0.20±0.26, respectively) were lower than those in nasopharyngeal chronic inflammation tissues (1.00±0.00 and 0.51±0.41, respectively, P<0.05). The 1, 3, and 5-year survival rates of patients in the CLOCK protein high expression group (CLOCK protein expression level ≥ 0.178) were 96.2%, 92.1%, and 80.1%, respectively, which were higher than those in the low expression group (CLOCK protein expression level <0.178, 92.9% , 78.6% and 57.1%, respectively, P=0.009). The 1, 3, and 5-year progression-free survival (PFS) rates of patients in the CLOCK protein high expression group were 96.2%, 87.8%, and 87.7%, respectively, which were higher than those in the low expression group (92.7%, 82.2%, and 70.8%, respectively, P=0.105). Compared with the low-expression group (100.0%, 96.9%, and 90.0%, respectively), the 1, 3, and 5-year recurrence-free survival rates of patients in the CLOCK protein high expression group (100.0%, 95.7%, and 95.7%, respectively) were not statistically significant ( P=0.514). Compared with the low-expression group (92.7%, 82.2%, and 79.3%), the 1, 3, and 5-year survival rates without metastasis in the CLOCK protein high expression group (96.2%, 92.0%, and 92.0%, respectively) were not statistically significant ( P=0.136). CLOCK protein expression and T stage were independent prognostic factors of overall survival ( P<0.05). Conclusions:The expression of CLCOK is downregulated in the nasopharyngeal carcinoma cell and nasopharyngeal carcinoma tissues. Clock gene CLOCK is rhythmically expressed in the nasopharyngeal carcinoma cells and normal nasopharyngeal epithelial cells. Compared with normal nasopharyngeal epithelial cells, the fluctuation period of CLOCK in nasopharyngeal carcinoma cells is shortened. The overall survival of patients in the CLOCK protein high expression group is better than that of low expression group. The expression of CLOCK protein is an independent influencing factor for overall survival. CLOCK gene may be a potential tumor suppressor gene in the nasopharyngeal carcinoma.