Hepatocellular carcinoma(HCC)is difficult to detect in its early stages and current treatment methods are associated with significant side effects and a high risk of developing drug resistance.This study aims to investigate the effect of phycocyanin(PC)on the apoptosis of human HCC HepG2 cells and its potential mechanism.HepG2 cells were treated with PC at concentrations of 0.1,0.25,0.5,1,2.5,5,and 10 μg/mL for 12 h,and with 10 μg/mL PC and 2.5 μmol/L Wip1 inhibitor(Wip1i)alone or in combination for 12 and 24 h,respectively.Cell proliferation levels were assessed using the CCK-8 cell proliferation-toxicity assay kit.Apoptosis levels were measured by Annexin V-FITC/Propidium Iodide double staining combined with flow cytometry.TMT(Tandem Mass Tag)proteomics quantitative technol-ogy was applied to analyze differential protein expression.Western blotting was used to detect the expres-sion levels of Wip1,p53,and phosphorylated-p53(Ser15)proteins.The CCK-8 assay revealed that PC effectively inhibited HepG2 cell proliferation in a concentration-dependent manner,with a half-maximal inhibitory concentration(IC50)of 19.37 μg/mL.Flow cytometry results showed that PC significantly in-duced apoptosis,with an apoptosis rate of 30.40％.Quantitative proteomics analysis indicated that PC induced activation of the p53 pathway.The CCK-8 assay showed that Wip1i enhanced the cytotoxic effect of PC on HepG2 cells.Western blotting confirmed that PC inhibited Wip1 expression,induced p53 pro-tein phosphorylation,and promoted the expression of total p53 protein.Additionally,Wip1i further en-hanced PC-mediated activation of the p53 pathway,increasing the expression of p53 and pP53(S15).In conclusion,PC may induce apoptosis by inhibiting the activity of the p53 negative regulator Wip1,thereby promoting apoptosis through the Wip1/p53 pathway.

Hepatocellular carcinoma(HCC)is difficult to detect in its early stages and current treatment methods are associated with significant side effects and a high risk of developing drug resistance.This study aims to investigate the effect of phycocyanin(PC)on the apoptosis of human HCC HepG2 cells and its potential mechanism.HepG2 cells were treated with PC at concentrations of 0.1,0.25,0.5,1,2.5,5,and 10 μg/mL for 12 h,and with 10 μg/mL PC and 2.5 μmol/L Wip1 inhibitor(Wip1i)alone or in combination for 12 and 24 h,respectively.Cell proliferation levels were assessed using the CCK-8 cell proliferation-toxicity assay kit.Apoptosis levels were measured by Annexin V-FITC/Propidium Iodide double staining combined with flow cytometry.TMT(Tandem Mass Tag)proteomics quantitative technol-ogy was applied to analyze differential protein expression.Western blotting was used to detect the expres-sion levels of Wip1,p53,and phosphorylated-p53(Ser15)proteins.The CCK-8 assay revealed that PC effectively inhibited HepG2 cell proliferation in a concentration-dependent manner,with a half-maximal inhibitory concentration(IC50)of 19.37 μg/mL.Flow cytometry results showed that PC significantly in-duced apoptosis,with an apoptosis rate of 30.40％.Quantitative proteomics analysis indicated that PC induced activation of the p53 pathway.The CCK-8 assay showed that Wip1i enhanced the cytotoxic effect of PC on HepG2 cells.Western blotting confirmed that PC inhibited Wip1 expression,induced p53 pro-tein phosphorylation,and promoted the expression of total p53 protein.Additionally,Wip1i further en-hanced PC-mediated activation of the p53 pathway,increasing the expression of p53 and pP53(S15).In conclusion,PC may induce apoptosis by inhibiting the activity of the p53 negative regulator Wip1,thereby promoting apoptosis through the Wip1/p53 pathway.

Objective To explore astrocyte activation's impact on cognitive deficits post-traumatic brain injury(TBI).Methods SPF male rats were divided into control,sham-operation and model groups with 10 rats per group.Control group did not receive the surgical intervention;sham-operation group underwent craniotomy without damaging the dura mater;model group was struck with brain injury instrument after craniotomy.The rats with knockout overexpressed glial fibrillary acidic protein(GF AP)gene were set as the knockout group and overexpression group,with 10 rats per group.The knockout and overexpression groups were treated as the model group.Neurological function was evaluated by modified neurological severity score(mNSS).Escape latency was assessed with the Morris water maze test.The expression levels of GFAP in astrocytes were measured quantitative real-time polymerase chain reaction.The positive expression of astrocytes was determined by immunohistochemistry.Results The mNSS of control,sham-operation,model,knockout and overexpression groups were 0,0,(9.60±1.17),(15.20±1.55)and(12.00±1.33)points;the escape latency period at day 7 was(16.15±2.48),(16.98±2.35),(40.72±5.42),(75.42±8.59)and(47.23±6.04)s;the number of positive astrocytes was(1 264.60±135.45),(1 289.20±132.29),(3 269.10±189.39),(103.90±11.09)and(5 301.50±236.29)cell·mm-2;the expression levels of GFAP gene in control,sham-operation,model and overexpression groups were 0.86±0.02,0.92±0.04,1.37±0.07 and 3.42±0.07.Compared with overexpression group,the differences of above indexes in control,sham-operation,model,knockout groups were statistically significant(all P＜0.05).Conclusion TBI causes cognitive impairment and affects cognition and memory in rats,and the GFAP gene is associated with the activation of astrocytes and helps repair nerve damage.

Objective To explore astrocyte activation's impact on cognitive deficits post-traumatic brain injury(TBI).Methods SPF male rats were divided into control,sham-operation and model groups with 10 rats per group.Control group did not receive the surgical intervention;sham-operation group underwent craniotomy without damaging the dura mater;model group was struck with brain injury instrument after craniotomy.The rats with knockout overexpressed glial fibrillary acidic protein(GF AP)gene were set as the knockout group and overexpression group,with 10 rats per group.The knockout and overexpression groups were treated as the model group.Neurological function was evaluated by modified neurological severity score(mNSS).Escape latency was assessed with the Morris water maze test.The expression levels of GFAP in astrocytes were measured quantitative real-time polymerase chain reaction.The positive expression of astrocytes was determined by immunohistochemistry.Results The mNSS of control,sham-operation,model,knockout and overexpression groups were 0,0,(9.60±1.17),(15.20±1.55)and(12.00±1.33)points;the escape latency period at day 7 was(16.15±2.48),(16.98±2.35),(40.72±5.42),(75.42±8.59)and(47.23±6.04)s;the number of positive astrocytes was(1 264.60±135.45),(1 289.20±132.29),(3 269.10±189.39),(103.90±11.09)and(5 301.50±236.29)cell·mm-2;the expression levels of GFAP gene in control,sham-operation,model and overexpression groups were 0.86±0.02,0.92±0.04,1.37±0.07 and 3.42±0.07.Compared with overexpression group,the differences of above indexes in control,sham-operation,model,knockout groups were statistically significant(all P＜0.05).Conclusion TBI causes cognitive impairment and affects cognition and memory in rats,and the GFAP gene is associated with the activation of astrocytes and helps repair nerve damage.

Objective: To understand the characteristics relating to the etiology and complications of hand, foot and mouth disease (HFMD) based on data from the pilot National Sentinel Surveillance (NSS) program so as to explore the feasibility, advantages and disadvantages of the NSS. Methods: Data were extracted from the NSS system, conducted in 11 provinces of China from November 2015 to October 2016. Characteristics regarding the etiology, complications of HFMD and factors related to the positive rates of HFMD specimens were analyzed under the logistic regression method by SPSS 20.0 software. Results: A total of 4 783 specimens were collected, including 3 390 from mild, 1 390 from severe and 3 from death cases. The overall positive rate was 81.43% (3 895/4 783). Other enteroviruses (non EV71/Cox A16 enteroviruses) appeared the major serotype (52.68%, 1 482/2 813) for mild infection of the disease while EV71 was for the severe cases (65.31%, 706/1 081). The serotype spectrum revealed by the pilot NSS was almost identical with the existing surveillance system. Other enteroviruses tended to infect younger children (χ²=130.17, P<0.001) than EV71 and Cox A16, in China. The multivariate logistic regression results showed that higher positive rate was associated with specimens which were collected from males, at children’ hospitals, in peak seasons, timely and in stools. The positive rates presented downwarding trends with the extension of the onset-sampling interval (χ²=14.47, P<0.001 in stool specimen; χ²=31.99, P<0.001 in throat swab; χ²=24.26, P<0.001 in anal swab). Aseptic meningitis, non-brainstem encephalitis and brainstem encephalitis appeared the top three complications of both EV71-associated and other enteroviruses-associated severe HFMD cases. Conclusions Factors as gender, season/place/timeliness of specimen collection, and types of hospital all appeared independently influenced the positive rates. NSS seemed feasible to be used as an alternative or supplement tool to the existing surveillance program in China.

Objective To investigate the clinical severity,etiological classification and risk factors of severe cases with hand,foot and mouth disease (HFMD).Methods A total of 1 489 records on severe and fatal HFMD cases reported to the national pilot surveillance system of HFMD were used to analyze the demographic,medical treatment,etiological classification of the cases.Treatment outcome related risk factors were also studied with multi-variable stepwise logistic regression method.Results Seven out of the 1 489 severe HFMD cases died of this disease.A total of 960 (72.9％) were under three years old and 62.9％ were male and most of the cases (937,62.9％) resided in rural areas.Among all the cases,494 (33.2％) went to seek the first medical assistance at the institutions of village or township level.Durations between disease onset and first medical attendance,being diagnosed as the disease or diagnosed as severe cases were 0(0-1) d,1 (0-2) d and 2 (1-4) d,respectively.In total,773 (51.9％) of the severe HFMD cases were diagnosed as with aseptic meningitis,260 (17.5％) with brainstem encephalitis,377 (25.3 ％) with non-brainstem encephalitis,6 (0.4％) with encephalomyelitis,1 (0.1％) with acute flaccid paralysis,4 (0.3％) with pulmonary hemorrhage/pulmonary edema and 68 (4.6％) with cardiopulmonary failure.Of the etiologically diagnosed 1 217 severe and fatal HFMD cases,642 (52.8％) were with EV71,other enterovirus 261 (21.5％),Cox A16 36 (3.0％),1 (0.1％) with both EV71 and Cox A16.However,277 (22.8％) showed negative on any pathogenic virus.Complication (Z=3.15,P=0.002) and duration between onset and diagnosed as severe cases (Z=3.95,P＜0.001) were shown as key factors related to treatment outcomes.Conclusions Most severe HFMD cases appeared in boys,especially living in the rural areas.Frequently seen complications would include aseptic meningitis,non-brainstem encephalitis and brainstem encephalitis.EV71 was the dominant etiology for severe and fatal cases.Early diagnosis and complication control were crucial,related to the treatment outcome of HFMD.

Objective To explore the assessment methods of dysphagia.Methods The data of 37patients with dysphagia were retrospectively analyzed.These patients took the Kubota drinking test,Tengdao's evaluation,videofluoroscopic swallowing study(VFSS)and fibroptic endoscopic evaluation of swallowing (FEES).Results Fourteen out of thirty-seventh patients showed abnormal results during Kubota drinking test.Tengdao's evaluation results showed that 29/37 patients were abnormal.There 27/37and 33/37 patients showed abnormalities in positive-aspiration score and swallow dysfunction score of VFSS.The number of abnormal patients in aspiration score of FEES was 19/21.The Kappa values were 0.137,0.416 between Kubota drinking test.Tengdao's evaluation and VFSS.The FEES was measured against the VFSS for sensibility,specificity,positive predictive value,and negative predictive value,the values were 88.9％,66.7％,94.1％ and 50.0％.Conclusions Kubota drinking test and Tengdao's evaluation can be applied for screening purpose and evaluating result after treatment;VFSS and FEES can be used as more accurate assessments,they can study the dysphagia's character,position and severity.The combination of a variety of dysphagia evaluation methods is the most important basis for diagnosis and treatment of deglutition disorders.

Objective To explore the feasibility and advantage of fluoroscope in identification of brain structures in nude mice with green fluorescent protein (GFP) expression. Methods We laid the whole brain separated from 8-week adult nude mice with GFP expression into SLY mouse brain blocker to produce slices of 1 or 0.9 mm thickness; and then,25 μm-thickness frozen sections were cut.Fluoroscope was employed to observe the morphological structure to define their anatomic structures with reference to The Mouse Brain in Stereotaxic Coordinates compiled by Paxinos. After the observation,these frozen sections were performed Nissi staining for contrast. Results Different structures can be identified by their distinct fluorescence intensity:the dense areas of nuclei,Nissl bodies and nerve tract showed low fluorescence intensity; while the structures around the areas of nuclei and nerve tract,such as,the plexiform layer of olfactory bulb and the molecular layer of cerebella,showed high fluorescence intensity.The fluorescence intensity was attenuated obviously after Nissl staining; the visualized structural information observed under stereomicroscope was in accordance with that viewed by fluoroscope.Conclusion The identification of brain structure in nude mice with GFP by fluoroscope can serve as an experimental platform being applied in the anatomic structure positioning in fluorescence tracer experiments.

Objective To determine whether the combination of traditional risk factors and quantitative coronary angiography (QCA) assessment could provide accurate prognostic information on a population-based study including 1137 adults with subclinical artherosclerosis and with coronary risk factors. Methods Participants underwent coronary angiography examination before the minimal stenotic diameters, segment diameters, percent stenosis, plaque areas. Other parameters were analyzed by the computer-assisted Coronary Angiography Analysis System. The Framingham Risk Score for each participant was assessed. During the 1 year follow-up period, all kinds of endpoint cardiovascular events were screened. Endpoint events were defined as death from coronary heart disease, nonfatal myocardial infarction (MI) or unstable angina pectoris. Results During the 1 year of follow-up period, a total of 124 participants developed an endpoint event, which was significantly associated with the Framingham Risk Score, calcium of plaques and the plaque areas (all Ps＜0.05).The QCA score incorporated with the QCA parameters was related to the endpoint events. The Framingham Risk Score was combined with QCA score through logistic regression for prediction of end-point events. Data from the ROC analysis showed the accuracy of this prediction algorithm was superior to the accuracy when variables themselves were used. The event-free survival rate was inferior to the control group in participates under high risk, when being screened with this prediction algorithm (P＜0.05). Conclusion The risk of cardiovascular attack in subclinical artherosclerosis individual seemed to be associated with the Framingham Risk Score, calcium of plaques and the plaque areas. When the traditional risk factors (the Framingham Risk Score) were combined with QCA, the new method could provide more prognostic information on those adults with subclinical artherosclerosis.

OBJECTIVESevere nasopharyngeal stenosis (NPS) is a rare complication of uvulopalatopharyngoplasty (UPPP) and very difficult to manage. This report presents our successful treatment experience.

METHODSFrom Nov 1997 to Feb 2006, 6 adults patients with NPS secondary to UPPP were treated in Peking Union Hospital. Two cases was grade II stenosis, received surgery of local pharyngeal and soft palate mucosa flap rotation to enlarge nasopharyngeal airway with stenosis; For the remaining 4 cases with more severe NPS (grade III) who had received 1-3 times unsuccessful repair procedures previously, prolonged nasopharyngeal hollow obturators were used for 6 months after stenosis repair surgery.

RESULTSWith 9-48 months follow-up, All cases results were satisfactory. Nasal obstruction symptom was eliminated, NPS corrected, no velopharyngeal insufficiency complication happened. Daytime removable nasopharyngeal hollow stent obturators with palate support device is more comfortable for patients.

CONCLUSIONSLocal flap rotation to enlarge stenosis airway and prolonged use nasopharyngeal hollow obturators are reliable methods of correction NPS following UPPP.

Adult ; Cicatrix ; complications ; surgery ; Humans ; Male ; Middle Aged ; Nasopharyngeal Diseases ; etiology ; surgery ; Otorhinolaryngologic Surgical Procedures ; methods ; Palate, Soft ; surgery ; Pharynx ; surgery ; Reoperation ; Treatment Outcome ; Uvula ; surgery