Objective To establish a model for preparation of tissue-engineered skin grafts with hTERT cells carrying human papillomavirus type 6 (HPV 6) genome in vitro, so as to lay a foundation for studying HPV life cycle. Methods The full-length linear HPV6 genome and plasmid pEGFP-▲EGFP were electrophoretically cotransferred into hTERT cells. After selection using G418 resistance, Southern blotting was performed to determine the viral load of HPV6 in transfected cells. 3T3 J2 trophoblastic cells, type I rat-tail collagen and hTERT cells containing the full-length HPV6 genes (HPV6.hTERT cells)were mixed and cocultured on metal meshes to form skin graft-like structures. Hematoxylin and eosin (HE)staining was performed to observe the structure of formed skin grafts, an immunohistochemical assay to measure the expression of HPV6 L1 protein, and electron microscopy to observe virus particles in the skin grafts. Results The linear HPV6 gene was successfully transferred into hTERT cells, and Southern blotting showed the presence of HPV6 DNA in the transferred hTERT cells. The HPV6.hTERT cells, which were cocultured with 3T3 J2 trophoblastic cells and type I rat-tail collagen, proliferated and differentiated over time, and gradually formed skin grafts giving the appearance of verrucous hyperplasia. HE staining showed that the cocultured HPV6.hTERT cells could form typical stratified structure of skin after 7 days of cultivation, and histopathologic features of HPV infection, including obvious papillomatous hyperplasia, presence of vesicular cells, hyperkeratosis and parakeratosis, could be observed after 21 days. The immunohistochemical assay showed the expression of HPV6 L1 protein in the upper portion of skin grafts, and electron microscopy revealed the presence of HPV6 virus particles in skin grafts. Conclusions The established model for preparation of tissue-engineered skin grafts using HPV 6 genome-carrying cells provides a basis for biological studies of HPV, but its application is limited to some degree.

OBJECTIVETo explore the crew and passengers' comfort on the Shijiazhuang-Taiyuan passenger dedicated line and physical factors, such as air pressure, noise, wind speed.

METHODSComfort investigation of all the crew (n = 244) and passengers (n = 377) on the Shijiazhuang-Taiyuan passenger dedicated line at speed of 250 km/h and 200 km/h and the detection of the air pressure, noise and wind speed were performed in 2011.

RESULTSSignificantly higher ratio of comfortable feeling, lower ratio of seriously discomfortable feeling were observed in crew and passengers at 200 km/h compared with those at 250 km/h (P < 0.05), as well as rapid disappearance of discomfortable feeling in crew (P < 0.05) and significantly higher ratio of lightly discomfortable feeling and lower ratios of tinnitus and eardrum discomfort induced by air pressure and noise in passengers at 200 km/h. No significant difference was observed in ear discomfort induced by air pressure and noise among crew, and the duration of disappearance of discomfortable feeling among passengers between 200 km/h and 250 km/h. The noise in carriages exceeded the related standard when the high-speed train passing through the tunnels.

CONCLUSIONThe individuals feel more comfortable at 200 km/h than 250 km/h in this line., which may be related with rapid variation of wind speed and noise when the train passes through the tunnels with high speed.

Adolescent ; Adult ; Atmospheric Pressure ; Female ; Humans ; Male ; Noise, Occupational ; Noise, Transportation ; Occupational Health ; Railroads ; Sensation ; Surveys and Questionnaires ; Wind ; Workplace ; Young Adult

OBJECTIVETo study the effects of meconium-stained amniotic fluid on the cord blood IgE level in neonates.

METHODSA total of 404 neonates with meconium-stained amniotic fluid who were born by cesarean delivery between August 2003 and August 2005 (meconium-stained group) and 256 neonates with normal amniotic fluid delivered by cesarean (control group) were enrolled in this study. The meconium-stained group consisted of 80 cases of mild, 62 cases of moderate and 262 cases of severe meconium-stained amniotic fluid. The cord blood IgE level was measured using ELISA.

RESULTSThe cord blood IgE level in the meconium-stained group was statistically higher than that in the control group (t = 4.03, P < 0.01). There were significant differences between the mild and severe meconium-stained subgroups and the control group for the cord blood IgE level (F=4.28, P < 0.01). The cord blood IgE level in neonates with premature rupture of the membrane between the meconium-stained and the control groups was statistically different. Sexes, gestational age, birth weight and birth order were not associated with the IgE level of cord blood.

CONCLUSIONSThe cord blood IgE levels in neonates with meconium-stained amniotic fluid increase. Premature rupture of the membranes may be associated with an increase of cord blood IgE level.

Amniotic Fluid ; Asthma ; etiology ; Fetal Blood ; chemistry ; Humans ; Immunoglobulin E ; blood ; Infant, Newborn ; immunology ; Meconium ; Meconium Aspiration Syndrome

Objective To study the video-electroencephalography (vEEG) background activities of very premature infants within the first 7 days after birth.Method The inclusion criteria were gestational age (GA) within 31 weeks and bedside vEEG monitoring within first 7 days of birth.Those with severe cranial ultrasound (CUS) findings,refused treatment,dead and with seizures on vEEG were excluded.The clinical information of included cases were reviewed and the continuity and sleep-wake cycle (SWC) on their EEG background activities were analysed.Result From June 2017 to April 2018,a total of 56 cases were included.Their GA were (29.4 ± 1.6) weeks,and birth weight were (1 306 ± 316) g.A total of 31 cases were diagnosed with asphyxia(1 min Apgar score less than 8),and no one with 5 min Apgar score below 5.Their umbilical artery blood pHs were 7.29 ± 0.07.Their first CUS and the CUS at corrected gestational age (cGA) full term were all within the normal range.The Gesell neurological assessment at cGA 6 months were all within the normal range.As the cGA increased,the continuity of EEG background activities became more continuous as the normal continuous/discontinuous pattern.The 95％ confidence interval of max interburst interval was 95 s.The SWC first occurred at GA 26 w,but was immature,then became mature as the cGA increased.Conclusion The EEG background activity analysis of relatively normal very premature infants within first week shows that interburst interval can be longer than the previous report,and the SWC first appear at GA 26 w.

Objective To evaluate the feasibility of using amplitude-integrated electroencephalogram (aEEG) to identify epileptic seizures by physicians and nurses in pediatric intensive care unit (PICU) independently.Method Six testees (two PICU physicians versus one EEG physician and two PICU nurses versus one EEG technician) accepted a short-term training,then interpreted aEEG in a single blinded way.These aEEG recordings with synchronous VEEG monitoring were done from January 2013 to May 2015 in PICU.The testees should recognize and mark both the seizure type and the seizure duration from the two-channel recorder (C3/C4) of aEEG (short-term seizure or status epilepticus (SE)).Using raw VEEG monitoring as a gold standard to determine a seizure,the accuracy,missing and error rate of each testees were confirmed,and the reasons of the latter two situations were analyzed by rank sum test and intertestee agreement (k).Result Eighty-two aEEG recordings from 56 patients were interpreted.Thirty-two recordings had 141 epileptic seizures confirmed by VEEG,including 119 short-term seizures and 22 SE.There were 50 recordings without seizure.As for the short-term seizures,the average accuracy of 6 testees by aEEG alone was (66 ±4)％.The accuracy for SE was 100％ in three testees and 95％ in the other three.Missing rate of the seizures were 24.1％-32.6％ in all 6 testees.Those missed seizures were all short-term (duration less than 20 seconds) but one SE.The average error rate was (19 ± 9) times (P =0.000).These false interpretations were misunderstanding,many kinds of artifacts were regarded as epileptic seizures.The accuracy and missing rate among the testees had no significance(P =0.930,0.996),but the error rate had(P =0.000).The inter-testee agreement (k) between two physicians in PICU and the EEG doctor were 0.700 and 0.687 respectively (P ＜ 0.01),which is good.As for two nurses and the EEG technician,the inter-testee agreement (k) was 0.705 and 0.396 respectively (P ＜ 0.01).Conclusion Most of the seizures especially status epilepticus can be detected by PICU staff after short term training.The accuracy of identification of epileptic seizures was similar among observers from PICU and EEG,although some short-term seizures may be missed,and artifacts are mistaken.It's necessary to communicate with EEG doctors and compare with the row VEEG when physicians in PICU find suspicious events.

OBJECTIVETo observe the efficacy of ursodeoxycholic acid (UDCA) combined with Tongdan: Decoction () on immunological indices and histopathological changes in patients with primary biliary cirrhosis (PBC) of IIor III histological stage.

METHODSSixty PBC patients were assigned randomly and equally: to the control group treated with UDCA alone and the treatment group treated with UDCA combined with Tongdan Decoction. The immunological indices and histopathological changes were detected before and after 24-week treatment, and the follow-up lasted for 1-3 years.

RESULTSAfter 24-week treatment, CD4(+)CD28(-) in the peripheral blood was lowered and CD4(+)CD25(+) was increased in both groups, and better effect was shown in the treatment group (P<0.01). The levels of IgM, IgG, and IgA decreased markedly after 96-week treatment in the treatment group (P< 0.05, P< 0.01), while in the control group, only the latter two showed significant decrease after 148 week (all P<0.05). At the end of the 3-year follow-up, the medians of histopathological

CONCLUSIONCombined therapy of Tongdan Decoction and UDCA showed a better therapeutic effect: than UDCA monotherapy on PBC, especially in improving immunological indices and histopathological hepatic changes.

Antigens, CD ; blood ; Biomarkers ; Drug Therapy, Combination ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Immunoglobulin G ; blood ; Inflammation ; blood ; complications ; Liver Cirrhosis, Biliary ; blood ; drug therapy ; immunology ; pathology ; Male ; Middle Aged ; Ursodeoxycholic Acid ; therapeutic use

OBJECTIVETo detect the changes of central venous-to-arterial carbon dioxide difference (P(cv-a)CO(2)) during early goal-directed therapy (EGDT) in patients with septic shock and evaluate its' value in predicting adequate resuscitation and prognosis.

METHODSFrom April 2009 to October 2010, 26 septic shock patients were enrolled in the study. EGDT was performed in all the patients immediately after enrollment. According to the whether they achieved early goal with in the 6 hour or not, patients were separated to EGDT achievement and un-achievement groups. At the onset and after the 6 hours EGDT, mean arterial pressure (MAP), cardiac index (CI), central venous oxygen saturation (ScvO(2)), oxygen delivery (DO(2)), oxygen consumption (VO(2)), oxygen extraction ratio (O(2) ext), lactate, P(cv-a)CO(2) were recorded. The Acute Physiology and Chronic Health Evaluation II (APACHE II) score and 28 day mortality were compared between 2 groups.

RESULTSThere were no significant difference of age and sex between the 15 patients who achieved early goals and 11 patients who did not. EGDT un-achievement patients had higher APACHE II score (21 ± 5) and 28 day mortality (9/11) when compared with EGDT achievement patients (t = 2.985, χ(2) = 4.547, P < 0.05). In EGDT un-achievement group, MAP, CI, DO(2), VO(2), O(2)ext, ScvO(2), Lac, P(cv-a)CO(2) were comparable between the onset and 6 hours after EGDT. However, in EGDT achievement group, MAP ((90 ± 9) mmHg (1 mmHg = 0.133 kPa)), CI ((4.0 ± 1.8) L×min(-1)×m(-2)), DO(2) ((596 ± 274) ml×min(-1)×m(-2)), ScvO(2) (76.9% ± 4.1%) increased, and P(cv-a)CO(2) ((4.2 ± 2.7) mmHg) decreased significantly after 6 hours of EGDT (t values were -3.393, -2.985, -2.103 and -3.195 respectively, all P < 0.05). The changes of P(cv-a)CO(2) between the onset and 6 hours after EGDT, demonstrated high value for predictability of outcome, according to the area under the ROC curve (AUC) was 0.839 (P = 0.004). As a predictor for death, increasing of P(cv-a)CO(2) after 6 hours of EGDT has a sensibility of 100% and specificity of 60%.

CONCLUSIONSIncreasing of P(cv-a)CO(2) after EGDT purports inadequate tissue perfusion in patients with septic shock. Changes of P(cv-a)CO(2) during EGDT demonstrated a useful tool to evaluate adequate resuscitation and prognosis.

Aged ; Aged, 80 and over ; Blood Gas Analysis ; Carbon Dioxide ; blood ; Female ; Humans ; Male ; Middle Aged ; Prognosis ; Resuscitation ; Shock, Septic ; blood ; therapy

BACKGROUNDSevere acute respiratory syndrome coronavirus (SARS-CoV) is a newly emerging virus that gives rise to SARS patients with high rates of infectivity and fatality. To study the humoral immune responses to SARS-CoV, the authors evaluated IgG and IgM specific antibodies in patients' sera.

METHODSTwo methods, enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescent assay (IFA), were used to detect specific serum IgG and IgM against SARS-CoV in 98 SARS patients and 250 controls consisting of patients with pneumonia, health-care professionals and healthy subjects. The serum antibody profiles were investigated at different times over one and a half years in 18 of the SARS patients.

RESULTSThe sensitivity and specificity of ELISA for detecting IgG against SARS-CoV were 100.0% and 97.2% and for IgM 89.8% and 97.6% respectively; the figures using IFA for IgG were 100.0% and 100.0% and for IgM 81.8% and 100.0% respectively. During the first seven days of the antibodies trace test, no IgG and IgM were detected, but on day 15, IgG response increased dramatically, reaching a peak on day 60, remaining high up to day 180 and decreasing gradually until day 540. On day 15, IgM was detected, rapidly reached a peak, then declined gradually until day 180 when IgM was undetectable.

CONCLUSIONThe detection of antibodies against SARS virus is helpful in the clinical diagnosis of SARS.

Adult ; Aged ; Antibodies, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Fluorescent Antibody Technique, Indirect ; Humans ; Immunoglobulin G ; blood ; Immunoglobulin M ; blood ; Male ; Middle Aged ; SARS Virus ; immunology ; Severe Acute Respiratory Syndrome ; immunology ; T-Lymphocytes, Cytotoxic ; immunology

OBJECTIVETo evaluate the correlation between the lactate in artery blood and lactate clearance rate and prognosis in patients with septic shock in intensive care unit (ICU).

METHODSProspectively studied 221 consecutive patients with septic shock from December 2005 to December 2007, the diagnosis of septic shock was made based on the criteria of 2001 ACCP/SCCM. For inclusion in the study, we required admission of the patients within 24 h of septic shock diagnosed. The criteria for exclusion from the study were an age of less than 18 years, pregnancy, serious inadequacy of liver and renal, needing blood purification, or acidosis result of biguanides drugs, or do-not-resuscitate. The eligible patients assigned to early goal-directed therapy. The 6-, 24- and 72-hour lactate clearance rate were calculated, the relationship between the level of lactate, lactate clearance rate, the APACHE II score, the number of failed organ and the 28-day mortality were evaluated.

RESULTSOne hundred and five patients with septic shock were admitted, 74 male and 31 female, the mean age was 70 +/- 12 years. The 28-day mortality was 54.3%. The average APACHE II score at baseline was 20 +/- 8, the number of failed organs was 3.0 +/- 1.1 and the average concentration of lactate in artery blood at baseline was (3.8 +/- 3.6) mmol/L. Significant differences of the lactate at 0-, 6-, 24- and 72-hour were found between death group and survival group. There were 69 patients whose lactate in artery blood at baseline was > 2 mmol/L, 24 survived. The lactate clearance rate of 6- and 24-hour in survival group were significantly higher than death group (P < 0.01, P < 0.05, respectively), but the lactate clearance rate of 72-hour was not (P > 0.05). By using a multivariate logistic regression analysis, it showed that the lactate clearance rate of 6-hour was the independent predictive factor of survival. The area under the receiver operating characteristic curve (ROC) was 0.564, 0.649, 0.754, 0.784, respectively according to the level of the lactate at 0-, 6-, 24-hour and the 6-hour lactate clearance rate. The cutoff of 6-hour lactate clearance rate was >or= 30.0%, resulting in a sensitivity of 60.0% and a specificity of 77.3%.

CONCLUSIONSDynamic observation of lactate level is very important for the survival in the patients with septic shock. Patients with elevated lactate and not decreased rapidly have a worse outcome. The 6-hour lactate clearance rate might be the indicator for predicting the prognosis of patients with septic shock.

Adult ; Aged ; Aged, 80 and over ; Female ; Humans ; Lactic Acid ; blood ; Male ; Middle Aged ; Monitoring, Ambulatory ; Prognosis ; Prospective Studies ; Shock, Septic ; blood ; Young Adult

OBJECTIVETo investigate the estrogen receptors (ER)alpha and ERbeta expression and their relationship with clinicopathological parameters in human breast carcinoma.

METHODSSamples were obtained from 30 breast carcinoma, reverse transcriptase polymerase chain reaction was used to measure the expression of ERalpha and ERbeta mRNA.

RESULTSERalpha mRNA level was up-regulated in breast carcinoma tissue compared with adjacent normal tissue (t = 7.399, P < 0.01) while down-regulated in ERbeta. The relative ratio of ERalpha and ERbeta was decreased in normal tissue vs. carcinoma (t = 6.385, P < 0.01), in patients with lymph node metastasis vs. those without lymph node metastasis (t = 2.602, P < 0.05), in late stage carcinoma vs. early stage (t = 3.754, P < 0.05).

CONCLUSIONERalpha and ERbeta play divergent role in the development of human breast carcinoma.

Adult ; Aged ; Aged, 80 and over ; Breast Neoplasms ; metabolism ; pathology ; physiopathology ; Cell Line, Tumor ; Estrogen Receptor alpha ; biosynthesis ; genetics ; physiology ; Estrogen Receptor beta ; biosynthesis ; genetics ; physiology ; Female ; Humans ; Middle Aged ; RNA, Messenger ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction