Objective:To explore the pathogenic spectrum and epidemiological characteristics of respiratory viruses in children with acute lower respiratory tract infection in Changsha, and provide scientific basis for disease prevention and treatment.Methods:A total of 1 092 respiratory tract specimens of children were collected and 12 respiratory viruses were detected by real-time quantitative transcription polymerase chain reaction.Results:Among the samples from 1 092 cases, those from 437 cases (40%) were positive for respiratory syncytial virus (RSV), 337 cases (30.9%) were positive for parainfluenza virus-3 (PIV-3), 263 cases (24.1%) were positive for human bocavirus (HBOV) and 228 cases (20.8%) were positive for adenovirus (ADV). The detection rates of boys and girls were 82.26% and 83.42%. The infection rate of RSV was higher in the group ≤ 6 months of age, the infection rates of PIV-3 and HBOV ≤2 years old were higher, and the infection rate of ADV was higher in the group between 6 months to 5 years old. The detection rates of virus infection of spring, summer, autumn and winter were 90.48%, 83.50%, 62.26% and 82.80%, respectively, there were significant differences among them.Conclusions:The main viruses in children with acute lower respiratory tract infection in Changsha were RSV and PIV-3. Mixed infections were common. Children under 2 years of age were more likely to get acute lower respiratory infections. Viruses had seasonal trends and peaked in winter and spring.

Objective: To analyze the clinical characteristics of pheochromocytoma crisis (PCC) . Methods: Data of 123 cases of pheochromocytoma and paraganglioma (PPGL) admitted from Apr. 2011 to Feb. 2017 were retrospectively analyzed and they were divided into crisis group and noncrisis group according to the patients with or without haemodynamic instability and end-organ damage. The differences of demographics characteristics, presentations, laboratory tests, imaging findings, perioperative clinical conditions and pathological features were compared between the two groups. Results: ①16 cases were enrolled into crisis group, among whom 5 were misdiagnosed, while 107 cases were enrolled into noncrisis group. ②Compared with noncrisis group, the incidence of headache, palpitation, sweating, the classic triad, other presentations of PPGL, severe hypertension and hypotension were higher, and more patients had paroxysmal hypertension and admitted to our hospital for paroxysmal presentations in crisis group (P<0.05) . ③Leukocyte, fasting blood glucose, liver transaminases, troponin and D-dimmer were higher, while estimated glomerular filtration rate (eGFR) was lower, more tumors located in the left of adrenal in crisis group (P<0.05) . ④ Patients in crisis group had higher plasma free metanephrines (MNs) , larger maximal tumor diameter, higher enhanced CT value in each period, more benign tumors and hemorrhage or necrosis in the tumors, but all the differences were not significant when compared with the noncrisis group. ⑤Patients in crisis group were more likely to undergo elective surgery. However, there was no difference in the preoperative time of α-blockade, type of surgery, intraoperative and postoperative complications, mortality among the two groups. Conclusions PCC is a rare endocrinological emergency with a highly variable manifestations, which commonly presents with typical triad, with higher incidence of hemodynamic instability and end-organ damage. Although biochemical and imaging examinations are relatively effective in the diagnosis of PCC, the misdiagnosis remains inappropriate high. Once the diagnosis is established, clinicians should timely start drug preparation while surgical resection is the key to the treatment of PCC.

Objective: To investigate the incidence of occult cervical lymph node metastasis and the common neck level of metastases in cN0 laryngocarcinoma, and the relationship between the clinicopathologic features of laryngocarcinoma and cervical lymph node metastasis. Methods: A total of 506 cases with cN0 laryngocarcinoma treated at the First Affiliated Hospital of Chongqing Medical University between March 2011 and March 2018 were enrolled, and their medical records and follow-up data were retrospectively analyzed. Of them, 211 cases of were glottic carcinoma in stage T1 without neck dissection and they were observed by clinical follow-up; other 295 cases, including glottic carcinoma, supraglottic carcinoma and hypopharyngeal carcinoma in stage T2-T4 were treated with surgical resection of the primary lesions and selective neck dissection. SPSS 22.0 software was used to analyze the data. Results: The total incidence of cervical lymph node metastasis was 10.87%(55/506), with a lower incidence in T1 stage glottic carcinoma(6/211,2.84%) than that in other cases(49/295,16.61%). The incidence of cervical lymph node metastasis in glottic carcinoma (29/426, 6.81%) was lower than those in supraglottic carcinoma (22/71,30.99%) and subglottic carcinoma (4/9) (χ²=35.810,P<0.01).The pN+ rates of glottic carcinoma at T1, T2, T3 were 2.84%(6/211), 5.31%(6/113), 16.05%(13/81), and 19.05%(4/21), respectively (χ²=18.572, P<0.01). The pN+ rates of supraglottic carcinoma at T2, T3 and T4 were 3/13, 32.50%(13/40) and 6/13, respectively (χ²=3.649,P>0.05). The incidence of cervical lymph node metastasis in poorly differentiated carcinoma (17/42, 40.48%) was higher than those in moderately differentiated carcinoma (26/205, 12.68%) and high differentiated carcinoma(12/246, 4.88%)(χ²=36.356, P<0.01). Moreover, 85 pN+ lymph nodes were obtained by selective neck dissection, respectively 43(50.59%) in level Ⅱa, 30(35.29%) in level Ⅲ, 1(1.18%) in level Ⅳ and 11(12.94%) in level Ⅵ. Conclusions The occult cervical lymph node metastasis was frequently found in cN0 laryngocarcinoma. Selective neck dissection should be performed with surgery for the primary lesions in T3-T4 glottic laryngeal cancer, T2-T4 supraglottic laryngeal cancer and subglottic carcinoma, and the neck dissection for level Ⅱa and Ⅲ is appropriate. It is required to detect pre-laryngeal and pre-tracheal lymph nodes in patients with subglottic laryngeal carcinoma.

Objective To investigate the value of serum IgG4 level for the diagnosis of IgG4-related hepatobiliary diseases and the differentiation from other hepatobiliary diseases.Methods A total of 270 patients with hepatobiliary diseases in the People's Hospital of Hunan Province from August 2015 to April 2017 were enrolled in this study,and 20 healthy subjects were selected as controls.The 270patients were divided into eight groups:liver cirrhosis group (n =17),acute pancreatitis group (n =52),chronic pancreatitis group (n =33),cholecystitis and gallstone group (n =27),bile duct carcinoma group (n =30),cholangitis and biliary calculi group (n =41),pancreatic cancer group (n =47),IgG4-related hepatobiliary disease group (n =23).The levels of serum IgG4 were measured by rate nephelometery assay.The sensitivity and specificity of IgG4 levels for distinguishing IgG4-associated hepatobiliary diseases were evaluated by receiver operating characteristic curve.Results The levels of IgG4 of the cirrhosis group and the IgG4 related hepatobiliary disease group were significantly higher than those of the control group (P ＜ 0.05).The IgG4 level in the hepatobiliary disease group was significantly higher than those of the other seven groups (Z =-5.267,-6.802,-5.921,-6.005,-6.173,-6.513,-6.014,P all ＜ 0.01).The area under curve (AUC) for IgG4 level in distinguishing IgG4 associated hepatobiliary diseases and other hepatobiliary diseases was 0.982.When 4.13 g/L was used as the cut off value of diagnosis,the sensitivity and specificity of IgG4for diagnosis were 95.7％ and 96.0％ respectively.The IgG4 levels in twelve patients with IgG-associated hepatobiliary diseases after 2 months of glucocorticoid therapy were significantly lower than those before glucocorticoid therapy (Z =-2.021,P =0.043).Conclusion The elevated serum IgG4 level may not be specific just for IgG4-related hepatobiliary diseases.The cut off value of 4.13 g/L should be very useful for diagnosing IgG4-related hepatobiliary diseases,differentiating from other hepatobiliary diseases and evaluating the therapeutic effect of glucocorticoid therapy.The further detailed verification for these findings should be necessary in clinical practice by increasing the sample size.

Objective To observe the levels of interferon gamma-induced protein 10(IP-10)in children with acute viral and bacterial infection.Methods There were three groups:acute viral infection group(51 ca-ses),acute bacterial infection group(52 cases),and healthy control group(51 cases).Serum IP-10 levels were detected by enzyme linked immunosorbent assay(ELISA),and serum C reactive protein(CRP)were deter-mined with BNⅡ automatic protein analyzer.Results The level of serum IP-10 and CRP were significantly different in different groups(P<0.05).Compared with control group,the level of IP-10 and CRP were higher in acute viral infection group or in acute bacterial infection group(P<0.05).The level of IP-10 was higher in acute viral infection group than that in acute bacterial infection group(P>0.05).The level of CRP was higher in bacterial infection group than that in acute viral infection group(P<0.05).The IP-10 levels in viral Infec-tion patients were not correlated with the CRP levels.The area under curve(AUC),sensitivity and specificity of IP-10 and CRP were 0.688 and 0.873,35.0% and 79.6%,96.1% and 98.0%.When cut-off value of predic-tive probability was 0.713,sensitivity and specificity were increased to 82.5% and 100.0%.Conclusion Ser-um IP-10,CRP and predictive probability are valuable in the diagnosis of acute pathogen infection in children.

Objective To explore the cause constituents of neonatal severe hyperbilirubinemia and the clinical efficacy and safety of blood exchange transfusion treatment .Methods 142 neonates with severe hyperbilirubinemia conducted the blood exchange transfusion therapy .The levels of serum total bilirubin ,indirect bilirubin and direct bilirubin and the change of blood routine indica-tors were analyzed before and after blood exchange transfusion .Results The main causes leading to neonatal severe hyperbilirubi-nemia were bacterial infection(28 .20% ) ,glucose-6-phosphate dehydrogenase(G6PD) deficiency(27 .50% ) and pregnant women with ABO blood group incompatibility (16 .20% ) .The levels of serum total bilirubin ,indirect bilirubin ,direct bilirubin and blood routine indicators after operation in neonates with severe hyperbilirubinemia were significantly lower than those before operation , the differences were statistically significant (P<0 .05) .The total bilirubin swap exchange was (54 .40 ± 9 .90)% .The intraoperative adverse reactions rate was 3 .50% .The postoperative thrombocytopenia occurrence rate was 72 .00% .Conclusion The blood ex-change transfusion for treating neonatal severe hyperbilirubinemia possesses has clinical significance ,but the hematology and bio-chemical indicators monitoring should be strengthened for avoiding adverse reactions occurrence .

Objective:To study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs.Methods:BABL/c mice were randomly divided into VLPs experimental group, alum adjuvant experimental group, PBS control group and alum adjuvant control group,the experimental group mice were intramuscular immunization with HBoV1 VP2 VLPs and HBoV1 VP2 VLPs added alum,control group mice were immunization with alum or PBS buffer,then to study the effect of alum on immune response in mice induced by HBoV1 VP2 VLPs by cellular and humoral immune strength.Results: Alum adjuvant decreased cellular immune response induced by HBoV1 VP2 VLPs(P<0.001),enhance the HBoV1 VP2 VLPs immuned serum IgG titer(P<0.05)and IgG activity(P<0.01).Conclusion:Alum adjuvant can enhance humoral immune response induced by HBoV1 VP2 VLPs,but weaken cellular immune response induced by HBoV1 VP2 VLPs,when HBoV1 VP2 VLPs used as a prophylactic vaccine it should add alum adjuvant,when used as a therapeutic vaccine,it should not add alum adjuvant.

Objective To establish and evaluate a Metapneumovirus real-time quantitative reverse transcription polymerase chain reaction (RT-PCR),detect clinical specimens and explore the clinical prevalence characteristics of Metapneumovirus-infected children.Methods The primers and probes which targeted the conserved genes F were designed,RNA standards were prepared to establish a standard curve,the sensitivity,specificity and reproducibility had been tested.222 lower respiratory clinical specimens were collected from children in Lanzhou area with ARI.Metapneumovirus and co-infection viruses were detected simultaneously;further Metapneumovirus related epidemiology was studied.Results Metapneumovirus linear detection range was 10-10s copies/μl,the lowest detection limit was 10 copies/μl,the correlation coefficient was 1,the amplification efficiency was 91.62％,the CV of Ct value was less than 2％.Take conventional PCR product sequence results as reference,real-time quantitative RT-PCR sensitivity and specificity were 100％ and 96.17％.Metapneumovirus detection rate were 9.46％,13 cases for boys (5.86％),8 case for girls (3.60％).The detection rate of spring,summer,autumn and winter were 15.71％,0％,5.08％,11.11％.There were no significant differences between the Metapneumovirus viral load and mixed infection or the types of disease,clinical symptoms.Conclusions Metapneumovirus realtime quantitative RT-PCR has been confirmed as a sensitive and specific method.Metapneumovirus was an important agent of children respiratory tract infection in Lanzhou region.We should take the long-term systematic surveillance seriously.

Objective By comparing the biological characteristics among Lewis lung cancer cells ( LLC) , LLC or-thotopic transplantation derivative cells ( R1-LLC) and R1-LLC orthotopic transplantation derivative cells ( R2-LLC) , we e-valuate their invasion and metastatic abilities in orthotopic transplantation models.Methods In vitro, we applied CCK8, clonogenic assay, and Transwell invasion assay to detect the proliferation ability, invasion ability and morphologic structures of LLC,R1-LLC, R2-LLC cells respectively, meanwhile observing morphologic structures by transmission electron microsco-py.In vivo, we constructed LLC, R1-LLC and R2-LLC orthotopic transplantation models.Herein, we observed their tumor growth and metastasis.The tumor formation rate and tumor-forming time were recorded for statistic analysis.Results In vitro:LLC, R1-LLC and R2-LLC cells showed no significant difference in proliferation ability ( P>0.05 ) , but significant differ-ences in invasion ability:R2-LLC>R1-LLC>LLC(P<0.05).In vivo:In the orthotopic group, the tumor formation rates of LLC, R1-LLC and R2-LLC cells were 66.67%、80%、93.33%(P <0.05).Conclusions In orthotopic implantation mouse models, R2-LLC cells present higher invasion and metastatic ability than R2-LLC and LLC cells.

Objective To express the VP8 * protein of the rotavirus vaccine strain LLR in the E.coli with the pGEX4T-1 vector,which is important for the further research of the VP8 * protein functions.Methods The LLR VP8 * gene was obtained by virus RNA extraction and RT-PCR.Then it was cloned in the expression vectors pGEX4T-1.The recombinant plasmid pGEX4T-1-VP8 * was transformed to the E.coli BL21.Then the VP8 *-GST fusion protein was expressed and purified by the affinity chromatograph.The protein of interest was validated by SDS-PAGE and Western Blot.Results The molecular weight of the VP8 *-GST fusion protein was about 52 000 according to the SDS-PAGE.The bands of both 52 000 and 26 000 were shown in the Western Blot with the antibody against GST.Conclusions The LLR VP8 * gene was obtained and cloned to the pGEX4T-1 vector.Moreover,the solvable VP8 *-GST fusion protein was successfully expressed and purified.