Objective:To analyze the clinical characteristics, examination results, treatment and prognosis of neonates with influenza virus infection in the neonatal intensive care unit (NICU).Methods:Clinical data of neonates with influenza virus infection who were hospitalized in the NICU of the General Hospital of Southern Theater Command of Chinese People′s Liberation Army from January 2018 to December 2020 were retrospectively analyzed.Results:A total of 11 hospitalized neonates with influenza virus infection in the NICU were recruited, including 2 cases of influenza A and 9 cases of influenza B. Ten cases (90.9%) had respiratory symptoms, and among them, there were 8 cases with increased oxygen demand, 7 cases with complicated pneumonia, 4 cases with dyspnea, and 2 cases with apnea.Seven cases showed abnormal body temperature, including 6 cases of fever, and 1 case of hypothermia.Five cases had circulatory system symptoms.Digestive system symptoms and urinary system symptoms were detected in 5 cases and 3 cases, respectively.Eight cases complicated with systemic symptoms, including 3 cases of poor mental response, 3 cases of worsening jaundice, 3 cases of weight loss, 2 cases of hyperglycemia, 1 case of edema and sclerosis.Ten cases were treated with gamma globulin immunotherapy, 2 cases were treated with plasma immune support, and 1 case was treated with Peramivir antiviral.Eight cases were treated with increased oxygen therapy, among which non-invasive ventilator parameters or modes increased in 4 cases, and nasal cannula oxygen concentration increased in 2 cases.The change of noninvasive-assisted ventilation to invasive-assisted ventilation occurred in 1 case, and 1 case developed the change of nasal cannula to noninvasive-assisted ventilation.Four neonates received anti-shock and (or) myocardial contractility therapy.Conclusions:Neonates with influenza virus infection in the NICU are mainly manifested as respiratory symptoms and fever, and the incidence of complicated pneumonia is high.Multiple systems may be involved at the same time, often leading to severe disease status.Comprehensive supportive treatment is necessary.Neonatologists should pay attention to these symptoms, and early detection of influenza virus and timely isolation are the key methods to prevent influenza outbreaks in NICU.

【Objective】 To analyze the risk factors related to the number of RBCs transfusion in neonates with neonatal respiratory distress syndrome (NRDS), and to explore the complications and the predictive indicators related to the increase of RBCs transfusion frequency. 【Methods】 NRDS newborns admitted to our hospital from January 2017 to January 2019 were selected and divided into three groups according to the number of RBCs transfusion, namely, non-transfusion group, 1~ 2 times group, and ≥3 times group. The clinical data and complications of the three groups were compared, and the risk factors leading to the increase of the number of blood transfusion were analyzed. 【Results】 Such factors as maternal age ≥35 years old, gestational age, birth weight, hemoglobin(Hb) at admission, non-invasive ventilation time, hospitalization time in neonatal intensive care unit(NICU), total enteral feeding time affected the blood transfusion frequency of the three groups(χ²=14.24, F=28.44, 41.70, 60.05, 3.83, 5.97, 4.40, P<0.05). The incidence of necrotizing enterocolitis (NEC), septicemia and feeding intolerance in blood transfusion ≥3 times group was significantly higher than that in 1~2 times group and non-transfusion group (χ²=19.30, 18.68, 6.98, P<0.05). Multivariate logistic regression analysis showed that birth weight, Hb at admission, length of stay in NICU and time of reaching total enteral feeding were independent risk factors for≥ 3 times of blood transfusion (OR=-3.942, -0.186, 0.530, 0.324, P<0.05). The ROC curve showed that birth weight and Hb at admission were effective in predicting blood transfusion ≥3 times, and the area under the ROC curve were 0.846 and 0.802, respectively, and the truncation values were 2.315 kg and 157.5 g/L. 【Conclusion】 Feeding intolerance, NEC and septicemia are the complications of the increased transfusion frequency in children with NRDS, and birth weight and hemoglobin at admission are effective in predicting blood transfusion ≥ 3 times.

Objective To observe the effect and quality of life score of laparoscopic choledochotomy and T tube drainage for the patients of extrahepatic bile duct stones.Methods A total of 80 patients with calculus of extrahepatic bile duct in Shandong Provincial Third Hospital from February 2016 to April 2017 were retrospectively analyzed,including 41 males and 39 females,among which,40 patients were treated with laparoscopic choledocholithotomy and T tube drainage(the study group),and the other 40 cases were operated on laparotomy (the control group) by hierarchical randomization grouping.After operation,the time of operation,the amount of bleeding during the operation,the time of postoperative exhaust,the time of postoperative hospitalization and the difference of adverse reactions were observed in the two groups.Follow-up at the end of 1 st and 3th month,quality of life score was compared between the two groups.The measurement data with normal distribution were expressed as mean ± standard deviation (Mean ± SD),comparison between groups were performed using t test.The count data were expressed as rate(％),comparison between groups were performed using chi-square test.Results Comparison of data between the study group and the control group:operation time respectively were (97.23 ± 10.21) min,(117.52 ± 10.01) min,the amount of bleeding respectively were(87.73 ± 10.54) ml,(185.13 ± 11.56) ml,postoperative exhaust time respectively were (17.57± 2.96) h,(38.44 ± 3.06) h and the postoperative hospitalization time were (7.75 ± 1.21) d,(12.03 ± 3.85) d.The data of each group in the study group were lower than those in the control group,and the difference was statistically significant(P ＜ 0.05).The quality of life scores of the 1 months and 3 months after treatment in the study group were (45.82 ± 3.22) scores and (47.29 ± 3.09) scores,the control group were (32.56 ± 3.29) scores and (36.19 ± 3.06) scores.The study group was significantly superior to the control group,and the difference was statistically significant(P ＜ 0.05).The complications of the patients in the study group were not statistically significant between the control group and the control group(P ＞ 0.05).Conclusions Compared with open surgery,laparoscopic choledocholithotomy with T tube drainage has significant clinical effect in the treatment of extrahepatic bile duct stones.It can improve the quality of life and safety,and it is suitable for clinical application.

Objective To analyze the clinical features of patients with sporadic adult-onset neuronal intranuclear inclusion disease (NIID),and raise awareness of the disease among medical workers.Methods The clinical data of two patients with pathologically confirmed adult sporadic NIID,admitted to our hospital in February 2018 and October 2018,were collected.The clinical manifestations,head MR imaging,cutaneous pathological features,treatments and prognoses were retrospectively analyzed.Results Both patients were characterized with slow progressive dementia,accompanied with diverse clinical manifestations involving the central and autonomic nervous systems.Acute encephalopathic signs occurred in both patients.Head MR imaging showed extensive leukoencephalopathy mainly in the frontal and parietal lobes;these white matter abnormalities showed hyperintensity in T2-weighted imaging and liquid attenuated inversion recovery sequences,and equal or low signal in T1-weighted imaging.Remarkably,specific curve-like high-intensity signals along the corticomedullary junction in the bilateral frontal lobe were both observed in diffusion-weighted imaging.Patient 2 with seizures showed unilateral cerebral cortical edema on head MR imaging.In both patients,skin biopsy revealed specific eosinophilic inclusion bodies in the nucleus of some sweat gland cells,adipocytes and fibroblasts.Patient one was treated with dexamethasone intravenous drip to relieve headache and vision loss,and cognitive therapy was given.The acute encephalopathy of patient two was relieved by intravenous gamma globulin.Conclusions NIID has various clinical manifestations of central,peripheral,and autonomic nerve systems.Head MR imaging characteristic changes and skin pathological biopsy contribute to the diagnosis.Immunomodulatory therapy may be effective for acute encephalopathic symptoms in NIID.

Objective: To construct the mmu-miR-155 eukaryotic overexpression vector pmR-155 and to investigate its effect on HBV replication and expression of PTEN in vivo. Methods: The mmu-mir-146a precursor gene fragment pre-mmu-mir-146a was amplified by PCR, then connected to the pmR-mCherry plasmid vector after double enzyme digestion, the accuracy of recombinant vector was verified by colony PCR、double enzyme digestion and sequencing; then the recombinant vector was transfected HBV transgene mice(Experimental Group)with hydrodynamics-based injection via vena caudalis, and pmR-mCherry plasmid、PBS were respectively transfected into the mice as Empty plasmid Group、Blank Group. The concentration of IFN-γ in the serum was detected by ELISA. The expression of SOCS1、PTEN mRNA in the liver was detected by qPCR at 30d post-transfectioned. The Western blot was performed to detect the changes in SOCS1、PTEN、HBX in the liver tissue at 30 d post-transfectioned. The results were analyzed with Student’s t-test, or one-way analysis of variance and the least significant difference test. Results: the colony PCR、double enzyme digestion and sequencing verified that the gene was inserted into the pmR-mCherry vector. Compared with Blank Group, the expression of miR-155 in the Experimental Group was significantly increased(t = 8.90, P < 0.01); the concentration of IFN-γ in the Experimental Group was significantly increased(F = 26.58, P < 0.01); the mRNA(F_{SOCS1 mRNA} = 19.72, P < 0.01; F_{PTEN mRNA} = 7.38, P < 0.05) and protein(F_SOCS1 = 50.30, P < 0.01; F_PTEN = 129.00, P < 0.01) expression of COCS1、PTEN was significantly decreased in the Experimental group and the protein of HBX was also significantly(F_HBX = 77.97, P < 0.01). Conclusion The pmR-155 eukaryotic overexpression vector is successfully constructed, this recombinant vector can express miR-155 stably; miR-155 can down-regulate cocs1、PTEN gene expression and up-regulate the expression of IFN-γ, it can inhibit the replication of HBV and a potential targets to treating hepatocellular carcinoma.

Objective To compare the clinical efficacy of subinguinal microsurgical and single -port laparoscopic high ligation in the treatment of varicocele.Methods 218 patients with varicocele were enrolled in this study.According to the digital table,they were randomly divided into two group.148 cases were treated by subinguinal microscopic varicocelectomy(microscopic group),70 cases were treated by single -port laparoscopic high ligation varicocele(single -port laparoscopic group).Postoperative follow up was 3 -24months.The operative duration,length of hospital stay,hospitalization expense,postoperative complications and semen quality parameters were compared between the two groups.Results There were statistically significant differences in operative duration and hospitalization expense between the two groups(all P <0.05).There was no statistically significant difference in length of hospital stay (P >0.05).In the 218 followed -up patients,the sperm concentration and motility (grade a + b sperm)all significantly improved,which of the microscopic group and single -port laparoscopic group preoperation were (19.1 ± 8.2)×106 /mL,(18.2 ±7.9)×106 /mL and (22.7 ±7.8)%,(21.6 ±8.9)% respectively,which at 3 -6 months after operation were (56.2 ±10.8)×106 /mL,(45.8 ±12.9)×106 /mL and (58.8 ±9.7)%,(44.6 ±10.7)%, there were statistically significant differences compared with preoperation (t =6.227,9.579,all P <0.05 ). Conclusion The surgical methods in the treatment of varicocele can improve the quality of patients,but microscopic group is obviously better than single -port laparoscopic group in improvement of semen quality parameters,safety, patient -based compliance and economy.

Objective To construct the has-miR 146a eukaryotic overexpression vector pmR 146a and to explore its effect on the expression of c-Myc gene in HepG2.2.15 cells.Methods The has-miR-146a precursor gene fragment pre-has-miR-146a was amplified by PCR,then connected to the pmR-mCherry plasmid vector after double enzyme digestion,the accuracy of recombinant vector was verified by colony PCR,double enzyme digestion and sequencing;then the recombinant vector was transfected into HepG2.2.15 cells as the experimental group,meanwhile the empty vector group (transfecting pmR-mCherry empty plasmid group) and blank group(transfecting reagent lip2000+PBS),then the fluorescent protein expression amount was observed under the fluorescence microscopy at 24,48 h;the expression of has miR-146a was evaluated by qPCR;at 24,48 h after transfection,the expression levels of c-Myc gene mRNA were detected by qPCR,and the c-Myc protein expression level after 48 h was detected by Western blot.Results The colony PCR,double enzyme digestion and sequencing verified that the pre-has-miR-146a gene fragment was inserted into the pmR-mCherry vector;at 24,48 h after transfection in the experimental group and empty vector group,intracellular strong fluorescence was seen by fluorescent microscope,the transfection efficiency was at 50％-60％ contrasting without fluorescence;the has-miR-146a expression level in the experimental group was significantly higher than that in the empty vector group and blank group (P＜0.01);the c-Myc mRNA expression at 24,48 h after tranfection was significantly lower than that in the empty vector group and blank group (P＜0.05);the protein expression amount at 48 h after transfection was lower than that in the empty vector group and blank group (P＜0.01).Conclusion The pmR-146a eukaryotic overexpression vector is successfully constructed,this recombinant vector can express miR-146a stably;miR-146a can down-regulate c-Myc cancer gene expression,which can serve as one of potential targets for treating hepatocellular carcinoma.

Objective To study the diagnosis,treatment and therapeutic results of 11 patients who suffered from mucinous tumor of the bile duct.Methods Eleven patients who were diagnosed to suffer from mucinous tumor of the bile duct were retrospectively studied.Three patients who presented with obstructive jaundice were diagnosed on ERCP,and 8 patients who had extra-and intrahepatic cholangiolithiasis were diagnosed by biopsy during choledochoscopy.Results One of the 3 patients who underwent ERCP died from obstructive jaundice after failed drainage of bile using endoscopic nasobiliary drainage (ENBD).The remaining two patients underwent laparoscopic common bile duct exploration and T tube drainage.The eight patients who had extra-and intrahepatic cholangiolithiasis were diagnosed by biopsy during choledochoscopy.Conclusions The clinical presentation of mucinous tumor of bile duct is non-specific and the preoperative misdiagnosis rate is high.Common bile duct exploration,T tube drainage and long-term T tube drainage is a good way to treat mucinous tumor of the bile duct.

Objective To construct the miRNA‐21 eukaryotic overexpression vector pmR‐21 and to explore its regulation effect on the expression of c‐myc gene in HepG2 .2 .15 cells .Methods The miRNA‐21 precursor gene fragment pre‐miRNA‐21 was amplified by PCR ,then connected to the pmR‐mCherry plasmid vector after double enzyme digestion ,the accuracy of the recombi‐nant vector was verified by double enzyme digestion and sequencing ;then the recombinant vector was transfected into HepG2 .2 .15 cells ,the fluorescent protein expression was observed under the fluorescence microscopy at 24 h and the transfection efficiency was detected by flow cytometry ;the expression of miRNA‐21 was evaluated by real‐time quantitative PCR;at 72 h after transfection ,the expression levels of c‐myc gene were detected by RT‐PCR and Western blot ;CCK‐8 was used to detect the cell proliferation in each group .Results The double enzyme digestion and Western blot verified that the target gene fragment was inserted into the pmR‐mCherry vector;at 24 h after transfection ,intracellular strong fluorescence was seen ,the transfection efficiency was higher than 50% ;miRNA‐21 expression level of the pmR‐21 recombinant vector group was significantly increased;c‐myc gene expression was increased in the pmR‐21 recombinant vector group at 72 h after transfection ,the cell proliferation in the pmR‐21 recombinant group was faster than that in the control group(P<0 .05) .Conclusion The pmR‐21 eukaryotic overexpression vector is successfully con‐structed ,this recombinant vector can express miRNA‐21 stably ;miRNA‐21 can up‐regulate c‐myc gene expression ,c‐myc gene is one of miR‐21′s targets for playing a cancer‐promoting action .

Objective To observe the clinical efficacy and safety of GINA regimen and GINA regimen combined with oral Huaiqihuang granule in the treatment of children with bronchial asthma.Methods A ran-domized,single blind,multicenter,parallel controlled clinical trial wascarried out.A total of 1 128 patients with bronchial asthma in children were randomized into two groups.The observation group were treated with GINA regimen combined with oral Huaiqihuang granule.The GINA regimen treatment group was treated by GINA reg-imen.Clinical assessment and C-ACT scores was observed in first month,third month,sixth month after treat-ment.Clinical assessment included the times of upper respiratory tract infection occurrence,bronchitis and pneu-monia,asthmatic attacks,application of emergency medicine,hospitalizations due to asthmatic.Drug adverse effect in the two groups was compared.Results The times of upper respiratory tract infection,bronchitis and pneumonia,asthmatic was significantly decreased(P <0.05 ),and C-ACTscores were significantly higher(P <0.05)in the first month,the third month,and the sixth month after treatment.There were 16 cases of drug relat-ed adverse reactions.Seven cases were in observation group(n ﹦7)(1.15%)and 9 cases in the GINA regimen treatment group(n ﹦9)(1.73%).There was no significant difference of adverse effect between the two groups (P >0.05).Conclusion The treatment of bronchial asthma in children with GINA regimen combined with oral Huaiqihuang granule can significantly reduce the incidence of respiratory infections and the number of asthmatic attacks dramatically and safely improve clinical curative effect,asthma control.