Objective:Todevelop the isolation and purification technology for typeⅡalveolar epithelial cells(AECⅡ)of fetal rat,in order to provide experimental means for the study of lung development or neonatal lung disease.Methods:Lung tissues of 19-day fetal rats were digested with trypsin and collagenase,then purified for AECⅡwith different centrifugal force and repeated attachment.Cell viability was evaluated by typran inclusion dying before plated into six-well flask.Growth status and shape of attached cells were observed with inverted phase contrast microscope.AECⅡwere identified by electron microscopy and its percentage was assessed by modified Papanicolaou dying and immunofluorescence,the latter aiming to detect expression of surfactant protein C(SPC)in AECⅡ.Results:The total amount of cells we harvested from 3 to5 fetal rat was(36?5)?106 with a viability of 98%?2%.Cells were like polygonal and connected like island under microscope.AECⅡwas ascertained by lamellar bodies found in cytoplasma under electron microscope and its percentage was 96%?3%identified by modified Papanicolaou dying,the same as the result by immunofluorescence for SPC.Conclusion:Highly Purified and viable AECⅡcould be achieved by our methods and the cell model could be used in further study.

Objective To summarize clinical features and changes of peripheral blood lymphocyte subsets in children with EBV-associated hemophagocytic syndrome. Methods Twenty children diagnosed with EBV-AHS were characterized by hemophagocytic syndrome and had the evidence of EBV infection.Clinical characteristics were analyzed and proportions of lymphocyte subsets of CD4+ ,CD8+, CD19+,CD16+CD56+ and CD4+/CD8+ in peripheral venous blood were determined by flow cytometry. Twenty healthy children matched in age and gender with these patients were selected as control. Results In all the diagnostic items of EBV-AHS, fever, decline of cells over two lineages in blood routine, enhanced lactate dehydrogenase ( ＞1 000 U/L) and ferritin ( ＞ 1 500 g/L),decreased high-density lipoprotein occurred in all the patients with an incidence of 100%. The percentages of CD8 + T and CD19 + B in children with EBV-AHS were higher than those in control group(53.70% ± 10.6% vs 27.05% ±8.22%,24.95% ±5.34% vs 11.85 % ± 4. 53 %, respectively), while levels of CD4 + T and CD16 + CD56 + NK cell were lower compared with control group (23.60% ±6. 42% vs 45.20% ±5.74% ,5.55% ±2. 87% vs 14. 70% ±4. 16% ,respectively ,P ＜ 0.01 ). Conclusion Disorder of cellular and humoral immunity happens in patients with EBV-AHS, which could be implicated in the pathogenesis and progression of EBV-AHS.

ObjectiveTo summarize the early postoperation complications and treatments in children with total anomalous pulmonary venous connection (TAPVC).MethodsThirty TAPVC children who were treated with corrective operation and transferred into PICU were collected.Patients were monitored routinely for electrocardiogram,blood pressure and SpO2.The routine treatment measures included mechanical ventilation,sedation,hemostasis,positive inotropic agents,diuresis,vasodilator,antibiotics and symptomatic treatment.Adrenaline or isoproterenol was used when low cardiac output syndrome appeared and temporary pacemaker was employed in the case of bradycardia.Alprostadil and sildenafil were added instantly after corrective operation when severe preoperative pulmonary hypertension or reactive postoperative pulmonary hypertension was present.ResultsThe early postoperation complication was found in 30 cases of TAPVC,which included 13 cases of pneumonia (43.3％ ),8 of arrhythmia (26.7％ ),7 of low cardiac output syndrome (23.3％),6 of respiratory failure (20.0％ ),4 of pulmonary hypertension ( 13.3％ ),3 of pulmonary edema or atelectasis( 10.0％ ) and 1 of pneumorrhagia ( 3.3％ ).Two cases died postoperatively.The interval of stay in PICU was 1 ～ 21 d and the mean time was (5.95 ± 4.94) d.ConclusionOccurrence of respiratory complications is high among early postoperative complications of TAPVC.Therefore,preventing pulmonary infection and maintaining pulmonary function should be viewed as the key points in early postoperative monitoring and managing.In addition,more attention should also be paid on correction of arrhythmia,prophylaxis and treatment of low cardiac output syndrome and pulmonary hypertension crisis,which may improve the postoperative survival rate and care quality of TAPVC.

In this study, a rapid and sensitive analytical method was developed for the determination of 10 major compounds (procyanidin B1, catechin, procyanidin B2, rutin, isoquercitrin, kaempferol-3-O-rutinoside, astragalin, quercitrin, quercetin, and kaempferol) in Tetrastigma hemsleyanum by using ultra-performance liquid chromatography coupled with triple-quadrupole tandem mass spectrometry (UPLC-MS/MS) in multiple-reaction monitoring (MRM) mode. UPLC-MS/MS assay with negative ion mode was performed on a Waters CORTECS C18 (2.1 mm x 100 mm, 1.6 μm) with the mobile phase consisting of acetonitrile (A) and 0.1% aqueous formic acid (B) in gradient elution at a flow rate of 0.25 mL · min(-1) and the column temperature was set at 45 °C. Under the optimized chromatographic conditions, good separation for 10 target compounds were obtained including chiral isomer procyanidins B1 and B2 were completely separated within 8.5 min. Satisfactory linearity was achieved with wide linear range and fine determination coefficient (r > 0.996 6), the overall recoveries were ranged from 95.44%-110.40% with the RSD ranging from 2.37%-8.69%. It is the first report about simultaneous analysis of 10 major flavonoids components in Tetrastigma hemsleyanum by using UPLC-MS/MS method, which affords highly sensitive, specific, speedy and efficient method for quality control of Tetrastigma hemsleyanum

Objective To compare two effective preparation methods for paclitaxel-loaded lipid microbubbles, and to evaluate the physiochemical properties as for acoustic activated drug delivery. Methods Paclitaxel-loaded lipid microbubbles were prepared with two methods: one was mixed with phospholipids directly (Ⅰ); the other was added in triacetin, then mixed with phospholipids (Ⅱ). Concent ration, size, pH, drug entrapment efficiency, drug-loading amounts of these two kinds of paclitaxel-loaded lipid microbubbles were studied, while drug release with ultrasound and tumor imaging enhanced on rabbit breast tumor were observed. Results There was no significant difference in tumor imaging between two kinds of microbubbles which could be ruptured by low energy ultrasound. Compared with Ⅰ, the mean diameter of Ⅱ decreased significantly ([ 1.07±0.38] μm vs [2.79± 0.41] μm, P<0.01), the surface potencial was higher ([19.10±0.32] mV vs [-5.90±0.21] mV, P<0.01), whereas entrapment efficiency and drug-loading amounts increased markedly ([ 95.00±1.22]% vs [36.10±4.74]%, P<0.01; [5.60±0.11]% vs [0.50±0.04]%, P<0.01). Conclusion The Ⅱ paclitaxel-loaded lipid microbubbles added triacetin have an important clinical value.

Dexmedetomidine(DEX) is a new type of highly selective α₂ adrenergic receptor agonist with multiple effects, such as sedation, analgesic, anti-anxiety and inhibition of sympathetic nervous system activity.DEX is usually used as an anesthetic adjuvant and as an sedative and analgesic in PICU, also possessing effects of preventing and treating emergence agitation, counteracting postoperative shivering and organ protection.This paper summarized the clinical application of DEX in pediatric field.

Pulmonary diseases include infection,injury,asthma,interstitial disease,tumor,hypertension and so on.The mechanisms are not fully understood.Calcitonin gene-related peptide(CGRP),a 37-amino acid neuropeptide,identified in multiple species,has widespread distribution and expression.Although there are no cases that CGRP works in pulmonary disease,many studies have showed that CGRP play an important role on cell line in vitro or on animal models.This brief review provides a preliminary understanding of the diverse biological effects of CGRP and its antagonist in respiratory system.

High altitude pulmonary edema(HAPE) occurs usually in plateau of low oxygen environ-ment,which is a non-cardiogenic pulmonary edema characterized by hypoxic pulmonary hypertension. Children HAPE has an acute onset and rapid progression.It always happens during the first 1 to 3 days ente-ring the plateau and has clear trigger factors such as upper respiratory tract infection,many physical labour and coldness. The pathogenesis is related with hypoxic pulmonary artery contraction,pulmonary epithelial dysfunction,inflammatory response,water transport imbalance in pulmonary epithelium,and genetic polymor-phism.The early symptoms include crying,breath holding,and dry cough. With the disease progressing, patients will present shortness of breath and expectoration of pink foam sputum,and even be unconscious, which results from cerebral edema and is mortal.The treatments of HAPE include bed rest,oxygen therapy, reduction of pulmonary arterial hypertension,hormone,diuresis,prevention of infection and so on.

Objective To explore the effect of calcitonin gene-related peptide (CGRP) on expressions of aquaporin (AQP)1 and AQP 5 in young rats with acute lung injury (ALI) caused by lipopolysaccharide (LPS).Methods Eighty-four young rats were randomly divided into control group,ALI model group and CGRP group.The rats in ALI model group were given intraperitoneal injection of LPS (5 mg/kg)for 2,6,12,24 hours;while the rats in CGRP group were given intraperitoneal CGRP (1 mg/kg) after 1 h injection of LPS.At 2 h,6 h,12 h and 24 h,all rats were sacrificed and lung tissues were obtained.The histopathological changes in lung tissues were evaluated by adopting hematoxylin-eosin staining,and wet/dry(W/D) was measured.The mRNA and protein levels of AQP1 and AQP5 in lung tissues were detected by adopting fluorogenic quantitative PCR (qPCR) and Western blot.Results Pathological stain showed that rats in control group had a normal lung tissue structure,and LPS made lung tissue edema,narrowing the alveolar cavity and inflammatory cell infiltration.CGRP attenuated the effect of LPS on rat's lung.The W/D ratio of lung tissue was significantly higher than that in the control group,and CGRP reduced the W/D ratio of lung tissue.qPCR showed that the mRNA levels of AQP1 and AQP5 from rats in ALI group (0.009 ±0.001 and 0.055 ±0.006)decreased compared with those in the control group (0.035±0.002 and 0.167 ±0.006) and CGRP group (0.024 ± 0.002 and 0.134 ± 0.012) (all P ＜ 0.001).Western blot results showed after 24 h injection of LPS,both AQP1 and AQP5 levels from ALI group (0.397 ± 0.041 and 0.215 ± 0.029) were significantly lower than those in the control group (0.850 ± 0.020 and 0.741 ± 0.032) (all P ＜ 0.001),and their levels in CGRP group (0.593-± 0.065 and 0.461 ± 0.039) were also lower than those in the control group,but higher than those in ALI group (all P ＜ 0.001).Conclusion CGRP can enhance AQP1 and AQP5 levels and reduce pulmonary edema,and it has a protective effect on rats with acute lung injury.

Refractory mycoplasma pneumoniae pneumonia is clinically increasing and has become a common disease that threatens children's lives. This article will introduce the concepts,mechanism,Mycoplasma pneumoniae re-sistance,systemic manifestation,early diagnosis and treatment of refractory mycoplasma pneumoniae pneumonia. Addi-tionally,the reports of pleural effusion and embolism caused by Mycoplasma pneumoniae are increased recently;the achievements on the topic are also reviewed.