ObjectiveTo explore the relationship between the changes of the blood platelet counts (BPC) in children with cerebral palsy (CP) and pather mechanism of CP in traditional Chinese medicine.MethodsBPCs of CP children and healthy children were analyzed with statistical method.ResultsThe BPC of CP children was higher than that of the healthy children ( P<0.05).ConclusionThe BPC of CP children is higher than that of the healthy children, and this is consistent with the theory of traditional Chinese medicine about pathogenic mechanism of CP that CP is related to blood stasis.

Objective To study the effect of morin on LPS induced acute lung injury mouse model and its mechanism .Meth‐ods Thirty male C57B/L mice were randomly divided into control group ,LPS group and LPS+ morin group ,with 10 in each group .5 mg/kg LPS was instilled into the lung from an trachea intubation in LPS group and LPS+morin group .Then the mice in LPS+morin group received an intraperitoneal injection of morin (40 mg/kg) every day for the next 3 d .Others received an equal a‐mount of saline .After 72 h ,the mice were sacrificed .The bronchoalveolar lavage fluid (BALF) was collected and centrifuged;the sediments were stained with Wright‐Giemsa for total cell and neutrophil count and the supernates were prepared for ELISA .The wet and dry weight of lung was weighed to calculate the wet/dry weight ratio .HE staining was performed to examine the pathologi‐cal change of lung .Western blot was used to determined the expression of TLR4 ,IKK and NF‐κB .Results Intratracheal instillation of LPS successfully established ALI model in mouse .LPS caused significant pathological changes including inflammatory cells infil‐tration ,alveolar septa thickness ,hemorrhage and edema .The wet/dry weight ratio ,the total cell count ,neutrophil count ,TNF and IL‐1βlevel in BALF ,and the expression of TLR4 ,NF‐κB ,and IKK were all increased significantly (P<0 .05) ,which were allevia‐ted by intraperitoneal injection of morin .Conclusion Morin can dampen the inflammatory response during LPS induced ALI in mouse ,which is potentially attributed to its inhibitory effect on the activation of NF‐κB .

Objective To increase the preoperative diagnosis rate,reform the operation and improve the prognosis in children with ganglio -nauroma.Methods Clinical data of 20 children with ganglioneuroma hospitalized in Tongji hospital of Huazhong university of science and technology from Nov.1986 to May.2006 were reviewed(male 9,female 11).The biological,clinical and pathological characteristics of patients were analyzed.Results Among these 20 patients,15 cases were discovered to have no clinical symtom.The B ultrasound showed low-echo in all 16 cases.Under CT scan,20 cases showed low to moderate density shadows,and the distinct enhancement in these tumors was noted with enhanced CT scan.Fourteen cases undergoing MRI all showed low signals in T1W1 and inhomogeneous high signals in T2W1.All patients underwent surgical operations,completely removed in 14 cases,partly in 4 cases, and biopsy was performed only in 2 cases.Fifteen cases were followed up from 7 months to 6 years,tumor recurrence happened only in 1 case.Conclusions Ganglioneuroma is mostly found in chest-abdominal sympathetic chain,it grows up slowly and seems to be innocent.The imaging data of B ultrasound,CT and MRI can give helpful information in the diagnosis of ganglioneuroma.We shall pay attention to the choice of incision and avoidance of injuring the important blood vessels during the operation.Children who have neuroblastoma cells should receive prophylactic chemoprophylaxis treatment and the result of long-time follow up is satisfactory.

Objective:To measure the effective optical zone (EOZ) after small incision lenticule extraction (SMILE) by using corneal topographic map and modulation transfer function (MTF), and to analyze the related factors affecting the EOZ.Methods:A retrospective observational case series study was performed.Sixty-two myopic patients (62 eyes) who underwent SMILE between December 2015 and July 2017 in Jinan Mingshui Eye Hospital were enrolled.The EOZ was measured by using tangential corneal curvature topographic map and MTF pre- and 6-month post-operatively.The repeatability of data was determined by intraclass correlation coefficient (ICC) and Cronbach Alpha coefficients; the agreement of data was identified by using Bland-Altman plot; the correlations between EOZ and surgical parameters were analyzed by utilizing Pearson correlation coefficient.This study protocol was approved by the Ethics Committee of Jinan Mingshui Eye Hospital(No.2015[013]). Written informed consent was obtained from each patient before surgery.Results:The ICC and Cronbach Alpha coefficients of EOZ measured by corneal topographic map were greater than 0.9, which showed high intraobserver repeatability.The Bland-Altman plots displayed relatively good agreement between the two methods.The 95% limits of agreement was -0.49 to 0.89 μm.Six months after SMILE, the EOZ measured by tangential corneal curvature gradient topographic map was (5.32±0.25)mm, which was (1.18±0.25)mm smaller than predicted optical zone, and the EOZ measured by MTF method was (5.07±0.32)mm, which was (0.20±0.35)mm smaller than corneal tomographic EOZ, and the difference was significant ( t=-4.487, P<0.01). A negative correlation was found between the EOZ and attempted refractive correction ( r=-0.364, P=0.004). The positive correlations were found between the EOZ and ΔKm or ΔQ 6 months after SMILE ( r=0.367, 0.514; both at P<0.01). Conclusions:The EOZ measured by tangential corneal curvature topographic map after SMILE is of high repeatability and is consistent with the result calculated by MTF method.The EOZ is significantly reduced after SMILE.

AIM:To investigate the role of minocycline on glutamate uptake in the periventricular zone and its putative mechanism after hypoxic exposure in neonatal rats.METHODS: A model of hypoxic-ischemic brain damage (HIBD) was developed by putting postnatal 1 d rat pups in 5%O2 for 3.5 h.The glutamate level in periventricular zone was measured by liquid chromatography coupled with tandem mass spectrometry assay ( LC-MS/MS) after hypoxic exposure for 4 h and 1 d.The dynamic changes of glutamate transporters EAAT1, and EAAT2 during developmental period in periventricular zone were determined by Western blot.Moreover, the expression of EAAT1, EAAT2, Iba-1, IL-1β, TNF-αand TGF-β1 was also detected by Western blot after hypoxic exposure for 4 h and 1 d in that region.The effects of mino-cycline on all parameters mentioned above were tested after minocycline treatment at the same time points and in the same region.RESULTS:After hypoxic exposure, glutamate level was increased, but it was decreased after minocycline treat-ment.EAAT1 and EAAT2 kept a low expression level at the first postnatal week, but a predominant elevation was found at the end of the second postnatal week.The expression of EAAT1, EAAT2, Iba-1, IL-1βand TGF-β1 was increased at 1 d after hypoxic exposure.EAAT1 and TNF-αexpression was significantly up-regulated, while EAAT2 was down-regulated af-ter minocycline treatment.CONCLUSION: Minocycline inhibits the increase in the glutamate level after hypoxia in periventricular region of the neonatal rats.The mechanism may relate to the selective regulation of glutamate transporters, rather than the inhibition of neuroinflammation in periventricular zone.

ObjectiveTo determine the potential natural foci of new bunyavirus,and isolate and identify the new bunyavirus strain in sera from suspected new bunyavirus-infected patients.MethodsImmunofluorescence assay was used to detect the antigens of new bunyavirus in different tissue specimens of wild rodent animals in Tiantai area of Zhejiang province.Fluorescence quantitative real-time RT-PCR was applied to detect the viral nucleic acid in sera of suspected new bunyavirus-infected patients and the amplification products were analyzed by sequencing.The new bunyavirus in the pateints'sera was isolated using Vero cells.Using nucleocapsid protein encoding gene of new bunyavirus as the target gene,the isolated suspected new bunyavirus strain was identified by RT-PCR and sequencing of the amplification product.Moreover,sequence identity of the amplification product of nucleocapsid protein encoding gene of new bunyavirus was analyzed and compared.ResultsOf the 70 wild rodent animals,5.71% were positive in the immunofluorescence assay.The fluorescence quantitative real-time RT-PCR confirmed that two of the four detected patients'serum specimens were positive.One suspected strain of new bunyavirus was isolated from one pf the two positive patients'serum specimens.The results of RT-PCR and sequencing confirmed that the viral strain exactly belongs to new bunyavirus with 92.2% sequence identity to that of the new bunyavirus isolates in Hubei province but distinct with the new bunyavirus isolates from other areas in China.ConclusionThe presence of natural foci of new bunyavirus and new bunyavirus-infected patients in Zhejiang province are firstly confirmed by this study.There is a geographical diversity of the distribution of new bunyavirus in different groups.

Aim To select a potential biomarker for early lung cancer diagnosis and targeted therapy by using the cancer specific bounded peptide ZS-6 which had already been obtained from the laboratory.Methods The peptide ZS-6 marked by biotin was used as a probe to pan out the human lung cancer cDNA phage display library,after 4 rounds of subtraction panning,the specific binding clones of ZS-6 were identified.After amplification and purification,then those DNA sequences were identified and analyzed with bioinformatics.Results 18 phage clones were identified to the specific peptide ZS-6 and the DNA sequence showed one of them was an unknown new gene while the others were known tumor related genes.Conclusion A tumor biomarker selected from human lung cancer cDNA library provides a potential tool for early lung cancer diagnosis and therapy.

Objective To investigate the feasibility of constructing tissue engineering urethral substituted graft using human lingual keratinocytes and natural derived scaffold.Methods From Oct.2009to Jan.2010,ten patients with anterior urethral stricture were enrolled in this study.A 0.5 × 0.8 cm lingual mucosa was harvested during the operation.Lingual keratinocytes were then isolated and cultured from the mucosa.AE1/AE3 antibody was used to identify the lingual keratinocytes.Keratinocytes were collected and seeded onto three types of scaffold including dehydrated BAMG,liquid stored BAMG and 4-layer SIS product,with the density of 1 × 107/ml at passage three.After being cultured for seven days in vitro,H&E staining and Electronic Scan Microscopy were used to evaluate the compound matrixes.Results No complication occurred in the patients after operation.The primary passage of confluence lingual keratinocytes appeared in a typical cobblestone shape after being cultured for 14 days in vitro.The proliferation rate of these cells increased rapidly during the three passages.However,it decreased significantly in the 4th passage.H&E and Electronic Scan Microscopy examinations showed that few cells grew on the surface of the liquid stored BAMG.Nevertheless,multiple keratinocytes layers could be seen in dehydrated BAMG and 4-layer SIS.Meanwhile,cellular infiltration could be observed in SIS sections.Conclusions Human lingual keratinocytes could be the alternative of seeding cells for constructing tissues for engineering the urethra.These cells exhibited good compatibility and are adhesive to the SIS or BAMG.The compound matrix,which used human lingual keratinocytes and natural derived scaffold,may meet the clinical need of urethral disease in the future.

Objective To analyze the positive rate of specific distribution characteristics in Rh blood group antibody,analyze the clinical significance of Rh blood group antibody and rules.Methods The micro column gel anti globulin technique was used to screen and identify irregular red blood cell antibodies,for patients with Rh blood group antibody,monoclonal anti-D,anti-C,anti-c, anti-E,anti-e were used to identify Rh blood group antigen to confirm the accuracy of detection.The antibody titer,Ig-type and 37℃ reactive were used to determine its clinical significance.Through asking pregnancy history,history of blood transfusion,under-standing whether the same specificity of the antibody in maternal plasma if the patient was newborn,the causes of antibody were an-alyzed.Results In 109 000 patients,Rh blood group antibodies were detected in 96 cases,the positive rate was 0.088%,which has a history of pregnancy in 68 cases,5 cases had history of blood transfusion,both pregnancy history and history of blood transfusion in 6 cases,1 7 cases of neonatal maternally derived antibody.Antibody specificity:65 cases of anti-E(67.710%),12 cases of anti-cE (12.500%),8 cases of anti-D (8.330%),7 cases of anti-c(7.291%),2 cases of anti-C (2.083%),2 cases of anti-e(2.083%).96 cases of Rh blood group antibodies were IgG or IgG+IgM class,37 ℃ reaction could be with the corresponding antigen of red blood cell,antibody titer between 4-2 048.Conclusion Anti-D detection rate shows a trend of gradually decreasing.In Rh blood group antibody detection,anti-E and anti-cE account for an absolute majority.Alloimmune caused by pregnancies and blood transfusion is the main reason of Rh blood group antibody production from Rh blood group antibody.Neonatal maternal passive getisa Rh-HDN is the main pathogenic antibody.

Objective To evaluate the immune reconstitution of HIV-1 infected individuals after long-term highly antiretroviral therapy (HAART). Methods Twenty-five HIV-1 infected individuals receiving HAART, 17 without HAART and 15 healthy controls were included in the study. CD4 +T, CD8 +T,CD8/human leukocyte antigen DR (HLADR) + T, CD8/CD38 +T cells, and the expression of CD127 on CD3 +T cells from peripheral blood samples were measured by flow cytometry. IL-7 in peripheral blood was measured by enzyme-linked immunosorbort assay (ELISA) in HAART group. t test was performed to compare the measurement data among the groups. Results Before HAART, the count of CD4 + T cells in HIV-1 infected group was lower than that of the healthy control (t =9. 12, P ＜0. 01 ), while the counts of CD8 + T,CD8/HLADR+T, and CD8/CD38 +T cells were higher than those of the healthy control (t = 4.48, 4.89 and 3.88, P＜0. 01 ). Ater 7 years' antiviral therapy, CD4 +T cells increased, CD8 +T cells decreased, but both of them didn' t reach to the normal levels ( t = 2.66 and 2.43, P ＜ 0.05 ). While the counts of CD8/HLADR+T cells and CD8/CD38 +T cells almost reached to the normal levels (t = 0. 86 and 1.39, P ＞0.05). Before HAART, the concentration of IL-7 in HIV-1 infected group was higher than that in healthy controls (t =5.31, P ＜0.01 ). It decreased with HAART, but was still higher than the normal level (t =2. 81, P ＜ 0. 05 ). The expression of CD127 on CD3 + CD8 + T cells in non-HAART group was significantly lower than that in healthy control ( t =- 6.01, P ＜ 0.01 ), while that in HAART group was higher ( t = 2.32,P ＜0.05), but still not reached to the normal level ( t = 4.49, P ＜ 0. 05 ). CD127 expression on ( CD45RA + ) CD3 + CD8 + T cells almost increased to the normal level ( t= 0. 28, P ＞ 0. 05 ), while that on ( CD45RO + ) CD3 + CD8 + T cells was still remarkably lower than the normal ( t = 4. 86, P ＜ 0. 05 ). Conclusion Long-term HAART can partially restore the count and function of lymphocyte subsets in HIV-1 infected individuals, and the abnormal immune activation can be inhibited.