ObjectiveTo observe the changes of amplitude-integrated electroencephalogram (aEEG)of neonatal asphyxia with different Apgar scores,and to investigate the diagnostic value of aEEG for hypoxicischemic encephalopathy(HIE) in neonatal asphyxia.MethodsaEEG monitoring were detected on 56 fullterm asphyxia neonates who were hospitalized in our neonatal department from Dec 2010 to Oct 2011.According to 1 minute Apgar score after birth,56 cases were divided into two groups:observation group in which 36 cases with Apgar score 0～7,and control group in which 20 cases with Apgar score 8～10.aEEG monitoring was done within 6 h,2 d,3 d,7 d of each neonates after birth,and the changes of aEEG were analyzed and the diagnostic value on HIE were evaluated.ResultsAmong 20 cases in the control group,the aEEG results in 6 hours after birth were 17 cases (85％) had normal aEEG results,3 cases( 15％ ) mildly abnormal,nd no one severely abnormal.The aEEG results of patients in observation group(36 cases) were 18 cases(50％ ) had normal aEEG results,13 cases(36.1％ ) mildly abnormal,and 5 cases( 13.9％ ) severely abnormal.The abnormal rate in observation group was significantly higher than that of the control group ( x2=5.3,P＜0.001 ).There were 34 HIE patients in the total 36 cases of observation group,whose aEEG monitoring results in 6 hours after birth were associated with HIE clinical grading( Spearman's rank correlation coefficient was 0.867,P＜0.01 ).Dynamic aEEG monitoring for 56 patients showed that 21 cases had abnormal aEEG in 6 hours after birth,in whom 15 cases(71.4％ ) could returned to normal after 48～72 hours after birth,and there were only 4 case (7.1％ ) still had severely abnormal aEEG results in the seventh day after birth.ConclusionThe aEEG dynamic monitoring for full-term HIE neonates after birth enhances early prediction of HIE.

Objective To ivestigate the clinical feature and therapeutic ways of the gastroduodenal ulcer perforation(GDUP) in aged patients.Methods The clinical data of 58 GDUP patients with age more than 60 gears treated in recently tweenty yesas in our hospital were analysed retrospectively. Results Among the 58 patients,56 patients underwent operation,46cured and 12 died.Of the 12 patients,dead of cardiorespiratory function failture in 5 ; septic shock in 4 ;and renal failturl in 3 .Conclusions Aged patients with GDUP would be operated as early as possible.Repair of the performed ulcer plus modified vagotomy is a better choice.

Objective To investigate the therapeutic efficacy of endoscopic sphincterotomy (EST) for treatment of choledocholithiasis. Methods The clinical data of 326 patients receiving EST from July 1990 to January 2009 were retrospectively analyzed. Long-term follow-up was performed in 195 of the 326 patients. Results The common bile duct stones were removed in 301 cases (92. 33%).The occurring rate of complications after EST was 6.13% (20/326). Of the 20 patients, 10 had pancreatitis (3.07%), 7 hemorrhage (2. 14%), 4 cholangitis (1. 220%) and 1 duodenal perforation (0.09%). Only 1 patient died. The follow-up showed that 20 patients had reflux cholangitis and 18 recurrence of common bile duct stones. Conclusion Therapeutic endoscopy is safe and effective for treatment of choledocholithiasis.

Objective To establish a rapid,accurate and specific method to detect the common mycobacteria based on multiplex real-time PCR.Methods The dual priming oligonucleotide ( DPO)primers and TaqMan probes labeled with FAM,ROX,HEX or JOE fluoresceins at 5' end and eclipse at 3' end respectively were designed to detect the 16S rRNA of mycobacteria.Both specificity and sensitivity were estimated on multiplex real-time PCR detecting genome DNA from 4 mycobacterial model species.Sixty eight early morning sputum specimens collected from hospitalized patients in the Red Cross Hospital of Hangzhou were detected by multiplex real-time PCR,bacterial culture and smear microscopy simultaneously.The positive rates were analyzed by chi-square.Results Mycobacteria including Mycobacterium tuberculosis and three common non-tuberculosis mycobacteria spp.were identified by multiplex real-time PCR accurately and specifically,with the limited load at 101 cfu/ml.In 68 sputum specimens,31 were positive (positive rate 45.6％ ) by this method,which was significant higher than that by smear microscopy ( positive rate 14.7％,x2 =15.4,P ＜0.05 ).The positive cases were identified as 28 Mycobacterium tuberculosis,1 Mycobacterium avium and 2 Mycobacterium intracellulare in agreement with the culture results.One case,which is detected by culture,but not by PCR,was identified as Mycobacterium chelonae by sequencing.Conclusion The multiplex real-time PCR characterizing as sensitive,specific and time-saving for Mycobacterium tuberculosis and common non-tuberculosis mycobacteria could be chosen as the rapid laboratory test of mycobacterial infection.

Objective To summarize the experience of operative treatment of all types of congenital choledochal cysts (CCC).Methods Clinical data from 52 operated patients with CCC were analyzed retrospectively.(Operations) included excision of the cyst with Roux-en-Y hepaticojejunostomy in 38 cases,partial excision of the cyst with choledochojejunostpmy with Roux-en-Y loop in 2 cases etc. Results 52 cases were cured and left hospital . Among them, 45 were followed up and 42 cases had a good recovery. Conclusions (Resection) of the cyst with Roux-en-Y hepaticojejunostomy is recommended as the therapy of choice in selective operation.Adherence to the indications,the method of excision and the principle of Roux-en-Y (hepaticojejunostomy) are the key factors to increase the effect of the operation.

OBJECTIVE To develop a real-time fluorescence PCR assay to detect the genes encoding thermolabile(hemolysin)(TLH),thermostable direct hemolysin(TDH) and TDH-related hemolysin(TRH) of Vibrio(parahaemolyticus).METHODS The genes of TDH and TRH were selected as target ones of thermostable direct and TDH-related hemolysin,and TLH gene as a specific genomic marker for V.parahaemolyticus.Designed and synthesized the primers and Taqman probes,we investigated 487 stool samples of doubt foodborne illness patients by real-time fluorescence PCR.RESULTS The sensitivity of the assay for TLH and TDH was 1.0?10~2copies,but the sensitivity of TRH was 1.0?10~3copies. Among 487 samples,112 V.parahaemolyticus strains were found;101 samples of these strains showed the production of TDH;none of them was positive for TRH.CONCLUSIONS The Taqman PCR is a rapid and sensitive method to detect the TLH,TDH and TRH of V.parahaemolyticus,it is well suited for screening large numbers of samples at the same time;and TDH is one of the primary virulence factors in clinical isolated V. parahaemolyticus.

To develop a low-cost, reliable, easy-to-maintain and practical instrument for protein purification. Some ultraviolet luminescent diode was used to provide 280 nm light source, and high-sensitivity S1336 photo-electric detector was employed for real-time monitoring of purified protein solution flowing through quartz cell to supervise the concentration of the protein. The instrument gave voice alarm and stopped working in case the protein concentration was less than the standard one. The lower SCM monitored the liquid level of the protein collecting cup and the position of loading arm through laser infrared distance sensor, so that a cup full of protein might be replaced by another empty cup. The instrument involved in Samsung S5PV210 embedded master computer, Wince6.0 operating system, Keil4.0 and VS2005. Trials proved that the instrument could perform real-time monitoring and curve display of dual-channel ultraviolet absorption, and could realize auto collection of 735 ml protein solution and up to 5-hour standby. The instrument developed has simple structure, high reliability and easy maintenance, and meets the desired require-ments.

Objective To investigate the incidence,distribution patterns,and influencing factors of cerebral microbleed (CMB) in Chinese adult patients with moyamoya disease.Methods Thirty consecutive patients with moyamoya disease confirmed by digital subtraction angiography from the Nanking Stroke Registry Program were included.All patients performed conventional MRI sequences (3.0 T) and susceptibility-weighted imaging.The clinical data,such as medical history,systolic blood pressure,diastolic blood pressure,mean arterial pressure,and white matter lesions were collected.Their fasting blood glucose levels and fibrinogen levels were detected.The numbers of CMB lesion,distribution information,and their relationship with various clinical parameters in patients with moyamoya disease were analyzed.Results Among the 30 subjects included,14 CMBs were detected in 10 patients from 11 hemispheres.The distribution of the lesions was mainly in deep brains (71.4%),especially in the periventricular white matter (50.0%).There was no statistical difference in age,gender,hemorrhage symptoms,blood pressure,white mater lesions,and plasma fibrinogen levels between the CMB positive group and CMB negative group.However,the fasting blood glucose levels in the former was significantly higher than those in the latter (8.0 ± 4.1 mmol/Lvs.4.8 ± 0.4 mmol/L;P =0.035).Multivariate regression analysis showed that the increased fasting blood glucose level was an independent risk factor for the occurrence of CMBs in adult patients with moyamoya disease (OR = 10.992,95% CI 1.325-91.218;P=0.026).Conclusions The CMB lesions are susceptible to Chinese adult patients with moyamoya disease in deep brains,especially in the periventricular white matter.The fasting blood glucose level may influence the incidence of CMBs in patients with moyamoya disease.

Objective To analyze the genetic characterization of VP1 region of Coxsackie virus A10(CVA10)isolated from clinical specimens of hand, foot and mouth disease(HFMD) patients in Shandong Province. Methods Clinical specimens were collected from some of HFMD patients from 2008 to 2009. The virus was isolated by cell culture. Total RNA was extracted, and the VP1 genes of the isolates were amplified by reverse transcription-polymerase chain reaction (RT-PCR) and sequenced. The genotypes were identified by molecular typing method and bioinformatics analysis.Homologous comparison and phylogenetic analysis of representative CVA10 strains were performed.Homologous comparison between the Shandong isolates and strains obtained from GenBank were performed and phylogenetic analysis of some representative CVA10 strains were performed. Results Three hundred and thirty viruses strains were isolated from 760 clinical specimens collected from HFMD patients, and 17 of them were identified as CVA10. The homologies of nucleotide and amino acid of the 17 CVA10 strains were 82.3%-100.0% and 94.2%-100.0%, respectively. Compared with the prototype strain of CVA10 (Kowalik/USA/2003), the homologies of nucleotide and amino acid were 75.6%-76.8% and 90.2%-93.2%, respectively. Interestingly, Shandong CVA10 strains were clustered into two distinct subgroups in the phylogenetic tree. Conclusions CVA10 is one of the causative agents of HFMD. Two independently circulating subgroups of CVA10 exist in Shandong province.

Objective To investigate the variation characteristics of adenovirus type 55 (HAdV-B55) gene on plateaus.Methods Throat swabs were collected from HAdV-B55 infected patients and used for virus isolation in HEp-2 cells.The whole-genome sequence was obtained by PCR and sequencing.HAdV-B55 gene sequence was blast with the previously reported virus.Results HAdV-B55 strains were isolated from throat swabs, which were named LS89/Tibet/2016.The whole-genome sequence was obtained and submitted to GenBank with the accession number of KY002683.No large fragment gene recombination was found between this HAdV-B55 strain and previous strains, and the sequence similarity with QS-DLL strain was 99.9%.Conclusion This study provides more information for the evolution patterns of adenovirus 55 and will contribute to the prevention and control of HAdV-B55 infection in the future.