Objective: To establish a method for determining the contents of emodin, polydatin, and berberine hydrochloride in Hubai Burrn Tincture. Methods: The samples were separated on an Eclipse XDB-C18 (150 mm × 4.6 mm, 5 μm) column by a gradient elution using acetonitrile and 0.1% phosphoric acid aqueous solution as mobile phase at a flow rate of 1 mL/min. The eluate was detected by double wavelengths: 265 nm for berberine hydrochloride and 306 nm for emodin and polydatin. Column teperature was set at 30 ℃. Results: The linear ranges of emodin, polydatin, and berberine hydrochloride were 6.3-200.0, 9.6-308.0, and 5.0-160.0 ng. The average recovery rates of emodin, polydatin, and berberine hydrochloride were 99.91% (RSD = 1.12%), 99.51 % (RSD = 0.84%), and 99.16% (RSD = 1.17%). Conclusion: This method is accurate, specific, and reliable for the quality control of Hubai Burrn Tincture.

Objective　To approach the relationship betwe en glycosaminoglycan metabolism in cartilages and pathogenesis of KBD.Methods　Rhesus monkey was fed with grains and water from KBD endemic area for 18 months to produce the animal model with KBD . The glyc osaminoglycans in the monkey cartilage were extracted by the improved Dish method of Bitter. Purified glycosaminoglycans were digested with chondroitinaseABC, and the enzymatic digests were analyzed by HPLC. Results　Comparing with those of the control, the glycos aminoglycans in the head of femur, tibia plateau and costal cartilage from the Rhesus monkey fed with grains and water from KBD endemic area were undersulfated . Decreased unsaturated 4-sulphated disaccharide (△Di-4S) from the glycosa minoglycans in the head of femur and tibia plateau and decreased unsaturated 6- sulphated disaccharide (△Di-6S) from the glycosaminoglycans in the costal cart ilage were discovered.Conclusions　Detrimental factors in grains and water from KBD endemic area cause undersulfate of the cartilages glycosaminoglycans from Rhesus monkey. The glycosaminoglycans changes have a direct bearing on the patho logical alterations in morphology on the cartilages from the animal model with K BD.

Objective To investigate the expression of Musashi-2 and CD133 in colonic adenocarcinoma,and their correlation with the occurrence and development of colonic adenocarcinoma thereof. Methods The expressions of Musashi-2 and CD133 protein were detected by immunohistochemical method in 40 colonic adenocarcinoma samples and 15 normal colonic structure samples. The different histological types, TNM staging, lymph node metastasis, serous infiltration, distant metastasis and expressions of Musashi-2 and CD133 proteins in colonic adenocarcinoma tissues were analyzed. Results The positive expression rates of Musashi-2 and CD133 protein were 60%and 27.5%in 40 colonic adenocarcinoma samples and 26.7%and 0 in 15 normal colonic samples, which showed the significantly higher expression rates in colonic adenocarci-noma group than those of control group (χ2=4.850 and 5.156,P<0.05). There were significant differences in expressions of Musashi-2 proteins between different histological stages and TNM staging (P<0.05). The positive expression rate of Musa-hi-2 protein increased as the degree of differentiation decreased and TNM stage increased. There were significant differenc-es in expressions of CD133 proteins between different histological stages and lymph node metastasis (P<0.05). The positive expression rate of CD133 protein increased as the degree of differentiation decreased and lymph node metastasis occurred. Conclusion The expression of Musashi-2 and CD133 may be related with the initiation and development of colonic can-cer, which can be used as the stem cell markers of colonic adenocarcinoma for the further study.

Objective To investigate the clinical characteristics,distribution and drug sensitivity of pathogens causing intravenous catheter-related bloodstream infections (CRBSIs) in pediatric intensive care unit (PICU) so as to use antibiotics reasonably.Methods All patients with CRBSIs in PICU of Guangdong General Hospital from September 2009 to September 2014 were investigated and the drug resistance profiles of pathogens causing CRBSIs were also analyzed retrospectively.Results Between 2009 and 2014,there were totally 10 834 catheter days and 23 episodes of CRBSIs with an incidence of 2.1 infections per 1 000 catheter days.Catheter indwell time ＜ 7 days in 9 cases (39.1％),8 to 14 days in 10 cases (43.5％),14 to 21 days in 4 cases (17.4％).There were 13 strains (56.6％) of gram-positive bacteria,5 strains (21.7％) of gram-negative bacteria and 5 strains (21.7％) of fungi.The main pathogens causing CRBSIs were coagulase negative Staphylococci (7 strains,30.4％),Staphylococcus aureus (3 strains,13.0％),Candida albicans(3 strains,13.0％),Candida parapsilosis(2 strains,8.7％),and Enterobacter cloacae (2 strains,8.7 ％).The susceptibility to Vancomycin,Linezolid and Teicoplanin of coagulase negative Staphylococ cus such as S.epidermidis and to Imipenem,Piperacillin/Tazobactam,Cefoperazone/ Sulbactam and Amikacin of gram-positive bacteria arrived at 100.0％,respectively.The candida were 100％ susceptible to Amphotericin B,5-Flucytosine,Fluconazole and Voriconazole.Twenty-one cases (91.3％) received antibiotic treatment versus no antibiotic in 2 cases (8.7％).The average number of antibiotic kinds administered on the patients with fungal infection was 4.4,bacteria were 1.4.Ten cases (43.5％) treatment with 1 kind of antibiotic,4 cases (17.4％) with 2,4 cases (17.4％) with 3,5 cases (21.7％) with more than 3.Twenty-two cases (95.7％) cured and 1 case died (4.3％).Conclusions The major species of pathogen causing CRBSIs was coagulase negative staphylococci in PICU.It is critical for clinicians to guard against fungal infection because of prolonged catheter indwelling time and more antibiotics administered before indwelling catheter.It is effective way to prevent the CRBSIs by reasonably using antibiotics and shortening the time of catheter indwelling.Monitoring CRBSIs pathogenic bacteria distribution and drug susceptibility helps reasonable administration of antibiotics in the earlier time.

Objective To explore the effect of leg massage during operation on deep venous thrombosis(DVT)in patients of vascular surgery department.Methods Ninety-four DVT patients were randomized into experiment group and control group according to the computer-generated numbers ,46 cases in the latter and 48 in the former.The routine care measures were applied in the control group, while intraoperative leg massage was used in the observation group.The two groups were compared in terms of curative effect and the incidence of DVT.Result The incidences of DVT in the experiment group and the control one were 2.08%(1/48)and 15.22%(7/46)respectively, showing significant difference between them(P<0.05).Conclusion The intraoperative lag massage are significant in the reduction of DVT incidences after operation.

Objective To explore the clinical features,diagnosi s and prognosis of incontinentia pigmenti.Methods Analyzing and summarizing the clinical characteristic, diagnosis and prognosis of neonatal incontinentia pigmenti in 6 neonatal infants that were hospita- lized in our department during the period from January 1 998 to December 2003 were studied,and some relevant literature were reviewed. Results 1.Three of 6 infants were male which was unusual;2.Four infants had typical skin lesions at birth and 1 case at 6 days old.Four cases had typical 3 stages o f skin lesions including the erythematous and vesicular inflammatory stage,verr ucous lesions and hyperkeratosis stage,macular hyperpigmentation stage,but the re was overlap;3.Four infants were complicated by central nervous system involv ement (two cases presented mental retardation,2 infants were temporary damage). Two cases were complicated by ocular manifestations ( one case had optical nerve atrophy and blind in left eye,the other had severe bilateral retinal lesions); 4.On specific examination 5 infants were diagnosed by skin biopsy.Gene analysis was made in 1 case,but we didn′t find the mutations of NEMO. Conclusions Incontinentia pigmenti is a rare X-linked dominant multisystem disease.It may be misdiagnosed in the initial stages.Except typical clinical features,skin biops y and gene analysis are main evidence for diagnosis.Early detection and interven tion are important for prognosis. J Appl Clin Pediatr,2005,20(2):123-125

Objective To improve the recognition of intramedullary spinal abscess by a case of congenital dermal sinus with intramedullary spinal abscess and reduco the incidence of congenital dermal sinus with intramedullary spinal abscess.Methods Clinical,laboratory data and image of a confirmed case about one infant of congenital dermal sinus with multiple intramedullary spinal abscess were investigated,the related literature was reviewed.Results In this case,when the infant with congential dermal sinus had infection,he failed to gain antibiotic therapy, timely surgical treatment,his infection had diffused, and multiple intramedullary spinal abscess flared up.Conclusions Intramedullary spinal abscess is a rare disease.If treatment is delayed, the prognosis is poor and the mortality rate is high.MRI is the ideal investigation for diagnosis.Intramedullary spinal abscess can happen subsequent to congenital dermal sinus with infection, and cause neurological sequela. So an infant with congenital dermal sinus should be offered to avoid complication caused by infection.

Background Experimental autoimmune uveoretinitis(EAU)is proved to be an organ-specific,T lymphocyte-mediated autoimmune and self-limited disease.Research showed that CD4+CD25+T cell may play important regulation on the course of events,but its mechanism is pending for further study. Objective Present study was to investigate the potential role of CD4+CD25+T cell in the pathogenesis of EAU. Methods Retinal Santigen(S-Ag)was isolated from bovine retinas according tO the procedure as Caspi's previously describe.0.1 ml Santigen(50μg)emulsified with complete Freund'S adjuvant was injected on footpad of 24 inbred adult female Lewis rats aged six to eight-week-old to induce EAU,and 4 normal Lewis rats were as normal control group.Slim-lamp examination was performed to observe the ocular manifestation.Retinal section was prepared in 7,12,15,21 days aher injection for the pathological examination.The pathological grading was on the lnoki'S method.The retinas,inguinal nodes and spleens of rats were obtained in 7,12,15,21 days after injection and the cellular suspension was prepared.Expression of CD4+CD25+T cells on cellular suspension was assayed using flow eytometry.This study complied with the Standard of Association for Research in Vision and Ophthalmology. ResuRs The obvious inflammatory response of the anterior segment was found in S-Ag injected eyes from 7 days through 21 days.The most serious inflammation was found in 12-15 days under the slim-lamp.The hemotoxylin and eosin staining showed the higher pathological grading from 12 to 15 days after injection,showing significant difference in comparison with 7 days and 14 days(P=0.000).In EAU model rats,expressions of CD4+CD25+T cells was significantly increased in retinas, inguinal nodes and spleens in 15 days after injection,showing evidently differences in comparison with control rats(P=0.000). Conclusion The expression level of CD4+CD25+T cells in inflammatory tissue is associated with the inflammation procedure in EAU model rats.This study indicates that CD4+CD25+T cells may play a role in the development of EAU.

Objective To provide immunological evidence for clinical transfer of chemical extracted acellular nerve allografL Methods One hundred and twenty-eight BALB/C mice were randomly divided into 4 groups of equal size according to their different treatments:negative contrast group(NC),fresh autograft group(AG),fresh allogeneic nerve group(FN)and chemical extracted aceflular allogeneic nerve group(CEN).Then we implanted various kinds of nerve grafts into the thigh muscle of BALB/C mice in corresponding groups.At 3,7,14,28 days postoperatively,8 mice from each group were killed each time to harvest their spleens,from which T lymphocytes were collected.Theu monoclonal antibodies(CD3,CD4 CD8 CD25,IL-2,IFN-γ, TNF-α)were added into the suspension.Then fluorescence.activated cell sorting(FACS)was used to determine the positive rates of cells combined with the above monoclonal antibodies. Results There were no statistically significant differences between CEN group,NC group,and AG group,but indexes of FN group were significantly higher than those of the other 3 groups at corresponding time points. Conclusion There is no obvious immune reiection of chemical extracted acellular nerve allograft when compared with fresh nerve autograft.

AIM:ToinvestigatetheexpressionofFoxp3+regulatoryTcells(Foxp3+Tregs)andprogrammed death receptor 1 (PD1) in gastric cancer tissues and their association with clinicopathological factors and prognosis of the patients.The correlation between the 2 molecules was also analyzed at the same time.METHODS: The tumor sections from 111 gastric cancer patients were stained for Foxp3 and PD1 by the method of immunohistochemistry.The associations of the expression levels of these 2 molecules with clinicopathological factors involved in the disease progression and progno-sis were statistically analyzed .The relationship of their expression was detected.RESULTS:Foxp3 +Tregs and PD1 were expressed in the gastric cancer tissues, and PD1 was expressed in the tumor infiltrating lymphocytes ( TILs) .The expres-sion of Foxp3 and PD1 was correlated with lymph node metastasis, clinicopathological stage and prognosis of gastric cancer patients.The expression of these 2 determinants in the patients with lymph node metastasis and an advanced clinicopatho-logical stage was distinctly higher ( P<0.05 ) .The patients with positive expression of the 2 indexes presented a lower overall survival rate and worse prognosis (P<0.05).A significantly positive correlation between the infiltration of Foxp3 +Tregs and the expression of PD1+TILs was also observed (P<0.01).CONCLUSION:Foxp3 +Tregs and PD1 +TILs co-infiltrate in the gastric cancer tissues, which can be used as biological markers to predict the disease progression and prog-nosis.