Objective To confirm the safety and nutritional efficacy of high branched-chain amino acids through a pragmatic study allowing its use as an alternative to 15AA in patients with liver dysfunction. Methods The study was performed as a randomized, prospective trial. Eighty two patients with liver dysfunction undergoing operation were randomly assigned to receive high branchedchain amino acids or 15AA as part of their TPN regimens for 7 days. Daily parenteral intakes of energy nitrogen and lipid were equal in the two groups. Results Efficacy analysis showed that high branched-chain amino acids were as efficient as 15AA in avoiding protein catablosim. No serious adverse event was reported in the two groups. For hematology, renal, hepatic safety criteria and for the vital signs,no significant difference was observed between the 2 groups. No significant difference was observed concerning nitrogen balance and protein catablosim. For peripheral immunoglobulin and lymphocytes, a statistically significant difference was observed between the high branched-chain amino acids and the 15AA groups. Conclusion High branched-chain amino acids is new, safe and efficient amino acids for parenteral nutrition.

Objective The purpose of this study was to assess the value of prenatal ultrasound diagnosis for fetal ear auricle malformations.Methods The coronal and sagittal planes of fetuses ears were obtained prospectively in 6239 singleton fetuses in the First Affiliated Hospital of Shantou University Medical College for the period from 2012 February to 2015 December,the ultrasound images and pregnancy outcomes were analyzed in 11 cases of fetuses ear auricle malformations diagnosed prenatally.Results Eleven Cases of fetuses ear auricle malformations include with 7 cases of microtia,3 cases of low-set ears and 1 case of anotia.Eleven cases were combined with other structural malformations were as followings,3cases with craniocerebral congenital malformation,5 cases with dentofacial deformity,5 cases with malformation of heart,3 cases with limb deformity.Cordocentesis was performed in 7 cases among which 6 with abnormal karyotype,including 2 cases of trisomy 21,2 cases of trisomy 13,2 cases of trisomy 18,1 case of 22ql 1 abnormalities.Compared with the postpartum facial examination,prenatal ultrasound correctly diagnosed 10 cases of fetal ear auricle malformations,missed diagnosis 1 case of microtia.Conlusions Fetus with ear auricle abnormalities have characteristic prenatal ultrasound imaging;prenatal ultrasonography can provide reliable information in the diagnosis of this disease.This study suggests that antenatal ear auricle length measurements might be a promising sonographic screening method for the detection of abnormal karyotype in pregnancy.

Objective To detect Pneumocystis carinii (Pc) DNA by loop-mediated isothermal amplification (LAMP). Methods After injected with hydrocortisone acetate for 8 weeks, the bronchoalveolar lavage fluid (BALF) of Wistar rats were collected and a portion of BALF were examined for identifying Pc organisms using microscope. Then Pc DNA was extracted by phenol-chloroform extraction. Four primers which recognized 6 distinct regions on the mtrRNA gene of Pc were designed and used for LAMP assay. To evaluate the specificity of the assay, M. tuberculosis, M. pneumoniae, C. pneumoniae, P. gondii and rat leucocyte were used as negative controls. To compare the sensitivity of the LAMP to that of conventional PCR, Pc DNA were 10-fold serially diluted and was amplified by LAMP and PCR. LAMP results were judged by naked eye, electrophoretic analysis and restriction digestion. Results Pc organisms were detected from BALF of rats injected with hydrocortisone acetate. After LAMP reaction, positive signal was observef rats injected with hydrocortisone acetate. By contrast, no positive signal was observed for the negative controls in the study. The amplified product digested by restriction enzyme demonstrated 3 bands (82, 135, 189 lip) upon agarose gel electrophoresis, in good agreement with the predicted sizes. The detection limit of LAMP assay was 1 pg/μl of Pc DNA per reaction and that of PCR was 10 pg/μ1 of Pc DNA per reaction. Conclusion LAMP assay has usefulness for rapid detection of Pc.

Objective To localize and characterize the plasmid protein pORF5 in the Chlamydia trachomatis(Ct) infected cells. Methods The open reading frame encoding for pORF5 protein from the Ct plasmid was amplified and cloned into the pGEX-6p vector. The recombinant plasmid pGEX-pORF5 was transformed into XL1-blue E. coli to express fusion protein with the glutathione-s-transferase (GST). After purified with Glutathione Sepharose 4B beads, the pORF5 fusion protein was used to immunize mice to make monoclonal and polyclonal antibody. The antibodies were used to localize the endogenous pORF5 protein and detect the expression pattern in Chlamydia-infected cells using an indirect immunofluorescence assay (IFA). At the same time, ELISA was used to determine whether pORF5 plasmid protein was expressed and immunogenic during Ct infection in humans. Results pORF5 was detected a dominant signal in the cytosol of the Chlamydia-infected cells with a pattern similar to that of anti-CPAF. pORF5 also appeared in the RBs and EBs in small quantity. Athough pattern was similarly, pORF5 did not overlap with CPAF. pORF5 protein was strongly recognized antiserum in an ELISA. Conclusion The pORF5 plasmid protein was identified as a secreted protein with good immunogenicity, pORF5 gene was to express the endogenous target protein during human infection.

Objective To detect Toxoplasma gondii DNA by loop-mediated isothermal amplification(LAMP). Methods DNA was extracted by phenol-chloroform extraction from T. gondii tachyzoites. Four primers which recognized 6 distinct regions on the B1 gene of T. gondii were designed and used for LAMP assay. To evaluate the specificity of the method, Plasmodium vivax, P. falciparum, Pneumocystis carinii, Schistosoma japonicum, and mouse leucocytes were used as controls. The parasite extract (T. gondii) was 10-fold serially diluted for evaluating the sensitivity of LAMP, and was amplified by LAMP. LAMP results were read with naked eye and analyzed by electrophoresis. Results After LAMP reaction, positive amplification was observed with T. gondii, but no positive signal was toted for the negative controls in the study. The sensitivity of LAMP assay reached up to 2-3 T. gondii tachyzoites/ml per reaction. Conclusion LAMP assay shows proper specificity and sensitivity for the detection of T. gondii.

To localize and characterize the hypothetical protein CT358 in the chlamydial infected cells.CT358 gene from the Chlamydia trachomatis(C.trachomatis) serovar D genome was amplified and cloned into the pGEX and pDSRedC1 vectors.The recombinant plasmid pGEX-CT358 was constructed and expressed as GST fusion proteins.The GST-CT358 fusion protein was used to immunize mice to raise the antibodies,which specifically recognized CT358 without cross-reacting with other unrelated proteins.The antibodies were then used to localize the endogenous CT358 protein and determine the expression pattern in Chlamydial infected cells using an indirect immunofluorescence assay(IFA).Meanwhile,pDSRedC1-CT358 recombinant plasmid was transfected to HeLa cells to evaluate the effect of CT358 expression on the subsequent chlamydial infection.The hypothetical protein CT358 was identified in the inclusion membrane of C.trachomatis-infected cells for the first time,and it was detected as early as 12 h after C.trachomatis infection and remained in the inclusion membrane throughout the rest of the infection cycle.Cytosolic expression of CT358 via a transgene failed to affect the subsequent chlamydial infection.These observations together have demonstrated that CT358 is a newly identified chlamydial inclusion membrane protein,giving the potentially importance for further understanding the mechanisms of chlamydial intracellular parasitism.

OBJECTIVE:To observe the effect of Sanhuang decoction combined with compound amino acid liposome healing membrane on treatment stage Ⅲ pressure ulcer. METHODS:According to the random number table,ninety cases of stage Ⅲ pres-sure ulcer were divided into control group A,control group B,and experimental group with 30 cases in each group. On the basis of the basic processing for each group,control group A was treated by dressing change with Sanhuang decoction only,control group B by dressing change with compound amino acid liposome healing membrane only,and experimental group by dressing change with Sanhuang decoction combined with compound amino acid liposome healing membrane. The results of treatment were observed and the curative effects were compared among 3 groups after 21 days or pressure ulcer healing. Three groups were com-pared with chi-square test and each two groups were compared with chi-square segmentation method. RESULTS:There was no dif-ference on total curative effect in control group A(62.1%)and control group B(66.7%)(P>0.05),however,there was statistical significance on total curative effect in experimental group(93.3%)and control group A(P<0.05),also in experimental group and control group B(P<0.05). CONCLUSIONS:The effect of Sanhuang decoction combined with compound amino acid liposome healing membrane on treatment of stage Ⅲ pressure ulcer is better than the effect of dressing change with Sanhuang decoction only or compound amino acid liposome healing membrane only,and the healing time of wound is shortened. This research is deserved further study.

To localize and characterize the hypothetical protein CT358 in the chlamydial infected cells. CT358 gene from the Chlamydia trachomatis (C. trachomatis) serovar D genome was amplified and cloned into the pGEX and pDSRedCI vectors. The recombinant plasmid pGEX-CT358 was constructed and expressed as GST fusion proteins. The GST-CT358 fusion protein was used to immunize mice to raise the antibodies, which specifically recognized CT358 without eross-reacting with other unrelated proteins. The antibodies were then used to localize the endogenous CT358 protein and determine the expression pattern in Chlamydial infected cells using an indirect immunofluorescence assay (IFA). Meanwhile, pDSRedC 1-CT358 recombinant plasmid was transfected to HeLa cells to evaluate the effect of CT358 expression on the subsequent chlamydial infection. The hypothetical protein CT358 was identified in the inclusion membrane of C. trachomatis-infected cells for the first time,and it was detected as early as 12 h after C. trachomatis infection and remained in the inclusion membrane throughout the rest of the infection cycle. Cytosolic expression of CT358 via a transgene failed to affect the subsequent ehlamydial infection. These observations together have demonstrated that CT358 is a newly identified chlamydial inclusion membrane protein, giving the potentially importance for further understanding the mechanisms of chlamydial intracellular parasitism.

ObjectiveTo purify and characterize the monoclonal antibody (McAb) against Chlamydia trachomatis pORF5 plasmid protein.Methods The hybridoma cells stably secreting specific McAb against pORF5 were cultured in a large scale,and protein G purification by affinity chromatography was used to purify 2H4 McAb.ELISA was used to determine the antibody titer,and identify McAb isotype.Immunofluorescence assay (IFA) and Western blot were performed to detect McAb specificity.Results The purity of 2H4 antibody was 93％,the titer reached 1:1024,and 2H4 McAb was identified to belong to IgG2a isotype,2H4 McAb reacted strongly with the GST-pORF5 fusion protein and endogenous pORF5 protein expressed by Chlamydia trachomatis serovar A,D,L2,Chlamydia muridarum ( MoPn ),Chlamydia psittaci 6BC,but not other chlamydial plasmid proteins and Chlamydia pneumoniae(Cpn) AR39 strain.Conclusion2H4 McAb against pORF5 protein was successfully purified with a high titer and specificity which lay a foundation for further study on pORF5 protein structure and function.

Objective To investigate the cachexia morbidity among hospitalized patients with digestive system cancer and evaluate its impact on clinical outcomes.Method By analyzing the clinical data of 5 118 hospitalized patients with digestive system cancer in Zhongshan Hospital,Fudan University from January 2012 to December 2013,we investigated the cachexia morbidity and compared the clinical outcome between cachectic patients and noncachectic patients.Results The overall cachexia morbidity of hospitalized patients with digestive system cancer was 15.7％ (803/5 118).The highest cachexia morbidity was 34.0％ (89/173),found in patients with pancreatic cancer.In cachectic group and non-cachectic group,the overall completion rate of radical resection was 67.1％ (539/803) and 74.5％ (3 214/4 315),respectively (P =0.000).Compared to the non-cachectic group,the cachetic group had significantly longer postoperative hospital days [(11.5 ±6.2) d vs (9.4 ±4.9) d,P =0.003],slower postoperative recovery of bowel function [(3.4 ±0.9) d vs (3.2 ±0.8) d,P =0.013],longer postoperative time to intake semifluid [(4.4 ± 1.5) d vs (3.9 ± 1.3) d,P =0.002],and more postoperative complications in 28 days after surgery [8.9％ (48/539) vs 5.8％ (186/3 214),P=0.006].After surgery,131 patients in the cachectic group were transferred to the ICU,and 646 patients in non-cachectic group transferred to the ICU (24.3％ vs 20.0％,P=0.026).Compared to the non-cachecic group,the reoperation rate [3.2％ (17/539) vs 1.5％ (48/3214)],ventilator support rate [8.0％ (43/539) vs 5.7％ (184/3 214)],and mortality [2.4％ (13/539) vs 1.1％ (35/3 214)] of the cachectic group were all significantly higher (P =0.006,0.042,0.011).Conclusions Cachexia is common in hospitalized patients with digestive system cancer,especially in patients with pancreatic cancer.Cachexia has negative impact on the clinical outcomes.