OBJECTIVE To explore the clinical characteristics and risk factors for surgical site infections(SSI)in the patients undergoing type Ⅰ incision surgeries of grade Ⅲ and Ⅳ so as to provide theoretical bases for effective prevention and control of SSI.METHODS The clinical data were prospectively collected from 280 patients with SSI who received major type Ⅰ incision surgeries of grade Ⅲ and Ⅳ in the key departments of the Second Affiliated Hospital of Jiaxing University from Jan.2021 to Jun.2024,and randomly from another 280 patients without infec-tions.The enrolled patients were assigned as the infection group and the no infection group,respectively.The clin-ical characteristics of the SSI were analyzed.The baseline data,including sex,age,hypertension,diabetes melli-tus,hyperlipidemia,heart disease,hepatorenal function,nutrition status,preoperative length of hospital stay,operation duration,implantation materials,emergency surgery,intraoperative warm maintenance,microscope,blood loss volume,blood transfusion and postoperative intensive care unit(ICU)stay as well as severe hypopro-teinemia,were collected from the enrolled patients.Univariate analysis and logistic regression analysis were per-formed for the influencing factors for the SSI.RESULTS A total of 16,706 patients underwent neurosurgery,or-thopedics and cardiothoracic surgery,280 of whom had SSI,the incidence of SSI was 1.68％.Totally 264 strains of bacteria were isolated,53.03％of which were gram-positive bacteria,46.97％were gram-negative bacteria;Staphylococcus aureus was the most common species of pathogenic bacteria.The result of multivariate logistic a-nalysis showed that the nutrition status,operation duration,microscope,postoperative ICU and severe hypopro-teinemia were the risk factors for the SSI(P＜0.05).CONCLUSIONS The SSI is common among the patients un-dergoing type Ⅰ incision surgeries of grade Ⅲ and Ⅳ.It is necessary for the hospital to take targeted interven-tion measures in early stage based on the risk factors.

OBJECTIVE To explore the clinical characteristics and risk factors for surgical site infections(SSI)in the patients undergoing type Ⅰ incision surgeries of grade Ⅲ and Ⅳ so as to provide theoretical bases for effective prevention and control of SSI.METHODS The clinical data were prospectively collected from 280 patients with SSI who received major type Ⅰ incision surgeries of grade Ⅲ and Ⅳ in the key departments of the Second Affiliated Hospital of Jiaxing University from Jan.2021 to Jun.2024,and randomly from another 280 patients without infec-tions.The enrolled patients were assigned as the infection group and the no infection group,respectively.The clin-ical characteristics of the SSI were analyzed.The baseline data,including sex,age,hypertension,diabetes melli-tus,hyperlipidemia,heart disease,hepatorenal function,nutrition status,preoperative length of hospital stay,operation duration,implantation materials,emergency surgery,intraoperative warm maintenance,microscope,blood loss volume,blood transfusion and postoperative intensive care unit(ICU)stay as well as severe hypopro-teinemia,were collected from the enrolled patients.Univariate analysis and logistic regression analysis were per-formed for the influencing factors for the SSI.RESULTS A total of 16,706 patients underwent neurosurgery,or-thopedics and cardiothoracic surgery,280 of whom had SSI,the incidence of SSI was 1.68％.Totally 264 strains of bacteria were isolated,53.03％of which were gram-positive bacteria,46.97％were gram-negative bacteria;Staphylococcus aureus was the most common species of pathogenic bacteria.The result of multivariate logistic a-nalysis showed that the nutrition status,operation duration,microscope,postoperative ICU and severe hypopro-teinemia were the risk factors for the SSI(P＜0.05).CONCLUSIONS The SSI is common among the patients un-dergoing type Ⅰ incision surgeries of grade Ⅲ and Ⅳ.It is necessary for the hospital to take targeted interven-tion measures in early stage based on the risk factors.

Bacteremia induced by periodontal infection is an important factor for periodontitis to threaten general health. P. gingivalis DNA/virulence factors have been found in the brain tissues from patients with Alzheimer's disease (AD). The blood-brain barrier (BBB) is essential for keeping toxic substances from entering brain tissues. However, the effect of P. gingivalis bacteremia on BBB permeability and its underlying mechanism remains unclear. In the present study, rats were injected by tail vein with P. gingivalis three times a week for eight weeks to induce bacteremia. An in vitro BBB model infected with P. gingivalis was also established. We found that the infiltration of Evans blue dye and Albumin protein deposition in the rat brain tissues were increased in the rat brain tissues with P. gingivalis bacteremia and P. gingivalis could pass through the in vitro BBB model. Caveolae were detected after P. gingivalis infection in BMECs both in vivo and in vitro. Caveolin-1 (Cav-1) expression was enhanced after P. gingivalis infection. Downregulation of Cav-1 rescued P. gingivalis-enhanced BMECs permeability. We further found P. gingivalis-gingipain could be colocalized with Cav-1 and the strong hydrogen bonding between Cav-1 and arg-specific-gingipain (RgpA) were detected. Moreover, P. gingivalis significantly inhibited the major facilitator superfamily domain containing 2a (Mfsd2a) expression. Mfsd2a overexpression reversed P. gingivalis-increased BMECs permeability and Cav-1 expression. These results revealed that Mfsd2a/Cav-1 mediated transcytosis is a key pathway governing BBB BMECs permeability induced by P. gingivalis, which may contribute to P. gingivalis/virulence factors entrance and the subsequent neurological impairments.
Animals ; Rats ; Bacteremia/metabolism* ; Blood-Brain Barrier/microbiology* ; Caveolin 1/metabolism* ; Gingipain Cysteine Endopeptidases/metabolism* ; Permeability ; Porphyromonas gingivalis/pathogenicity* ; Transcytosis ; Virulence Factors/metabolism*

Aim To explore the anti-apoptotic function of cardiac progenitor cells(CPCs)-derived exosome in vitro.Method CPCs were isolated from mouse heart using Magnetic Cell Sorting(MACS)system.Flow Cy-tometry(FC)determine the purity of stem cell surface antigen-1 positive(Sca-1 +)CPCs.Exosome was puri-fied from conditional medium,and confirmed by West-ern blot using CD63 as a marker,Nanoparticle Traffic-king Analysis(NTA)was used to detect the diameters and concentration of exosome.Then the cells were di-vided into control groups and CPC-exosome pre-protec-tion groups.H2 O2 was added into H9c2 cells to induce oxidative stress.Western blot was adopted to determine the expression of cleaved caspase-3.Results ① Im-munofluorescence showed that CPCs isolated by MACS were positively expressing Sca-1 protein;FC analysis showed that typical purity of Sca-1 +CPCs from the first
preparations was more than 95%.② WB demonstrated that CD63 of exosome isolated from CCMwas positively expressed,and NTA results showed that the diameters of exosome were (82.33 ±3.06)nm(n =3).Micro-scope detected PKH-26 labeled exosome appeared in the cytoplasma of H9c2 cells.③ Western blot showed the CPC-exosome pre-protection groups significantly down-regulated the levels of cleaved caspase-3 com-pared to the control groups(P <0.05).Conclusion CPC can secrete exosome which carries many important cargos,which can effectively gather in H9c2 cells. CPC-exosome can protect H9c2 cells from the oxidative stress induced by H2 O2 .Our results highlight a new perspective strategy for cardiac disease.

In this study ,the objective is to establish a nested‐PCR assay for the detection of H .bilis with high sensitivity and specificity .The nested primers were designed based on sequences of 16S rRNA gene of seventeen subtypes of H .bilis .Af‐ter optimizing reaction condition ,the sensitivity and specificity of the assay were examined via the detection of feces simulated samples ,mice infection model samples and clinic patients’ samples .The detection sensitivity of H .bilis strain for feces simu‐lated samples was 10 CFU/100 μL .H .bilis was successfully detected in the liver ,caecum and feces of experimentally infected mice .Moreover ,H .bilis was successfully detected in the bile ,cholecyst mucous membrane and feces samples from two of ten patients with cholelithiasis .Due to the PCR assay’s high sensitivity and specificity ,the method may be used to detect the infec‐tion of H .bilis .

Objective To establish ultra-high performance liquid chromatography-evaporative light scattering detector (UPLC-ELSD) method to determine the content of astragaloside in Guiqi Yiqi Yangxue Mixture and optimize the best water decoction process of the mixture.Methods The UPLC-ELSD conditions to determine the content of astragaloside in the mixture were as follow.The method was carried out on a Waters Acquity UPLC BEH C18 column (2.1 mm ×50 mm,1.7 μm) by using acetonitrile-water (35∶65) as mobile phase, and the flow rate was 0.3 mL/min and the column temperature 30℃.The parameter of ELSD:the drift tube temperature was set at 60℃, and the gas flow rate of N2 2.0 L/min.The content of paeoniflorin in the mixture was detected by previously established HPLC method.The orthogonal test was designed to optimize the water decoction process of the amount of water (A), decoction time (B) and decoction times (C), and the extract yield and the contents of astragaloside and paeoniflorin were evaluated as the comprehensive markers for the purpose.Results The UPLC-ELSD determination of astragaloside had a good linear relationship (r =0.9999), its mean recovery was 103.0%, and it had a good precision, specificity and repeatability.The orthogonal experimental results showed that decoction times (C) >the amount of water (A) >decoction time (B) were major factors affecting the decoction process in order and the ANOVA result showed that decoction times has significant effect on the decoction process (P<0.05).The optimal process of Guiqi Yiqi Yangxue Mixture was adding eight-fold water each time to decoct twice for 1.5 h totally.Conclusion This optimized water decoction process is feasible and stable, and suitable for industrialized production.

Objective To investigate the diagnosis and treatment of ectopic para-aortic pheochromocytoma. Methods Clinical data of 2 cases with ectopic para-aortic pheochromocytoma in the Third Affiliated Hospital of Sun Yat-sen University from August 2012 to December 2013 were analyzed retrospectively. The informed consents of both patients were obtained and local ethical committee approval had been received. Both patients were female with the age of 54, 57 years old. The occurrence of disease, diagnosis, treatments and efficacy were observed. Results Case 1 was admitted in hospital for complaint of 1-month distending pain in upper abdominal and ifnding of retroperitoneal space-occupying lesion for 8 d. Solid space-occupying lesion at the right side of mid-upper abdomen was found by abdominal ultrasound and computer tomography (CT) examination. The preoperative diagnosis was abdominal mesenchymoma. Case 2 was admitted in hospital for complaint of palpitation, chest pain and ifnding of hepatic space-occupying lesion for 10 d and had a history of 5-year hypertension. Cystic solid space-occupying lesion in the hepatic caudate lobe was found by abdominal ultrasound, CT and magnetic resonance imaging (MRI) examinations and cystadenoma was suspected. The 2 patients underwent resection of ectopic para-aortic pheochromocytoma under general anesthesia by tracheal intubation. The intraoperative exploration found that touching the tumor had great impacts on blood pressure and heart rate and ectopic para-aortic pheochromocytoma was diagnosed. The patients were transferred to surgical intensive care unit (ICU) for monitoring and treatment. Pheochromocytoma was confirmed by the pathological examination. The patients were recovered and discharged from hospital and remained well till the paper submission date. Conclusions Ectopic para-aortic pheochromocytoma is a rare disease without characteristic clinical features. It is dififcult for preoperative diagnosis. The conifrmation is mainly depends on the pathological ifndings. Surgical resection is the priority of treatment. Fully assessment of operative risk before operation, avoiding touching tumor tissues during operation, complete excision of tumor capsule, close monitoring on the blood pressure after operation can achieve a good clinical treatment outcome.

Human cytomegalovirus glycoprotein complex Ⅱ (gC Ⅱ ) consists of two glycoproteins, gM and gN. Although gC Ⅱ specific IgG purified from HCMV positive patient sera can neutralize HCMV, there has been no report on the generation of virus-neutralizing antibodies by immunizing with one epitope of gM. The epitope, termed MAD, was screened from random phage peptide library by subtractive strategy. The peptide sequence of MAD was highly homologous with 32~38 amino acids of HCMV gM. Mice immunized with MAD coupled with keyhole limpet hemocyanin (KLH) could produce specific antibodies against MAD, and the antibodies obtained could bind not only native HCMV particles, but also the recombinant gM30～78 peptide. ELISA analysis results showed that MAD could specifically bind HCMV-positive human serum samples. Virus-neutralizing assay results demonstrated that the antibodies against MAD could inhibit HCMV strain AD169 entering the human embryonic lung cells. The results suggested that MAD could be used as a new potential protective antigen in the development of HCMV vaccine.

OBJECTIVETo search for the dystrophin gene mutations of Duchenne muscular dystrophy (DMD) patients without gross deletions, in order to offer accurate genetic counseling and prenatal diagnosis for those families.

METHODSAll 79 exons of the dystrophin gene as well as its 5'-UTR and 3'-UTR of 14 Chinese DMD/Becker muscular dystrphy (BMD) patients without detectable gross deletions were screened by denaturing high performance liquid chromatography (DHPLC) and heteroduplex fragments were identified by subsequent sequencing.

RESULTSSeven causative point mutations, including two novel ones, were detected in 7 patients. Fourteen known polymorphisms and 7 unknown intronic variations were also detected. Five mothers of the patients were obligate carriers.

CONCLUSIONDHPLC is an efficient way of identifying point mutations and the female carriers in DMD families.

Adolescent ; Asian Continental Ancestry Group ; genetics ; Base Sequence ; Child ; Child, Preschool ; DNA Mutational Analysis ; Dystrophin ; genetics ; Exons ; genetics ; Female ; Genetic Counseling ; Genetic Testing ; methods ; Humans ; Introns ; genetics ; Male ; Muscular Dystrophy, Duchenne ; diagnosis ; genetics ; Point Mutation ; Polymorphism, Genetic ; Pregnancy ; Prenatal Diagnosis ; Sequence Deletion ; genetics

Objective To isolate and identify the potential binding partners of LRRK2,a gene linked to both dominant familial form and sporadic form of Parkinson's disease,thus to further our knowledge of its function.Methods We used a sequence containing full-length of COR domain and part of ROC and MAPKKK domain as bait.The bait amplified by polymerase chain reaction(PCR) was then cloned into a yeast expression plasmid pGBKT7.After being sequenced and analyzed,pGBKT7-bait was transformed into the yeast strain AH109.Western blot was performed to confirm the expression of pGBKT7-bait in AH109 yeast strain.Then human fetal brain cDNA library was trarnsformed into that yeast strain.which could express pGBKT7-bait fusion protein.The yeast strain which contained pGBKT7-bait and human fetal brain cDNA library was plated on quadruple dropout medium (SD/-Trp/-Leu/-His/-Ade)containing X-a-gal.We retested these positive colonies using 2 independent yeast strains AH109 contained pGBKT7-bait or pGBKT7,respectively.At last,these plasmids isolated from these true positive colonies were analyzed by bioinformatics.Results We obtained 9 true positive colonies,these colonies were sequenced, and we performed sequence Blast in GenBank.Three colonies of the 9 positive colonies were not in open reading-frames.Among other 6 colonies,there were known proteins including spermatid perinuclear RNA-binding protein(STRBP)and BCL2-associated athanogene 5 isoform b(BAG5),as well as unknown proteins including tyrosine phosphatase non-receptor type(PTPN23),1(3)mbt-like 3 isoform b(L3 MBTL3),RALY RNA binding protein-like isoform 1(RALYL),and Homo sapiens mRNA for KIAA1783 protein,partial cds(KIAA 1783).Conclusion True positive colonies of LRRK2 are successfully obtained by the yeast 2-hybrid.Our screened proteins may provide a new research clue for revealing biological functions of LRRK2,pathogenesis of Parkinson's disease,and other neurodegerations.