Objective: To facilitate the functional analysis of chromosomal genes and their products, the recombineering technique to epitope tagging of chromosomal genes of Y. pestis was adapted. Methods: The epitope tag was generated by primer annealing and then fused with resistance gene by fusion PCR. The epitope-resistance cassette was inserted into pBluecript, resulted in the template plasmid, pBS-MH. The tagging cassette for rpoS was obtained by PCR amplification from pBS-MH with primers containing homology specific to the target gene. PCR products were transformed into recombination competent cells and recombinants were selected. PCR and DNA sequencing were used to confirm the correct tagging event. The expression of the tagged protein was detected with Western blot by using monoclonal antibody to the epitope. Results: The template plasmid containing fusion of epitope and resistance gene was successfully constructed. The sigma factor gene, rpoS, was tagged with a myc-his tag at the COOH terminus. Expression of the tagged rpoS was successfully detected indirectly by the antibody against His tag. Conclusion: The chromosomal gene tagging by recombineering technique represents a powerful tool in the functional study of bacterial genes and their products.

Aged ; Aged, 80 and over ; Aging ; Female ; Fracture Fixation, Intramedullary ; methods ; Hip Fractures ; pathology ; physiopathology ; surgery ; therapy ; Humans ; Male ; Middle Aged ; Minimally Invasive Surgical Procedures ; methods ; Reconstructive Surgical Procedures ; methods ; Recovery of Function ; Treatment Outcome

OBJECTIVETo investigate the expression of Csk-binding protein (CBP) in esophageal carcinoma and its association with the tumorigenesis and progression of esophageal cancer.

METHODSRT-PCR and Western blotting were employed to determine the expressions of CBP at the mRNA and protein levels in 50 pairs of fresh esophageal carcinoma tissue and the adjacent normal tissues.

RESULTSCBP mRNA and protein expressions in normal tissues were 1.43- and 1.28-fold higher than those in the cancer tissues, respectively (P<0.05). The expressions of CBP mRNA and protein were positively correlated (P=0.015). The decreased expressions of CBP were significantly correlated to lymph node metastasis of esophageal cancer (P<0.05).

CONCLUSIONThe expression of CBP gene is decreased in esophageal carcinoma, which might contribute to the tumorigenesis and progression of this malignancy.

Adenocarcinoma ; metabolism ; pathology ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; metabolism ; pathology ; Down-Regulation ; Esophageal Neoplasms ; metabolism ; pathology ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; src-Family Kinases ; genetics ; metabolism

OBJECTIVETo investigate the effects of nucleoside analogues on hepatitis B virus (HBV) in hepatic lymph nodes of hepatitis B related liver transplantation recipients who were hepatitis B surface antigen (HBsAg) positive but negative for serum HBV DNA.

METHODSFrom June 2010 to March 2011, thirty-six cases of hepatitis B related liver transplantation recipients [32 males, 4 females, average age (54 ± 7) years] were divided into drug treatment group and non-drug treatment group according to the utility of nucleoside analogues. Drug treatment group was divided into two subgroups: drug treatment > 3 months group and drug treatment ≤ 3 months group. The hepatic lymph nodes in the hepatoduodenal ligament were taken during the operation of liver transplant. Using nested or semi-nested PCR, HBV DNA and the replicative form HBV cccDNA in hepatic lymph nodes were detected. Data were analyzed by Fisher's exact test.

RESULTSThe positive rate of HBV DNA: the difference was not statistically significant between drug treatment group (72.7%, 16/22) and non-drug treatment group (14/14) (P = 0.062), the difference was not statistically significant between drug treatment > 3 months group (10/14) and drug treatment ≤ 3 months group (6/8) in the subgroups of drug treatment group (P = 1.000). The positive rate of HBV cccDNA: drug treatment group (22.7%, 5/22) was significantly lower than the non-drug treatment (12/14) (P = 0.000), drug treatment > 3 months group (1/14) was significantly lower than drug treatment ≤ 3 months group (4/8) in the subgroups of drug treatment group (P = 0.039).

CONCLUSIONSHepatic lymph nodes maybe one of the extrahepatic HBV reservoirs. Treating with nucleoside analogues more than 3 months can significantly decrease the replication of HBV in hepatic lymph nodes of HBV associated liver transplantation recipients.

Adult ; Aged ; DNA, Viral ; analysis ; Female ; Hepatitis B ; drug therapy ; Hepatitis B virus ; drug effects ; physiology ; Humans ; Liver Transplantation ; Lymph Nodes ; virology ; Male ; Middle Aged ; Nucleosides ; therapeutic use ; Preoperative Care ; Virus Replication

Background:BK polyomavirus (BKPyV)-associated nephropathy (BKPyVAN) is an important cause of dysfunction and failure of renal transplants.This study aimed to assess the diagnostic performance of morning urine specific gravity (MUSG) in diagnosing BKPyVAN in kidney transplant recipients.Methods:A total of 87 patients,including 27 with BKPyVAN,22 with isolated BKPyV viruria,18 with T cell-mediated rejection (TCMR),and 20 with stable graft function,were enrolled in the First Affiliated Hospital of Sun Yat-Sen University from March 2015 to February 2017.MUSG at biopsy and during a follow-up period of 24 months after biopsy was collected and analyzed.Receiver operating characteristic (ROC) curve analysis was used to determine the ability of MUSG to discriminate BKPyVAN.Results:At biopsy,the MUSG of BKPyVAN group (1.008 ± 0.003) was significantly lower than that of isolated BK viruria group (1.013 ± 0.004,P ＜ 0.001),TCMR group (1.011 ± 0.003,P =0.027),and control group (1.014 ± 0.006,P ＜ 0.001).There was no significant difference in MUSG among the isolated BK viruria group,TCMR group,and control group (P =0.253).In BKPyVAN group,the timing and trend of MUSG elevate were consistent with the timing and trend of the decline of viral load in urine and plasma,reaching a statistical difference at 3 months after treatment (1.012 ± 0.003,P ＜ 0.001) compared with values at diagnosis.ROC analysis indicated that the optimal cut-off value of MUSG for diagnosis of BKPyVAN was 1.009,with an area under the ROC curve (AUC) of 0.803 (95％ confidence interval [CI]:0.721-0.937).For differentiating BKPyVAN and TCMR,the optimal MUSG cut-off value was 1.010,with an AUC of 0.811 (95％ CI:0.687-0.934).Condusion:Combined detection of MUSG and BKPyV viruria is valuable for predicting BKPyVAN and distinguishing BKPyVAN from TCMR in renal transplant recipients.

OBJECTIVETo compare the efficiency of response evaluation by clinical examination, ultrasonograghy and mammography in neoadjuvant chemotherapy (NAC) for breast cancer.

METHODSA retrospective cohort study was conducted to analyze the data of 141 patients treated with neoadjuvant chemotherapy. Response evaluation was performed by clinical palpation, ultrasound and mammography.

RESULTSOnly 12 (8.5%) among the 141 patients presented with a stage I tumor. The tumor size determined by palpation was often larger than that by ultrasound before therapy (P < 0.01). Among patients with suspicions axillary nodes checked by ultrasound, 88.3% (53/60) of them had positive nodes by pathology before NAC, and 34.5% (10/29) of patients with negative nodes determined by ultrasound had positive nodes by pathology. In all the 141 patients, 21(14.9%) showed pathological complete remission in both the primary tumor and lymph node. For response evaluation, the false complete remission rate judged by clinical examination was 46.8% (22/47), and the false tumor residual rate by ultrasound was 84.0% (21/25). In 53.5% (23/43) of patients the response could not be assessed by mammography due to that the tumors were undistinguishable in size. The range of microcalcification was not reduced in 5 patients with a partial response of the tumor. 25 patients experienced needle puncture during therapy. Among them, in the 9 pathologically negative patients, only 3 achieved pCR, and the other 16 positive patients didn't achieve pCR.

CONCLUSIONUsing the puncture or sentinel lymph node biopsy, clinicians should pay enough emphasis on the pathological determination of the node status before chemotherapy. Clinicians will make a quite of false judgment of the tumor by clinical examination, ultrasound or mammography. They may use needle puncture during therapy to evaluate the response of neoadjuvant chemotherapy, and the result should be analyzed synthetically.

Adult ; Aged ; Aged, 80 and over ; Axilla ; Breast Neoplasms ; diagnostic imaging ; drug therapy ; pathology ; Carcinoma, Ductal, Breast ; diagnostic imaging ; drug therapy ; pathology ; Chemotherapy, Adjuvant ; Cohort Studies ; Female ; Humans ; Lymph Nodes ; pathology ; Lymphatic Metastasis ; Mammography ; Middle Aged ; Neoadjuvant Therapy ; Neoplasm Staging ; Remission Induction ; methods ; Retrospective Studies ; Sentinel Lymph Node Biopsy ; Ultrasonography

OBJECTIVE: To explore the different brain areas activated by Chinese, English, and Nepali word tasks in Nepalese by using the functional magnetic resonance image (fMRI). METHODS: To determine the neuroanatomic functional brain areas responsible for Chinese, English and Nepali reading as well as sentence-formation, blood oxygenation level dependent (BOLD) block design by fMRI was performed in 6 healthy Nepalese volunteers. RESULTS: During Chinese reading, the activated areas included bilateral motor area, subfrontal gyri, superior temporal gyri, and superior parietal lobule; during English reading, the activated areas were left motor area, left subfrontal gyrus, left supra temporal gyrus, left insula and bilateral cerebellum; and the Nepali task demonstrated the activation of left anterior central gyrus, superior frontal gyrus, superior temporal gyrus. Aside from the bilateral occipital lobes, both English and Nepali activated areas were the left cerebral hemisphere dominant. CONCLUSION The more familiar with the language, the fewer areas are activated. Superior temporal gyrus might be involved in sentence-formation.
Adult ; Brain ; physiology ; China ; England ; Female ; Humans ; Image Processing, Computer-Assisted ; Language ; Magnetic Resonance Imaging ; Nepal ; Pattern Recognition, Visual ; Reading

Objective To compare the clinical efficacy of orthotopic ileal neobladder versus ortho- topic sigmoid neobladder.Methods The data of 96 patients who had undergone orthotopic ileal neoblad- der and 68 patients who had undergone orthotopic sigmoid neobladder were retrospectively analyzed.The perioperative condition,urinary continence,urodynamics,and pouch-related complications were compared between the 2 groups.Results Of all the 164 patients,12(7.3%)were lost to follow-up.The mean fol- low-up was 46(2-86)months in orthotopic ileal neobladder group,and 42(4-78)months in orthotopic sigmoid neobladder group.There was no significant difference in intraoperative blood loss and postoperative urinary continence between the 2 approaches(P＞0.05).However,compared with sigmoid neobladder group,ileal neobladder group had longer operative time and postoperative recovery time,and got a bigger pouch(P＜0.05).The early and late pouch-related complication rates of ileal neohladder group were 16. 7% and 29.2%,which were higher than those of sigmoid neobladder group.During the follow-up,tumor recurred in 3 cases of ileal neobladder group,but none in sigmoid neobladder group.Conclusions Ortho- topic ileal neobladder and sigmoid neobladder are similar in operative difficulties,and both can achieve satis- factory clinical results.Compared with ileal neobladder,sigmoid neobladder has shorter operative time, quicker recovery and lower rate of pouch-related complications,thus is a preferred procedure.

Objective To analyze the evolution and genetic characterization of echovirus 11 (Echo11 ) from the acute flaccid paralysis (AFP) cases in Shandong province. Methods Isolation of Enterovirus was performed from stool samples of AFP cases from 1994 to 2009. All positive strains were sero-typed by neutralization test. Entire VP1 coding region from 27 strains typed as Echo 11 was amplified by reverse transcription-polymerase chain reaction(RT-PCR), and subsequently phylogenetic analyse on VP1 sequences from these strains and others published in GenBank were conducted. Results From 1994 to 2009, altogether 915 non-polio enterovirus (NPEV) strains were isolated with 79(8.6% ) isolates serotyped as Echo11. There were 876 nucleotides (nt) in the complete VP1 genes, encoding 292 amino acids (aa). The nt identities were 76.4%-100.0% among those Echo11 Shandong strains with the aa identities as 91.4% -100.0%. The nt and aa identities were 77.7%-80.7% and 90.7%-94.8% between Shandong strains and prototype strains, respectively.Conclusion All Echo11 strains could be divided into four genotypes. Shandong strains that forming three (A1, A2 and C1) new sub-genotypes, with every sub-genotype had several brands.Sub-genotype A1 appeared to be the lately circulating one.

OBJECTIVETo investigate the expression of oncoprotein c-erbB2 in primary breast cancer and to analyze its relation to its prognosis.

METHODSImmunohistochemical staining for c-erbB2 was performed on paraffin-embedded specimens of primary breast cancer from 284 patients, and the relation to its prognosis was statistically analyzed.

RESULTSPositive expression rate of c-erbB2 was 26.8% (76/284) in 284 primary breast cancer patients. Expression of c-erbB2 was positively correlated with the status of lymph node metastasis (P = 0.003). Univariate analysis indicated that c-erbB2 expression is a significant prognostic factor for the disease-free survival (DFS) (P = 0.024) and overall survival (OS) (P = 0.002), while multivariate analysis demonstrated that c-erbB2 is an independent prognostic factor for OS (P = 0.023). Moreover, tumors with c-erbB2 positive expression are more tend to metastasis to other viscera than those with c-erbB2 negative. c-erbB2 expression has different prognostic values for patients with different status of estrogen receptor (ER) and lymph node metastasis.

CONCLUSIONc-erbB2 expression is an independent prognostic factor for total survival time in primary breast cancer patients, and its prognostic values are different according to the different ER status and lymph node metastasis.

Adult ; Aged ; Breast Neoplasms ; metabolism ; pathology ; therapy ; Carcinoma, Ductal, Breast ; metabolism ; pathology ; therapy ; Chemotherapy, Adjuvant ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Mastectomy, Radical ; Middle Aged ; Prognosis ; Proportional Hazards Models ; Radiotherapy, Adjuvant ; Receptor, ErbB-2 ; metabolism ; Receptors, Estrogen ; metabolism ; Survival Rate