Objective:To evaluate the clinical efficacy of intra-biliary drainage versus T-tube drainage following laparoscopic common bile duct exploration (LCBDE) for choledocholithiasis.Methods:The clinical data of 50 patients undergoing LCBDE for choledocholithiasis in Dalian Friendship Hospital of Dalian Medical University from January 2018 to October 2022 were retrospectively analyzed, including 23 males and 27 females, aged (61.3±16.2) years old. Patients were divided into the intra-biliary drainage group and T-tube drainage group. Propensity score matching was used to match the baseline data of the two groups at a 1∶1 ratio. The operation time, intraoperative blood loss, postoperative hospital stay, abdominal drainage tube indwelling time, postoperative bile drainage volume and postoperative complications were compared between the groups.Results:Compared with the T-tube group, the operative time [(155.0±36.5) min vs. (194.4±55.8) min], length of postoperative hospital stay [8.0(7.0, 8.0) d vs. 11.0(8.0, 13.0) d], and abdominal drainage tube indwelling time [5.0(4.0, 6.0) d vs. 6.0(5.0, 8.0) d] were all shorter in the intra-biliary drainage tube group (all P<0.05). The postoperative bile drainage volume was reduced [0 ml vs. 431.4(344.7, 484.3) ml]. No postoperative bile leakage occurred in either group. The intraoperative blood loss, proportion of postoperative residual stone, stone recurrence and biliary stricture were comparable between the two groups (all P>0.05). Conclusion:Intra-biliary tube drainage following LCBDE could be safe and effective for choledocholithiasis. Compared to the classic procedure of T-tube drainage, it may be superior in the operation time, postoperative hospital stay, abdominal drainage tube indwelling time, postoperative bile drainage volume.

OBJECTIVE To investigate the effects of nasal administration of non-mitogenic acid fibroblast growth factor(NMFGF1) loaded nanoparticles(NMFGF1-NPs) on the improvement of cognitive function in vascular dementia(VD) mice and its mechanism. METHODS Nanoparticles containing NMFGF1(NMFGF1-NPs) were prepared by water-in-water emulsion technique and characterized. The mice were divided into sham group, VD model group, blank-NPs group, NMFGF1 solution group and NMFGF1-NPs group after repeated cerebral ischemia-reperfusion to establish VD test model, and then given the corresponding form of drug intervention by nasal cavity. After drug intervention, Morris water maze was used to evaluate the learning and memory function of the animals in each group from the perspective of behavior. Meanwhile, the morphology, arrangement and apoptosis index(AI) of hippocampal neurons in each group were evaluated by pathological methods such as HE staining, FJB staining and Tunel apoptosis staining. In addition, ELISA and Western blotting were used to investigate the molecular mechanism of NMFGF1-NPs improving VD by nasal administration. RESULTS The morphology of NMFGF1-NPs was round. The encapsulation rate of NMFGF1-NPs respectively was (87.76±5.89)%. Morris water maze results showed that the behavioral indexes of mice in VD model group were significantly different from those in sham operation group(P<0.01). At the same time, the pathological results showed that the neurons in the CA1 region of the hippocampus in the VD model group were disordered, the cells morphology and structure were missing, and the AI was significantly increased compared with that in the sham operation group(P<0.01). Meanwhile, compared with the VD model group, the NMFGF1-NPs treatment group showed significant improvement in various behavioral indexes, and the hippocampal neuron cells were intact and orderly, and the AI index was significantly decreased(P<0.01). ELISA and Western blotting analysis showed that compared with that of VD model group and other intervention groups, the content of MDA in the brain of NMFGF1-NPs treatment group was significantly decreased. While the content of SOD, NO and the expressions of Nrf2, SOD-1 and GSTO1/2 was significantly increased (P<0.01). CONCLUSION Nasal administration of NMFGF1-NPs can play the role of antioxidant stress damage by activating Nrf2/ARE signal pathway, and ultimately improve the learning and cognitive function of VD mice.

Background and purpose:The third generation of aromatase inhibitors (AI) in postmenopausal hormone receptor-positive patients is the routine treatments in endocrine therapy. The 500 mg fulvestrant showed clini-cal beneifts in patients with previous AI treatment. This study aimed to access the effcacy and safety of 500 mg fulves-trant in estrogen receptor (ER) positive postmenopausal patients who had previous AI treatments with locally advanced and metastatic breast cancer.Methods:This study retrospectively analyzed the clinical data from 188 post-AI ER positive and (or) progesterone receptor (PR)-positive locally advanced and metastatic breast cancer patients treated with 500 mg fulvestrant in Fudan University Shanghai Cancer Center from Jul. 2011 to Dec. 2015. Primary end point was progression-free survival (PFS). Secondary end points were objective response rate (ORR), clinical beneift rate (CBR) and safety proifle.Results:After the median follow-up of 11.3 months, median PFS was 5.9 months (95%CI: 4.2-7.5), CBR was 40.0% and ORR was 3.4%. COX proportional hazards regression analysis indicated that PFS was correlated with the number of metastatic sites (HR=1.92, 95% CI: 1.2-2.9,P =0.002) and previous lines of chemotherapy (HR=1.52, 95%CI:1.0-2.1,P=0.022). Six patients stopped the treatment for intolerable adverse events.Conclusion:The treatment of 500 mg fulvestrant has a favorable effcacy and safety in treatment of post-AI ER positive postmenopausal patientswith metastatic breast cancer.

Objective To evaluate the effects of TAT-heme oxygenase-1 (TAT-HO-1) fusion protein on liver injury in rats undergoing orthotopic liver transplantation (OLT).Methods Adult male Lewis (inbred) rats (aged 8-10 weeks,weighing 180-230 g) were used as donors and Brown Norway rats (aged 8-10 weeks,weighing 180-230 g) as recipients.The recipient rats were randomly divided into 2 groups (n=6 each) using a random number table:OLT group and TAT-HO-1 group.The livers were harvested according to the method described by Kamada.In OLT group,the donor livers were flushed and preserved with 4 ℃ HTK solution,while the livers were flushed and preserved for 6 h with 4 ℃ HTK solution containing TAT-HO-1 50 μg/ml in group P.Blood samples were obtained at 7 days after transplantation for measurement of activities of alanine aminotransferase and aspartate aminotransferase in serum.Hepatic specimens were obtained at 7 days after transplantation and stained with haematoxylin and eosin for examination under light microscope.Rejection activity index was calculated according to Banff criteria.The contents of transforming growth factor-beta 1 and interleukin-6 in liver tissues were determined using ELISA.Kupffer cells were isolated and cultured for 48 h to determine the levels of transforming growth factor-beta 1 and interleukin-6 in culture medium.Results Activities of alanine aminotransferase and aspartate aminotransferase in serum,rejection activity index and levels of transforming growth factor-beta 1 and interleukin-6 in liver tissues and culture medium of Kupffer cells were significantly decreased,and the pathological changes of livers were mitigated in group TAT-HO-1 as compared to group OLT.Conclusion TAT-HO-1 fusion protein applied during cold storage of donor livers can attenuate liver injury in rats undergoing OLT.

AIM: Aquaporin 3 ( AQP3 ) water channel expressed in the kidney plays a critical role in the urine concentrating mechanism. Mice with AQP3 deletion show a urinary concentrating defect. To better characterize this defect, we studied the influence and mechanism of an acute urea load in conscious AQP3 - null and wild - type mice. METHODS:Urine was collected and assayed every 2 h, from 2 h before (baseline) to 8 h after the urea load. Urine volume, urine osmolality and urea concentration were measured. RNA was isolated from the whole kidney and real - time PCR was carried out using a LightCycler. Total protein of UTs was assayed from kidney tissue by Western blotting. RESULTS: Mice of both genotypes excreted the urea loaded in ～ 4 h with the same time course. Despite their low baseline, the AQP3 - null mice raised their urine osmolality and urea concentration progressively after the urea load to the values almost equal to those in wild - type mice at 8 h. In contrast, urine non - urea solute concentration did not change. Urine volume fell in the last 4 h to about one - fourth of basal values. The urea load strongly upregulated urea transporter UT - A3 expression in both genotype mice. CONCLUSION: These observations show that lack of AQP3 does not interfere with the ability of the kidney to concentrate urea but impairs its ability to concentrate other solutes. This solute - selective response results from the capacity of AQP3 to transport not only water but also urea, suggesting a novel role for AQP3 in non - urea solute concentration in the urine.

BACKGROUND: Cord blood stem cells are one of ideal target cells for gene therapy, but low gene transferring rate is the main difficulty at recent. Janus kinase tyrosine 2 (JAK2) plays an important role in self-renewing of cord blood stem/progenitor cell12s. Therefore, cord blood CD34+ cell line modified by target-amplified JAK2 genes has been developed yet by using gene regulating expression technique in order to overcome low transferring rate of cord blood genes.OBJECTIVE: To investigate the feasibility and reliability of a long-term amplified regulation for cord blood stem/progenitor cells mediated by transgene JAK2. SETTING: Department of Hematology, Beijing Hospital, Ministry of Health.MATERIALS: The experiment was carried out in the Laboratory of Hematological Department, Beijing Hospital, Ministry of Health from June 2003 to April 2006. Cord blood was derived from umbilical cord which was immediately cut from healthy, full-term and natural-parturition infants and was provided by Department of Obstetrics & Gynecology, Beijing Hospital. The experiment was approved by the local ethical committee, and informed consent was obtained from expectant mothers and their relatives for the use of cord blood cells. MiniMACS magnetic separation apparatus and immunomagnetic beads adsorbing CD34 single antibody were provided by Miltenyi Biotec Company, Germany; flow cytometer by FACScalibur, USA; recombinant human stem cell factor (rhSCF), Flt3 ligand (FL), human interleukin-6 (hIL-6), granulocyte macrophage colony stimulating factor (GM-CSF), granulocyte colony-stimulating factor (G-CSF) and thrombopoeitin (TPO) by PeproTec Company; nude mice of the SPF level by Animal Center of Beijing Medical University.METHODS: Retroviral vector MGI-F2JAK2, which was composed of functional catalytic domain of JAK2 genes and two site proteins (2xF36v, F2) combined with synthetic drug (AP20187) of target gene of small molecules, was constructed. AP20187 might specially combine with F36v to cause dimerization of JAK2 so as to activate signal conduction in cells. In addition, the vector included green fluorescence protein reporter gene, which was regarded as a label to detect proliferation. MiniMACS magnetic separation apparatus was used to purify and separate cord blood CD34+ cells. While, retrovirus supernatant including JAK2 was used to transfer cord blood CD34+ cells. After transduction, CD34+ cells were cultured with stem cell factor (SCF), Flt3 ligand, TPO and IL-6 and divided into control group (not adding AP20187) and experimental group (AP20187).MAIN OUTCOME MEASURES: ① Flow cytometer was used to detect percentage of green fluorescence protein reporter gene in the CD34+ cells and to determine gene transferring rate. ② Colony culture results of cord blood stem/progenitor cells after amplification. ③ Nude mice were given subcutaneous injection of ten-week cultured cord blood CD34+ cells at costa and neoplasia was observed after 30 days. RESULTS: ① Plentiful amplification of CD34+ cells was observed in both experimental group and control group. With the culture time passing by, positive rate of gel-filtered platelet of amplified CD34+ cells in the experimental group was gradually increased based on the basic level and more than 95% in the 11th week; however, positive rate of green fluorescence protein reporter gene in the control group was gradually decreased below the basic level and disappeared finally. ② Transgenic CD34+ cells in the experimental group still could generate brust forming unit-erythroid (BFU-E), colony-forming units granulocute/monocyte (CFU-GM) and multipotential hematopoietic progenitors (CFU-Mix); especially, CFU-GM was the main cell in hemopoietic progenitor cell (HPC). ③ Nude mice did not have neoplasia. CONCLUSION: Human cord blood CD34+ cells of transferring JAK2 genes may cooperate with other cytokines to amplify cord blood stem/progenitor cells in vitro for long. Therefore, this is potentially valuable for stem cells to treat some hereditary hematologic disease.

Objective To investigate the role of the mass screening by comparing the pathological features of prostate cancer between mass screening patients and clinical patients.Methods 107 cases of prostate cancer(including 51 patients from clinical diagnosis and 56 patients from mass screening) and 7 cases of prostate intraepithelial neoplasia(PIN,from mass screening) were analyzed using the Gleason’s grade system.Results ① Gleason’s grade of prostate cancer in mass screening group was lower than that in clinical diagnosis group(?2 =48.22,P

OBJECTIVETo study the membrane mobility of aquaporin 7 (AQP7) by cloning stably transfected CHO cells with expression of pEGFP-C1-AQP7, in which AQP7 cDNA was fused downstream and in frame to pEGFP-C1 gene.

METHODSThe full sequence of AQP7 was amplified by RT-PCR and then recombined in the downstream of the green fluorescent protein gene in the pEGFP-C1 vector. The recombinant vector pEGFP-C1-AQP7 was stably transfected into CHO cells. With fluorescent microscopy, immunocytochemical stain and Western blot, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.

RESULTS(1) The sequence of AQP7 cDNA of the Wistar rat was logged into the GenBank (access number: AY157737). (2) Identification demonstrated that pEGFP-C1-AQP7 fusion protein stably expressed in CHO cells. (3) With fluorescence microscopy, pEGFP-C1-AQP7 showed a predominant intracellular vesicular localization.

CONCLUSIONThe CHO cell line with stable pEGFP-C1-AQP7 expression was set up successfully for advanced research.

Animals ; Aquaporins ; biosynthesis ; genetics ; CHO Cells ; Cricetinae ; Cricetulus ; DNA, Complementary ; genetics ; Green Fluorescent Proteins ; biosynthesis ; genetics ; Male ; Molecular Sequence Data ; Rats ; Rats, Wistar ; Recombinant Fusion Proteins ; biosynthesis ; Reverse Transcriptase Polymerase Chain Reaction ; Testis ; metabolism ; Transfection

AIM:Aquaporins (AQP) are very important for the water transport across cell membrane. There are at least 10 mammalian AQPs( aquaporins 0-9) distributed in various organs and different kinds of cells. Each AQP has a distinct organ distribution, and this distribution could be useful in presuming the biological function of the aquaporin. The aim of this study was to figure out the distribution of aquaporin 7 (AQP7) and aquaporin 8(AQP8).METHODS：Semi-quantified RT-PCR was employed in this research. The ratio of OD value of target gene products divided by which of control gene products was calculated. Among 35 organs, testis, epididymis, skin, muscle, rectum, lung, bronchus, lymph node, stomach, duodenum, jejunum, ileum, colon, pancreas, liver, gall bladder, spleen, mammary gland, uterus, placenta, tonsil, urinary bladder, thyroid came from normal area of removed samples during operation. cDNA library of Prostate, thymus, salivary gland, penis, carotiol artery, adrenal gland, occipital lobe of brain, temporal lobe of brain, frontal lobe of brain, parietal lobe of brain, mid brain, choroid plexus are purchased from OriGene biotechnique company.RESULTS：①AQP 7 mRNA was found in testis, muscle, gall bladder, carotiol artery, lymph node and adrenal gland, and maximum expression of AQP 7 was in testis.②AQP 8 mRNA was found in pancreas, testis, skin and colon. and maximum expression of AQP 8 was in pancreas.CONCLUSION:Coexistence of AQP 7 and 8 in testis was confirmed, which suggested that both of these two aquaporins were involved in the regulation of testis function.

AIM: Aquaporins(AQP) are very important for the water transport across cell membrane. Aquaporin 7(AQP 7) and aquaporin 8(AQP8) expressed in the rat testis, but the biophysical functions of these two aquporins in testis and the regulatory mechanism of their expression remain unclear. The aim of this study was to find out if the expression of these two aquporins was correlative with the function of testis, and if panaxadiol saponins (PDS) affected their expression. METHODS: 4 weeks, 8 weeks and 12 weeks old rats were divided into saline and PDS injection groups. Semi-quantified RT-PCR method was employed to detect aquaporin 7 and 8 gene expression: using total RNA extracted from rat testis as PCR template, then using optical scanner to measure the OD value of bands on agrose electrophoresis. OD value of target gene products was divided by which of contol gene. The ratio was identified as the quantity of target gene expression. Western blot was also employed to detect expression of these two proteins in the testis. RESULTS: ①OD value of AQP 7 mRNA products in testis of 4 weeks, 8 weeks, 12 weeks rats were 48 227, 51 536, 59 567 respectively and the ratio divided by OD value of control gene was 0.82, 0.85, 0.99; OD value of AQP 8 mRNA products in testis of 4 weeks, 8 weeks, 12 weeks rats were 23 092, 39 302, 43 316 respectively, the ratio divided by OD value of control gene was 0.39, 0.65, 0.72. Thus, AQP 7 and 8 mRNA expression increased with age. ②After PDS injection for 2 weeks. AQP 7 and 8 mRNA expression in 4 weeks old rats increased to 0.97 and 0. 72,which in control group were 0. 82 and 0. 39; the ratio decreased to 0.77 and 0.55 in 8 weeks old rats, which in control group were 0.85 and 0.65. There was no PCR product of neither aquaporins in 12 weeks old PDS injected rats, except products of control gene. ③ Western blot of AQP 7 and 8 protein showed little difference between PDS and saline injected 12 weeks old rats. CONCLUSION: ①Quantity of AQP 7 and AQP 8 expression was related to testis function of rats. ②PDS influenced the expression of AQP 7 and AQP 8 mRNA, perhaps by promoting the secretion of LH in pituitary.