The clinical application of color Doppler ultrasound in guiding the puncture of new internal fistula in deep arteriovenous / 中国实用护理杂志

Objective:To investigate the clinical value of color Doppler ultrasound guidance in the puncture of new internal fistula in deep arteriovenous of hemodialysis patients.Methods:Selected thirty patients with arteriovenous fistula not less than 5 mm deep in the Department of Hemodialysis Room of Wuhan First Hospital, and they were divided into observation group and control group by the method of random number generator in IBM SPSS Statistics 22.0 software, with 15 patients in each group. The control group was treated with conventional blind puncture by experienced nurses (849 times), while the observation group was treated with color Doppler ultrasound guided puncture (855 times). By comparing two groups of deep arteriovenous fistula in new fistula patients within 6 months of the one-time success rate of puncture, the incidence of adverse events such as subcutaneous hematoma, bleeding, pain, interruption or postponement of treatment, and accidental injury to arteries caused by puncture errors of internal fistula, and the patient′s satisfaction with the puncture and dialysis results was evaluated to observe the advantage of color Doppler ultrasound guidance in clinical application of deep arteriovenous new fistula puncture.Results:The success rate of one-time puncture for internal fistula puncture in observation group was 98.95% (846/855), which was significantly higher than that in control group 81.27% (690/849), and the difference was statistically significant ( χ2 = 149.753, P < 0.01). The incidence of subcutaneous hematoma, bleeding, pain, interruption or delayed treatment in observation group respectively was 0.58% (5/855), 0.94% (8/855), 0.47% (4/855) and 0, however, in control group was 17.20% (146/849), 2.47% (21/849), 2.12% (18/849) and 1.06% (9/849), respectively. There were significant differences between the two groups ( χ2=145.557, 6.022, 7.875, Fisher′s exact test, P < 0.01 or 0.05). The incidence of arterial injury was extremely low in clinical practice, with 0 in observation group and 0.12 %(1/849) in control group, and the difference was not statistically significant ( P>0.05). The total incidence of adverse events caused by internal fistula puncture errors in the two groups was 1.99% (17/855) and 22.97% (195/849), respectively, and the difference was statistically significant ( χ2=172.126, P < 0.01). The average rank of satisfaction in observation group was 1 261.25, which was higher than control group 440.86, and the difference was statistically significant ( Z=-38.107, P < 0.01). Conclusions:Color Doppler ultrasound guidance in the puncture of new internal fistula in deep arteriovenous of hemodialysis patients has important clinical application value. It can significantly improve the success rate of one-time new internal fistula puncture, reduce the occurrence of adverse events due to puncture failure, and reduce patient′s pain, which is worthy of clinical promotion.

The clinical application of color Doppler ultrasound in guiding the puncture of new internal fistula in deep arteriovenous / 中国实用护理杂志

Objective@#To investigate the clinical value of color Doppler ultrasound guidance in the puncture of new internal fistula in deep arteriovenous of hemodialysis patients.@*Methods@#Selected thirty patients with arteriovenous fistula not less than 5 mm deep in the Department of Hemodialysis Room of Wuhan First Hospital, and they were divided into observation group and control group by the method of random number generator in IBM SPSS Statistics 22.0 software, with 15 patients in each group. The control group was treated with conventional blind puncture by experienced nurses (849 times), while the observation group was treated with color Doppler ultrasound guided puncture (855 times). By comparing two groups of deep arteriovenous fistula in new fistula patients within 6 months of the one-time success rate of puncture, the incidence of adverse events such as subcutaneous hematoma, bleeding, pain, interruption or postponement of treatment, and accidental injury to arteries caused by puncture errors of internal fistula, and the patient′s satisfaction with the puncture and dialysis results was evaluated to observe the advantage of color Doppler ultrasound guidance in clinical application of deep arteriovenous new fistula puncture.@*Results@#The success rate of one-time puncture for internal fistula puncture in observation group was 98.95% (846/855), which was significantly higher than that in control group 81.27% (690/849), and the difference was statistically significant (χ2 = 149.753, P < 0.01). The incidence of subcutaneous hematoma, bleeding, pain, interruption or delayed treatment in observation group respectively was 0.58% (5/855), 0.94% (8/855), 0.47% (4/855) and 0, however, in control group was 17.20% (146/849), 2.47% (21/849), 2.12% (18/849) and 1.06% (9/849), respectively. There were significant differences between the two groups (χ2=145.557, 6.022, 7.875, Fisher′s exact test, P < 0.01 or 0.05). The incidence of arterial injury was extremely low in clinical practice, with 0 in observation group and 0.12 %(1/849) in control group, and the difference was not statistically significant (P>0.05). The total incidence of adverse events caused by internal fistula puncture errors in the two groups was 1.99% (17/855) and 22.97% (195/849), respectively, and the difference was statistically significant (χ2=172.126, P < 0.01). The average rank of satisfaction in observation group was 1 261.25, which was higher than control group 440.86, and the difference was statistically significant (Z=-38.107, P < 0.01).@*Conclusions@#Color Doppler ultrasound guidance in the puncture of new internal fistula in deep arteriovenous of hemodialysis patients has important clinical application value. It can significantly improve the success rate of one-time new internal fistula puncture, reduce the occurrence of adverse events due to puncture failure, and reduce patient′s pain, which is worthy of clinical promotion.

Application of retrobulbar nerve block combined with general anesthesia in corneal transplantation / 器官移植

Objective To explore the application value of retrobulbar nerve block combined with general anesthesia in the penetrating keratoplasty (PKP). Methods A total of 100 recipients undergoing PKP from January 2017 to January 2019 were recruited in this study. All recipients were divided into the observation group (n=50) and control group (n=50) by random number table method. In the control group, patients received laryngeal mask airway under general anesthesia, and in the observation group, patients received laryngeal mask airway under general anesthesia combined with retrobulbar nerve block. Hemodynamic changes of the PKP recipients before and after operation were observed in two groups. The dosage of analgesic drugs and the incidence of complications were observed in two groups. The degree of pain at postoperative 2-, 6- and 24-h was evaluated by visual analogue scale (VAS) in two groups. The awakening situation of the recipients in two groups was observed. The levels of inflammatory cytokines at 1 d before and after operation were statistically compared in two groups. Results The average arterial pressure and heart rate at intraoperative 15 min and after the surgery in the observation group were significantly higher than those in the control group (both P < 0.05). In the observation group, the dosage of remifentanil and propofol were (1.0±0.4) mg and (299±40) mg, significantly lower than (1.3±0.6) mg and (365±42) mg in the control group (both P < 0.05). The incidence of complications did not significantly differ between two groups (P > 0.05). In the observation group, the VAS scores at 2-, 6- and 12-h after operation were remarkably lower than those in the control group (all P < 0.01). The respiratory recovery time, eye opening time, directional force recovery time and extubation time of the recipients in the observation group were significantly shorter than those in the control group (all P < 0.05). The expression levels of including interleukin (IL)-1, IL-6 and tumor necrosis factor-α (TNF-α) at postoperative 1 d in the observation group were considerably lower than those in the control group (all P < 0.05). Conclusions Retrobulbar nerve block combined with general anesthesia can maintain hemodynamic stability during PKP, reduce the dosage of remifentanil and propofol and alleviate the degree of postoperative pain and inflammatory responses of the recipients.

LIM kinases and their roles in the nervous system / 浙江大学学报·医学版

LIM kinase-1 (LIMK1) and LIM kinase-2 (LIMK2) are kinases that have serine/threonine and tyrosine dual specificity. Although they show significant structural similarity, LIMK1 and LIMK2 have different expression patterns, subcellular localization, and functions. Activation of LIM kinases regulates the downstream of Rho GTPases, and influences the architecture of the actin cytoskeleton by regulating the activity of cofilin. Recent studies have shown that LIM kinases play important roles in the nervous system. For example, development of the central nervous system is reliant upon the presence of LIMK1, and deletion of Limk1 gene is involved in the development of the human genetic disorder Williams syndrome. Therefore, it is of vital physiological significance to investigate the neuronal function of LIM kinases. In this review, we outline the structure, phosphorylation regulation and neuronal function of LIM kinases, so as to provide new ideas for the treatment of these neurological diseases.

Clinical observation of corneal lamellar debridement combined with sutureless amniotic membrane transplantation for the treatment of superficial fungal keratitis / 国际眼科杂志(Guoji Yanke Zazhi)

To evaluate the clinical efficacy of corneal lamellar debridement combined with sutureless amniotic membrane transplantation for the treatment of superficial fungal keratitis. ●METHODS:Totally 22 cases (22 eyes) with superficial fungal keratitis were referred to our hospital from April 2012 to October 2013. The patients with persistent cornea ulcer after treatment of local and systemic antifungal drugs underwent corneal lamellar debridement combined with sutureless amniotic membrane transplantation, and the recipient bed was covered with an amniotic membrane using fibrin sealant during the operation. All patients were still given topical antifungal therapy for 1 -2mo after operation. The followed- up time was 3mo or above. We observed the corneal healing and amniotic membrane adhesion by split lamp microscope, and investigated the transformation of amniotic membrane and fungal infection recurrence with confocal microscope. ●RESULTS: Corneal edema and anterior chamber reaction of 21 patients disappeared gradually, and no amniotic membrane graft dissolved and shed off within 1-2wk postoperatively. Two weeks after operation, the graft integrated into the corneal and the corneal wounds' thickness increased gradually, the corneal epithelium reconstructed and corneas became clear. Four weeks after operation, the corneal scarring developed gradually and fluorescence staining was negative. Nineteen cases' amniotic membranes that adhered with the cornea dissolved 4wk after operation. There were different degrees of corneal nebula or macula remained 3mo postoperatively. All patients' vision improved in varying degrees, except in 1 case with fungal keratitis who had been cured by lamellar keratoplasty. ●CONCLUSlON:Corneal lamellar debridement combined with sutureless amniotic membrane transplantation can effectively remove the foci of inflammation, improve the local efficacy, shorten the operation time, relieve the postoperative reaction, and promote cornea union, which is an effective way to treat superficial fungal keratitis.

An investigation of iodine nutrition and thyroid function in downtown residents of Longyan city, Fujian province / 中国地方病学杂志

Objective To find out the iodine nutritional status of Longyan downtown residents,evaluate the effectiveness of control measures and provide a scientific basis for developing control strategies.Methods Infants aged 0 to 2 year-old,children aged 8 to 10,adults aged 18 to 45 and pregnant and lactating women were selected as survey subjects.Children goiter was detected with B ultrasound.Residents per capita daily salt intake was investigated by weighing method.Three urinary samples and a milk sample of lactating women were randomly collected.Urinary iodine and milk iodine content were determined by arsenic cerium catalytic spectrophotometric assay.Blood samples were collected and thyroid function (including serum TT3,FT3,TT4 and FT4) and thyroid stimulating hormone(TSH) were measured with direct chemiluminescence immunoassay,and thyroglobulin antibody(TgAb),thyromicrosome antibody(TMAb),and thyroglobulin (Tg) were measured with radioimmunoassay (RIA) in serum.ResultsThe goiter rate of children aged 8 to 10 was 1.8％ (2/110),and median thyroid volume was 2.75 ml.Household iodized salt coverage rate was 100.00％(318/318),and qualified iodized salt was 94.03％ (299/318).The daily per capita salt intake was (6.13 ± 3.56)g.The average medians of urinary iodine of the infants,children,adults,pregnant and lactating women were 181.8,315.2,196.6,158.7,136.4 μg/L,respectively.The median of milk iodine of lactating women was 155.6 μg/L.The proportions of serum TT3,FT3,TT4,FT4 and TSH which higher than normal were 3.6％ (11/308),0.6％ (2/309),23％ (7/309),1.0％ (3/313) and 1.3％ (4/312),respectively.While the proportions of serum TT3,FT3,TT4,FT4 and TSH that lower than normal were 2.3％ (7/308),11.7％(36/309),2.3％(7/309),12.8％(40/313),and 1.6％(5/312),respectively,of which 16 cases of both TgAb and TMAb were higher than normal.ConclusionsExisting salt iodine level is appropriate for 0 to 2 year-old infants and young children,18 to 45 year-old adults,pregnant and lactating women in downtown Longyan city.The iodine intake of children aged 8 to 10 is excessive.Thyroid function monitoring is recommended to be included in the routine monitoring.

High-Definition Computed Tomography for Coronary Artery Stent Imaging: a Phantom Study

OBJECTIVE: To assess the performance of a high-definition CT (HDCT) for imaging small caliber coronary stents (< or = 3 mm) by comparing different scan modes of a conventional 64-row standard-definition CT (SDCT). MATERIALS AND METHODS: A cardiac phantom with twelve stents (2.5 mm and 3.0 mm in diameter) was scanned by HDCT and SDCT. The scan modes were retrospective electrocardiography (ECG)-gated helical and prospective ECG-triggered axial with tube voltages of 120 kVp and 100 kVp, respectively. The inner stent diameters (ISD) and the in-stent attenuation value (AVin-stent) and the in-vessel extra-stent attenuation value (AVin-vessel) were measured by two observers. The artificial lumen narrowing (ALN = [ISD - ISDmeasured]/ISD) and artificial attenuation increase between in-stent and in-vessel (AAI = AVin-stent - AVin-vessel) were calculated. All data was analyzed by intraclass correlation and ANOVA-test. RESULTS: The correlation coefficient of ISD, AVin-vessel and AVin-stent between the two observers was good. The ALNs of HDCT were statistically lower than that of SDCT (30 +/- 5.7% versus 35 +/- 5.4%, p < 0.05). HDCT had statistically lower AAI values than SDCT (15.7 +/- 81.4 HU versus 71.4 +/- 90.5 HU, p < 0.05). The prospective axial dataset demonstrated smaller ALN than the retrospective helical dataset on both HDCT and SDCT (p < 0.05). Additionally, there were no differences in ALN between the 120 kVp and 100 kVp tube voltages on HDCT (p = 0.05). CONCLUSION: High-definition CT helps improve measurement accuracy for imaging coronary stents compared to SDCT. HDCT with 100 kVp and the prospective ECG-triggered axial technique, with a lower radiation dose than 120 kVp application, may be advantageous in evaluating coronary stents with smaller calibers (< or = 3 mm).

Expression of high mobility group box chromosomal protein 1 in mice with lupus nephritis / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To determine the role of the novel proinflammatory cytokine high mobility group box chromosomal protein 1 (HMGB-1) in the pathogenesis of lupus nephritis.</p><p><b>METHODS</b>Serum levels of anti-dsDNA antibodies were determined by enzyme linked immunosorbent assay (ELISA). Renal morphologic features were examined by light microscopy, electron microscopy, and immunohistologic analyses. The mRNA expression of HMGB-1 and monocyte chemoattractant protein-1 (MCP-1) was detected by RT-PCR.</p><p><b>RESULT</b>MRL/lpr mice demonstrated characteristic alterations of serum immune parameters, with progressively increased anti-dsDNA antibodies with age, compared with age-matched C57BL/6J mice. MRL/lpr mice showed progressive development of renal damage, starting at 12 weeks of age and reached the peak at 20 weeks. The observed lesions included the presence of enlarged hypercellular glomeruli, with increased numbers of both resident cells and infiltrating leukocytes. Higher expression of HMGB-1 mRNA was found in MRL/lpr mice than what in C57BL/6J mice. Expression of HMGB-1 was positively correlated with that of MCP-1 mRNA.</p><p><b>CONCLUSION</b>The results demonstrate that the higher expression of HMGB-1 may contribute to the pathogenesis of lupus nephritis.</p>

The relationship between infection of helicobacter pylori and iron deficiency anemia / 中国基层医药

Objective To investigate the relationship between infection of helicobaeter pylori(Hp) and iron deficiency anemia(IDA) and to explore effective clinical treatments for patients with Hp associated IDA . Method 1. The Hp infection ratio of 40 chronic gastritis with IDA and 42 patients without IDA were counted up respectively. 2. 36 patients with Hp-positive chronic gastritis were divided into two groups randomly. One group was treated by using Hp eradication therapy in conjunction with oral iron supplement and the other using iron supplement only. The hema-tological parameters before and after treatment are measured and the effectiveness of the treatment was evaluated. Re-sult The Hp infection ratio in chronic gastritis patients with IDA is |figher than that in patients without IDA and the difference is significant. After Hp eradication therapy in conjunction with iron supplement Hb. serum iron and serum ferritin level in Hp associated IDA patients are increased significantly while that for patients treated by using iron sup-plement only have no significant improvement. Conclusion It appears that Hp infection may be related to IDA. When iron supplement treatment has no obvious effect to an IDA patient,lt may suggest a ease of Hp associated IDA. The i-ton supplement treatment has positive effects to IDA patients after Hp eradication.

Expression of B7-H4 in prostate cancer and its clinical significance / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the expression of B7-H4 in prostate cancer tissue and the relationship between the expression and the clinicopathological features.</p><p><b>METHODS</b>Immunohistochemical staining was used to detect the expression of B7-H4 in prostate cancer tissue. And the relationship between the expressions and pathology was evaluated.</p><p><b>RESULTS</b>The B7-H4 was diffusely expressed in cytoplasm and/or membrane of the prostate cancer tissue; the expression was much higher than that in normal prostate tissue (P<0.05). The expression of B7-H4 in the prostate cancer tissue was higher in patients with higher tumor grade.</p><p><b>CONCLUSION</b>B7-H4 may be used as an new indicator for the diagnosis and prognosis of prostate cancer and a novel target for immunotherapy.</p>

Effects of Sema3A derived from tumor cells on functions of dendritic cells / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effects of tumor cell-derived Sema3A on the immunological functions of murine dendritic cells (DCs).</p><p><b>METHODS</b>Lung adenocarcinoma A549 cells were transfected with small interference RNA, Si-Sema and Si-mut, and the interference efficiency was determined by real-time PCR and Western-blot. The concentrated supernatants from cultured tumor cells, Si-Sema and Si-mut-infected tumor cells were subjected to DCs respectively. The immunophenotypes of DCs were analyzed by flow cytometry, the production of IL-12P70 and the ability of DCs to stimulate DO11. 10 T cells secreting IFN-gamma and IL-2 were detected by enzyme linked immunosorbent assay (ELISA).</p><p><b>RESULTS</b>Knockdown with Si-Sema3A significantly decreased the secretion of Sema3A by A549 cells in comparison with the Si-mut cells. DCs exposed to supernatants from Si-Sema cells showed elevated levels of MHC, CD40 and CD80, more production of IL-12P70, and enhanced capability of activating antigen-specific T cells, as evidenced by the remarkably increased levels of IFN-gamma and IL-2.</p><p><b>CONCLUSION</b>A549 cells secrete Sema3A to inhibit the maturation and functions of DCs, which might be associated with the unidentified mechanism of immune evasion by tumor cells.</p>

Expression and identification of recombinant mouse gene B7-H4 / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct a eukaryotic expression vector encoding the gene of extracellular region of mouse B7-H4, to express it in yeast cell line and to determine its biological activity.</p><p><b>METHODS</b>The extracellular region of the mouse B7-H4 gene was amplified with Xho I and EcoR I by PCR from a mouse B7-H4 chimeric plasmid. Digested with Xho I and EcoR I, the mB7-H4 gene was inserted into the yeast expression plasmid Ppic9. The DNA sequence was confirmed by double digestion and the Ppic9-mB7-H4 plasmid was transfected into the yeast cells. The expression of mB7-H4 was confirmed by PCR, Western Blot and ELISA analysis, and its biological function was determined.</p><p><b>RESULT</b>Ppic9-mB7-H4 transfectants expressed mB7-H4 in yeast cells, and mB7-H4 effectively inhibited the proliferation of T lymphocytes with a fractional inhibition rate of 28.3 % and inhibited IL-2, IL-4, IL-10 and IFN-gamma production with fractional inhibition rates of 68.8%, 78.8%, 67.6% and 77.7%, respectively.</p><p><b>CONCLUSION</b>The eukaryotic expression plasmid mouse B7-H4 has been successfully constructed and the expressed products of B7-H4 possess biological activity.</p>

Construction and identification of recombinant adenoviral vector encoding B7-H4 gene / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct the recombinant adenovirus containing B7-H4 gene with AdEasy XL system and to identify its biological activities.</p><p><b>METHODS</b>The full-length mouse B7-H4 gene was amplified by RT-PCR from C57 mouse lung and put into T vector, then verified by sequencing. Digested with Xhol I and EcoR V the B7-H4 gene was inserted into pshuttle-CMW(PSC). Pme I linearized shuttle plasmid was transformed into E.coli BJ5183-AD-1 to obtain the recombinant adenoviral plasmid pAd-mB7-H4 by efficient homologous recombination. Then the recombinant adenovirus-mB7-H4/Ad was obtained by packaging Pac I linearized in D-293 cells. The mRNA and protein expression of B7-H4 in mB7-H4/Ad infected AD-293 cells were detected by RT-PCR and Western blot, respectively. mB7-H4/Ad was used to infect L929 cells, the bioactivity of expressed B7-H4 in stimulation of T lymphocytes proliferation and cytokine production were tested.</p><p><b>RESULTS</b>The full-length of mB7-H4 was cloned from mouse lung tissue cDNA and verified by sequencing. The recombinant plasmid pAd-m B7-H4 was successfully generated by homologous recombination, and the primary mB7-H4/Ad was obtained by packaging pAd-B7-H4 in AD-293 cells. Compared with the negative control, L929 cells infected with mB7-H4/Ad effectively inhibited the proliferation of T lymphocytes and cytokines production.</p><p><b>CONCLUSION</b>The bioactive recombinant adenovirus mB7-H4/Ad has been successfully constructed.</p>

Diagnosis and treatment of pancreatic trauma / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To improve diagnosis and therapeutic efficacy for pancreatic trauma.</p><p><b>METHODS</b>We retrospectively analyzed 71 cases of pancreatic injuries received in our department from 1987 to 2004. Different surgical procedures were performed according to different patterns of pancreatic injuries. Among them, 31 cases were defined as Grade I or II injury according to the pancreatic organ injury score developed by American Association for the Surgery of Trauma and were performed pancreas debridement and drainage; 26 cases belonged to Grade III injury and were performed distal pancreatectomy plus external drainage; 10 cases of Grade IV injury in whom 8 were performed distal Roux-en-Y pancreaticojejunostomy and 2 were performed Whipple operation; 4 cases of Grade V injury in whom 1 was performed restoration of duodenal damage, suture of proximal pancreatic laceration and distal Roux-en-Y pancreaticojejunostomy, and 3 were performed Whipple operation.</p><p><b>RESULTS</b>Sixty-six cases were cured, of whom 14 had postoperative complications, and 5 cases died. The causes of death were of pancreatic fistula in 2 cases, upper gastrointestinal bleeding in 1 case, ARDS in 1 case, and serious abdominal infection in 1 case.</p><p><b>CONCLUSIONS</b>Preoperative diagnosis for pancreatic trauma is rather challenging. Prompt explorative laparotomy is still a reliable diagnostic method for pancreatic trauma. In order to improve curative rate, different surgical procedures should be undertaken according to different sites and grades of pancreatic traumas.</p>

Construction of the eukaryotic expression vector containing rat transforming growth factor beta type II receptor and interferon gamma fusion genes / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To express a bioactive fusion protein comprising rat soluble transforming growth factor beta type II receptor and interferon gamma(rsTGFbetaR II-IFN gamma) in mammalian cells.</p><p><b>METHODS</b>mRNA was extracted from rat liver and the sTGFbetaR II-IFN gamma genes amplified by RT-PCR, then the two gene segments were cloned into the same pSecTag2A expression vector, and pSecTag2A/rsTGFbetaR II-IFN gamma recombinant plasmid was obtained, which was later transfected into CHO cells using liposomes. The expression of pSecTag2A/rsTGFbetaR II-IFN gamma in the supernatant was detected by ELISA and Western blotting. The bioactivities of the fusion protein were tested by sTGF betaR II-IFN gamma and IFN gamma bioassays.</p><p><b>RESULTS</b>pSecTag2A/rsTGFbetaR II-IFN gamma transfectants expressed rsTGF betaR II-IFN gamma fusion protein. The purified fusion protein exhibited anti-viral activity and antagonized the proliferation-inhibitive effect of TGFbeta1 on CCL-64 cells. It inhibits the HSC activation in vitro.</p><p><b>CONCLUSION</b>The pSecTag2A/rsTGFbetaR II-IFN gamma recombinant plasmid constructed in this study can express bioactive rsTGFbetaR II-IFN gamma fusion protein.</p>

Construction and expression of the eukaryotic expression vector containing human soluble transforming growth factor beta receptor II / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To construct a eukaryotic expression vector encoding the gene of extracellular region of type II transforming growth factor beta receptor (sTGFbetaR II), to express the protein in CHO cell line and to determine its biological activity.</p><p><b>METHODS</b>The extracellular region (amino acids 1-159) of the human TGFbetaR II cDNA was amplified by PCR from a TGFbetaR II chimeric plasmid,and the eukaryotic expression plasmid pCDNA3.1/myc-his(-)B-sTGFbetaR II(pCDNA-sTGFbetaR II) was constructed by inserting the sTGFbetaR II cDNA into the EcoR I/Hind III-digested pCDNA. The DNA sequence was confirmed by double digestion and the pCDNA-sTGFbetaR II plasmid was transfected into the CHO cell line. The sTGFbetaR II protein was confirmed by Western blotting analysis and its biological function was determined.</p><p><b>RESULTS</b>The specific protein was observed in western blotting, and the protein abrogated the growth-inhibitory effects of TGF-beta1 on mink lung epithelial cells (Mv1Lu).</p><p><b>CONCLUSION</b>The eukaryotic expression plasmid pCDNA-sTGFbetaR II has been successfully constructed and the sTGFsTGFbetaR II protein with biological activity obtained.</p>

Effect of Triptolide on TNFalpha-induced activation of NF-kappaB and expression of COX-2 and iNOS in human rheumatoid arthritis synovial fibroblasts / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To explore the effects of Triptolide (TP) on TNFalpha-induced cell proliferation and expressions of cyclooxygenase-2 (COX-2), inducible nitric oxide synthase (iNOS) and their inducing products PGE2, NO in human rheumatoid arthritis synovial fibroblasts (RASF).</p><p><b>METHODS</b>Fibroblasts (RASF) were obtained from synovial tissue of patients with RA and were cultured in vitro. RASF were stimulated with TNFalpha(20 microg/L) in the presence or absence of TP(0 - 100 microg/L) for 20 h. The RASF proliferation was determined by (3)H-TdR incorporation, and the productions of PGE2 and NO in culture supernatants of RASF were detected with competitive ELISA and enzyme reduction of nitrate. Expressions of COX-2 and iNOS mRNA in RASF were analyzed by semi-quantitative RT-PCR. Expressions of COX-2 and iNOS protein were estimated by Western-blot method and cellular enzyme immunoassay in synovial fibroblasts. NF-kappaB activity in whole-cell extract of RASF was also measured by an ELISA-based method.</p><p><b>RESULTS</b>TP (>20 microg/L) down-regulated markedly TNFalpha-induced COX-2 and iNOS mRNA and protein expression, and their inducing products PGE2 and NO of synovial fibroblasts. This effect was positively correlated with TP concentrations. NF-kappaB activity in TNFalpha-stimulated synovial cells was suppressed profoundly by TP treatment (IC(50) approximately 35microg/L). The activity of NF-kappaB was correlated with the levels of COX-2 and iNOS expression in TNFalpha-stimulated RASF. No change was observed in proliferation of synovial cells after treatment of TP.</p><p><b>CONCLUSION</b>TP could significantly down-regulate TNFalpha-induced COX-2, iNOS expression and production of PGE2, NO in human RASF, which is associated with the suppression of NF-kappaB activity.</p>

Effects of intrasplenic transplantation of IL-18 gene-modified fetal hepatocytes on mouse immune function / 浙江大学学报·医学版

OBJECTIVE: To investigate the effects of IL-18 gene-modified fetal hepatocytes (AdmIL-18/MNL.CL2) intrasplenic transplantation on mouse immune function. METHODS: Forty mice were evenly divided into 4 groups of 10. Each group received an intrasplenic transplantation one of the following: AdmIL-18/BNL.CL2, Ad-LacZ/BNL.CL2 (virus control), BNL.CL2 (cell control) and PBS (blank control). After two weeks, the mice were sacrificed. Serum cytokine levels, Mpsi and splenic cell culture supernatant and liver tissue extracts supernatants were measured using ELISA. Hepatic cytokines mRNA expression were determined by RT-PCR. THe cytotoxicity of peritoneal Mpsi and NK activity of spienocytes were detected by LDH release assay. The proliferation of splenic lymphocytes was determined by MTT assay. RESULTS: The IL-18, IL-2,IFN-gamma, TNF-alpha levels of serum, Mpsi and splenocyte culture supernatant, liver tissue extracts supernatants in mice transplanted with AdmIL-18/BNL.CL2 were higher and the IL-4, IL-10 levels were lower compared to their levels in other 3 groups. The highest IL-18, IL-2, IFN-gamma, TNF-alpha and the lowest IL-4, IL-10 mRNA expressions in the liver were observed in mice transplanted with AdmIL-18/BNL.CL2. The mice transplanted with AdmIL-18/BNL.Cl2 showed significantly increases cytotoxicity of Mpsi, NK activity and splenic cell proliferation compared with the other 3 groups. CONCLUSION: AdmIL-18 can be effectively transfected into mice fetal heptocytes which subsequently IL-18. Intransplenic transplantation of IL-18 gene-modified fetal hepatocytes may augment mouse immune function and provide an useful basis for targeted gene therapy of liver disease.

Serum levels of sFas, sICAM-1, IL-18 in patients with chronic hepatitis C and their clinical significance / 浙江大学学报·医学版

OBJECTIVE: To investigate the serum levels of soluble Fas antigen (sFas), soluble intercellular adhesion molecules-1 (sICAM-1), interleukin-18 (IL-18) in patients with chronic hepatitis C and to study their roles in pathogenesis of chronic hepatitis C. METHODS: Serum sFas, sICAM-1, IL-18 levels were measured in 30 cases of chronic hepatitis C before and after treatment of interferon-alpha by enzyme-linked immunosorbent assay (ELISA), serum titer of HCV-RNA was detected by quantitative PCR and serum ALT activity was also detected. RESULTS: Serum levels of sFas sICAM-1 IL-18 in chronic hepatitis C patients were significantly higher than those in normal controls (P<0.01), showing correlation with serum HCV-RNA titer (r=0.915, r=0.795, r=0.757, respectively, P<0.01), Serum levels of sICAM-1, IL-18 showed correlation with serum ALT level(gamma=0.952, gamma=0.969, respectively, P<0.01), but no relationship was observed between serum sFas and serum ALT level(P>0.05). Serum levels of sFsa sICAM-1 IL-18 markedly decreased in responsive patients while no change was observed in patients with no response after treatment. CONCLUSION: Soluble Fas, soluble ICAM-1, IL-18 may participate in the pathogenesis of chronic hepatitis C and show correlation with the severity of histological inflammation and viral titer.