Objective To evaluate the efficacy,adverse events of gemcitabine vs.5-FU with radiotherapy for locally advanced pancreatic carcinomas.Methods Between January 2007 and January 2011,a total of 56 patients with locally advanced pancreatic carcinomas was included and clinical data were retrospectively analyzed.All patients received 3-DCRT radiotherapy with individual dose of 1.8 ～ 2 Gy,5 times per week,and total dose of 45 ～ 50.4 Gy for 25 ～ 28 fractions,and received concurrent chemotherapy (5-FU or gemcitabine).The patients (n =30) in gemcitabine group were treated with gemcitabine (500 rng/m2 at the 1st,8th,15th,22nd day,at 10 mg · (m2)-1 · min-1,through micro-pump) during radiotherapy; 3 weeks after radiotherapy the patients received gemcitabine infusion at a dose of 800 mg · (m2)-1 · d-1,one time per week,for 3 or 4 cycles.The patients (n =26) in 5-FU group were treated with 5-FU (500 mg/m2 at the 1 ～ 5th day per week,IV),the cycle was repeated every 2 weeks during radiotherapy; 3 weeks later the patients received 5-FU infusion at a dose of 800 mg · (m2)-1 · d-1,the 1st ～5th day per week,the cycle was repeated every 2 weeks ; with a total of 3 or 4 cycles.The efficacy and adverse events were observed,and the patients were followed until June 2013,and the median survival,1 year and 2 year survival was calculated.Results Of the 56 patients,the overall response (CR + PR) rate was 73.2％,and it was 65.3％ in radiotherapy with 5-FU group,80.0％ in radiotherapy with gemcitabine group (P ＜ 0.05).The overall one and two year survival rate was 48.2％ and 14.3％,while median survival was 15.2 months,and the corresponding values were 42.3％,11.5％,13.3 months in radiotherapy with 5-FU group,and 53.3％,16.7％,16.6 months in radiotherapy with gemcitabine group,and the survival difference between the two groups was not statistically significant (P =0.071).At the end of treatment,the pain-relief rate (VAS score ＜4) of the 56 patients was 83.3％,it was 75.0％ in 5-FU group and 90.0％ in gemcitabine group.In radiotherapy with gemcitabine group,the incidence of 3～ 4 grade myelosuppression was significantly higher than that in radiotherapy with 5-FU group,and the difference between the two groups was statistically significant (20.0％ vs 7.6％,P ＜ 0.05).Conclusions For the locally advanced pancreatic carcinomas,radiotherapy with gemcitabine can improve pain-relief and prolong survival compared with radiotherapy with 5-FU,but the incidence of adverse event of myelosuppression is higher.

OBJECTIVETo explore the relationship between maternal milk and serum thyroid hormones in patients with thyroid-related diseases.

METHODSSerum and breast milk samples were collected from 56 breastfeeding mothers. Milk and serum free triiodothyronine (FT3), free thyroxine (FT4), triiodothyronine(T3), thyroxine (T4), and thyrotrophin (TSH) were determined, and T3/T4 was calculated. Using the serum thyroid hormones as the independent variables and milk thyroid hormones as the dependent variables, we performed linear regression analysis.

RESULTSThe milk FT3, FT4, T3, T4, TSH, and T3/T4 were (2.30 ± 0.82) pg/ml ,(0.45 ± 0.26) ng/dl, (0.35 ± 0.20) ng/ml, (2.96 ± 1.55) Μg/dl, (0.12 ± 0.08) ΜU/ml, and 0.12 ± 0.04, respectively. Milk FT3 (r = 0.778, P = 0.000), T3 (r = 0.603, P = 0.000), T4 (r = 0.485, P = 0.004), and TSH (r = 0.605, P = 0.000) concentrations were positively correlated with those in serum.

CONCLUSIONThyroid hormones are present in human milk and are positively correlated with those in serum.

Adult ; Female ; Humans ; Milk, Human ; chemistry ; Thyroid Diseases ; blood ; Thyroid Hormones ; blood ; chemistry ; Thyrotropin ; blood ; chemistry ; Triiodothyronine ; blood ; chemistry

Objective To explore the relationship of alcoholism between osteoporosis or femoral head necrosis.Methods In this case-control study,we selected 95 eligible patients with femoral head necrosis and another 67 cases of osteoporosis as case group,together with 342 patients of fractures from the Second Hospital affiliated to Shanxi Medical College,from February to December 2010,as the control group.Questionnaire was used to collect general information of the patients.Through comparative analysis,related factors of femoral head,osteoporosis were defined.18 patients with alcoholic femoral head necrosis,11 patients with alcoholic osteoporosis and 20 patients with fractures were selected from the above said three groups and going through the Alcohol Use Disorders Identification Test (AUDIT) as well as the Alcohol Use Disorders Scale (ADS).Using SPSS 13.0 conducted one-way ANOVA (analysis of variance),chi-square test,categorical logistic regression analysis were used for statistical analysis.Results Results from logistic regression analysis showed that the adjusted odds ratio of those subjects who liked drinking alcohol had an incidence of femoral head necrosis or osteoporosis as 7.70 (95％ CI:1.84,32.30) and 8.44 (95％ CI:1.70,41.90),respectively.The risks of using hormone for treating femoral head necrosis or osteoporosis were 78.43 (95％CI:11.20,149.05) and 22.75 (95％CI:2.59,100.27) times than those without.Data from the AUDIT showed that:over-dose of alcohol drinking habit existed 100％ in the femoral head necrosis group while 54.45％ in the osteoporosis group,while 75 percent patients in the fractures group had normal alcohol drinking habit.Statistically significant differences appeared in the three groups (P＜0.01).Results from the ADS showed that there were statistically significant differences between the ADS scores of the three groups (F=3.68,P=0.03).Conclusion Alcohol intake did seem to be highly correlated with the incidence rates of femoral head necrosis or osteoporosis.Alcohol-related necrosis could be viewed as alcohol-dependent diseases while alcohol-related and osteoporosis could partially be recognized as alcohol-dependent disease.

OBJECTIVEto investigate the effect of somatostatin on inflammatory immune disorders and prognosis in patients with severe sepsis caused by abdominal diseases.

METHODSfifty-three patients with severe abdominal sepsis (age > 18 years, APACHE-II score > 15) from June 2005 to June 2009 were randomly divided into Somatostatin group (n = 23) and SSC Group (n = 30). Fifteen healthy volunteers of the same age range were chosen as Control group. The SSC group was treated with classical SSC therapy, and the Somatostatin Group was treated with the same regime plus 14-peptide somatostatin continuous infusion at the dose of 6 mg/24 h for 7 days. The serum levels of interleukin-10 (IL-10), tumor necrosis factor-α (TNF-α) were determined by using ELISA. CD(4)(+), CD(8)(+) T cell subsets were determined by fluorescence activated cell sorter(FACS) and CD(4)(+)/CD(8)(+) was calculated. APACHE-II score was observed on admission (d1) and day 3, 7 and 14 after treatment. Morality rates in 28 days in two groups were recorded.

RESULTScompared with Control group, IL-10 and TNF-α levels were significantly elevated in patients with severe abdominal sepsis (P < 0.05), while CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) decreased significantly (P < 0.05). Compared with the Somatostatin group CD(4)(+), CD(8)(+) T cell and CD(4)(+)/CD(8)(+) on d7 and d14 in SSC Group were significantly increased (P < 0.05), while IL-10 and TNF-α decreased significantly(P < 0.05). APACHE-II scores on d3, d7, d14 of Somatostatin group were significantly lower than those of SSC group, and 28 d mortality rate also declined.

CONCLUSIONSin patients with severe abdominal sepsis, systemic inflammatory response and immune suppression exist simultaneously. Somatostatin has a dual immunomodulatory activity in these patients.

APACHE ; Case-Control Studies ; Female ; Humans ; Interleukin-10 ; blood ; Male ; Prognosis ; Prospective Studies ; Sepsis ; drug therapy ; etiology ; immunology ; Somatostatin ; therapeutic use ; T-Lymphocyte Subsets ; immunology ; Tumor Necrosis Factor-alpha ; blood

The purpose of this study was to construct 4 types of nonsense-mutated eukaryotic expression plasmids of fIX gene, using pcDNA3.1 plasmid containing fIX cDNA as template, and to identify, then to perform their expression in COS-7 cells. These stop mutants constructed by site-directed mutagenesis based on PCR, and further confirmed by DNA sequencing. COS-7 cells were transfected with either the wild-type or mutated fIX expression constructs, then the relative expression levels of fIX mRNA were detected by real time fluorescent quantitative PCR. The result showed that except the designed sites, there were no other nucleotide mutation in the sequences of four nonsense mutants. The results of real time PCR proved that the nonsense-mutated vectors can be effectively expressed in COS-7 cells. It is concluded that the nonsense-mutated eukaryotic expression vectors of fIX gene have been successfully constructed and can express in COS-7 cells, which provides the material basis for further researches on mechanism and treatment of FIX deficiency and the function defects caused by nonsense mutation.
Animals ; Base Sequence ; COS Cells ; Cercopithecus aethiops ; Cloning, Molecular ; Codon, Nonsense ; genetics ; Factor IX ; genetics ; Genetic Vectors ; Transfection

OBJECTIVETo explore the differences of PDGF and EGF expression in the wound healing between fatal and adult.

METHODSWith the established animal model of fetal scarless healing and the adult samples, an immunohistochemical technique was used to evaluate the expression of PDGF and EGF in the normal adult skin, normal fetal skin, and the process of their wound healing.

RESULTS1. The expression of the PDGF was not found in the fetal skin, but a mild amount of the PDGF was shown in the epidermis and the upper dermal layer 12 hours and 1 day after the wounding process. In the normal adult skin, expression of PDGF was shown in the dermal fibroblasts, macrophagocytes and blood capillaries, and a strong expression was presented during its wound healing process. 2. In the fetal skin, the expression of the EGF was seen in the epidermis, hair follicles, sebaceous glands and sweat glands, but there were no markedly changes during the wound healing. In the adult skin, a positive stain of the EGF was shown in the basal layer of the epidermis while the mild stain in hair follicles and sweat glands. The level of the expression became gradually decreasing with the time going in the wounded adult skin.

CONCLUSIONThe different expression of growth factors between fetal and adult skin in wound healing may be one of the important reasons that the fetal wound could produce scarless healing.

Adult ; Epidermal Growth Factor ; metabolism ; Epidermis ; metabolism ; Fetus ; Fibroblasts ; metabolism ; Hair Follicle ; metabolism ; Humans ; Platelet-Derived Growth Factor ; metabolism ; Skin ; injuries ; metabolism ; Sweat Glands ; metabolism ; Wound Healing

OBJECTIVETo investigate the effects of GnRH analogues GnRHa and GnRHant on the MAPK pathway in rat Leydig cells.

METHODSRat Leydig cells were primarily cultured for 24 hours in vitro and serum-starved for 2 hours, followed by treatment with GnRHa (10(-7) mol/L) or GnRHant (10(-6) mol/L) for 0, 5, 15, 30, 60 and 90 minutes, with the 0 min group as the control. Then the protein levels of phosphorylated ERK (p-ERK) and phosphorylated p38 (p-p38) were detected by Western blot, and that of p-ERK determined by the same means after co-incubation of GnRHa or GnRHant with the PKC inhibitor GF109203X at 1, 5, 10 and 20 micromol/L.

RESULTSAfter stimulation of the Leydig cells with GnRHa or GnRHant for different times, the protein level of p-p38 showed no significant difference from that of the control group (P > 0.05). Then the Leydig cells were treated with GF109203X at different concentrations for 20 minutes and with addition of GnRHa for another 10 minutes. The level of p-ERK was significantly decreased (P < 0.05) by GF109203X at 10 and 20 micromol/L. Compared with the control, the p-ERK expression was increased by 65% at 15 minutes (P < 0.05) in the GnRHant stimulation group, by 81% (to the peak) at 30 minutes (P < 0.05), began to fall at 60 minutes, and returned to the base level at 90 minutes. The p-ERK level exhibited no significant difference from that of the control (P > 0.05) after treatment of the Leydig cells with different concentrations of GF109203X for 20 minutes and then with GnRHant for 30 minutes.

CONCLUSIONThe ERK MAPK activation induced by GnRHa depends on the PKC pathway, but not that induced by GnRHant. The p-38 MAPK pathway may not be involved in the effect of GnRH analogues on rat Leydig cells.

Animals ; Cells, Cultured ; Gonadotropin-Releasing Hormone ; analogs & derivatives ; pharmacology ; Leydig Cells ; drug effects ; metabolism ; MAP Kinase Signaling System ; drug effects ; Male ; Rats ; Rats, Sprague-Dawley

OBJECTIVEGonadotropin releasing hormones (GnRH) regulate the expression of annexin 5 in Leydig cells, and annexin 5 is supposed to be a signal molecule in regulating testosterone secretion. This study aimed to investigate the function of annexin 5 in male reproduction by observing its effect on human sperm motility in vitro.

METHODSThe encoding sequence of rat annexin 5 was chemically synthesized and inserted into the HIS fusion expression vector pET28a. The expression of the fusion protein HIS-annexin 5 was induced by isopropyl-beta-D-thiogalactoside (IPTG) under the control of the T7 promoter, and the products were purified by affinity chromatography. The anticoagulant activity of annexin 5 was determined by the modified activated partial thromboplastin time (APTT) test. Semen samples from 15 donors were assigned to a control and an annexin 5 group, the latter treated with recombinant annexin 5 at the concentration of 10(-8) mol/L. Sperm motility and the percentage of grade a + b sperm were measured by computer-assisted semen analysis (CASA) after 20 and 60 min exposure, and the sperm ascending experiment was done after 20 min treatment.

RESULTSThe product of the synthesized target gene was 947 bp in length, and the inserted sequence corresponded to the published encoding sequence of rat annexin 5. The plasmid pET28a-annexin 5 was transformed into E. coli BL21(DE3) and IPTG induced a fusion protein with a relative molecular weight of about 36,000, a purity of 95% and a high anticoagulant activity. Compared with the control group, sperm motility and the percentage of grade a + b sperm were increased by 40% (P < 0.01) and 21% (P < 0.01), respectively, after 20 min treatment with annexin 5, but neither showed any significant improvement after 60 min. The sperm ascending altitude was remarkably elevated after annexin 5 treatment, with extremely significant difference from the control group (37.84 +/- 6.35 vs. 49.5 +/- 12.27, P < 0.01).

CONCLUSIONAn annexin 5 recombinant expression vector was successfully constructed. The protein annexin 5 can be efficiently expressed in E. coli and effectively improve human sperm motility in vitro.

Animals ; Annexin A5 ; genetics ; pharmacology ; Genetic Vectors ; Humans ; Male ; Plasmids ; Rats ; Recombinant Fusion Proteins ; genetics ; pharmacology ; Sperm Motility

BACKGROUNDDiabetes mellitus is associated with coronary dysfunction, contributing to a 2- to 4-fold increase in the risk of coronary heart diseases. The mechanisms by which diabetes induces vasculopathy involve endothelial-dependent and -independent vascular dysfunction in both type 1 and type 2 diabetes mellitus. The purpose of this study is to determine the role of vascular large conductance Ca(2+)-activated K(+) (BK) channel activities in coronary dysfunction in streptozotocin-induced diabetic rats.

METHODSUsing videomicroscopy, immunoblotting, fluorescent assay and patch clamp techniques, we investigated the coronary BK channel activities and BK channel-mediated coronary vasoreactivity in streptozotocin-induced diabetic rats.

RESULTSBK currents (defined as the iberiotoxin-sensitive K(+) component) contribute (65 ± 4)% of the total K(+) currents in freshly isolated coronary smooth muscle cells and > 50% of the contraction of the inner diameter of coronary arteries from normal rats. However, BK current density is remarkably reduced in coronary smooth muscle cells of streptozotocin-induced diabetic rats, leading to an increase in coronary artery tension. BK channel activity in response to free Ca(2+) is impaired in diabetic rats. Moreover, cytoplasmic application of DHS-1 (a specific BK channel b(1) subunit activator) robustly enhanced the open probability of BK channels in coronary smooth muscle cells of normal rats. In diabetic rats, the DHS-1 effect was diminished in the presence of 200 nmol/L Ca(2+) and was significantly attenuated in the presence of high free calcium concentration, i.e., 1 mmol/L Ca(2+). Immunoblotting experiments confirmed that there was a 2-fold decrease in BK-b(1) protein expression in diabetic vessels, without altering the BK channel α-subunit expression. Although the cytosolic Ca(2+) concentration of coronary arterial smooth muscle cells was increased from (103 ± 23) nmol/L (n = 5) of control rats to (193 ± 22) nmol/L (n = 6, P < 0.05) of STZ-induced diabetic rats, reduced BK-b(1) expression made these channels less sensitive to intracellular Ca(2+), which in turn led to enhanced smooth muscle contraction.

CONCLUSIONSOur results indicated that BK channels are the key determinant of coronary arterial tone. Impaired BK channel function in diabetes mellitus is associated with down-regulation of BK-b(1) expression and reduction of the b(1)-mediated BK channel activation in diabetic vessels.

Animals ; Blotting, Western ; Coronary Vessels ; metabolism ; Diabetes Mellitus, Experimental ; metabolism ; physiopathology ; Diabetes Mellitus, Type 1 ; metabolism ; physiopathology ; Electrophysiology ; Large-Conductance Calcium-Activated Potassium Channels ; metabolism ; Male ; Muscle, Smooth, Vascular ; metabolism ; Rats ; Rats, Sprague-Dawley

Objective:To observe the effect of Tongxie Yaofang on the expressions of colon serotonin transporter （SERT）， liver 5-hydroxytryptamine_2A receptor （5-HT_2AR） protein， serum 5-HT and inflammatory factors in ulcerative colitis （UC） model rats of liver stagnation and spleen deficiency， in order to explore the basis of syndrome of liver stagnation and spleen deficiency and the intervention mechanism of Tongxie Yaofang. Method:Fifty male SD rats were randomly divided into blank control group， model group， high， medium and low-dose Tongxie Yaofang group （10，5，2.5 g·kg^-1）， and salazosulacil group （0.3 g·kg^-1）. The ulcerative colitis model of liver depression and spleen deficiency was established by 2，4，6-trinitrobenzene sulfonic acid （TNBS）/ethanol solution enema + restraint stress + diet loss. After successful modeling， the samples were collected after 21 days of drug intervention. Htoxylin eosin （HE） staining and oil red staining were used to observe the pathological changes of colon and liver in each group. Serum interleukin-6 （IL-6）， IL-9， 5-HT and superoxide dismutase （SOD） were detected by enzyme linked immunosorbent assay （ELISA）. Protein expressions of SERT in the colons and 5-HT_2AR in liver of rats were detected by Western blot. Result:Compared with the normal group， obvious ulcers were formed in the colon and lipid droplets in the liver increased in the model group， serum levels of IL-6， IL-9 and 5-HT in the model group increased， while the level of SOD decreased （P<0.05）. The protein expression of SERT in colon decreased， whereas the protein expression of 5-HT_2AR in liver increased （P<0.05）. Compare with model group， the pathological damage of colon was improved， and the formation of lipid droplets in liver was reduced in high， medium-dose Tongxie Yaofang groups and sulfasalazine group. The serum levels of IL-6， IL-9 and 5-HT decreased， while the level of SOD increased in Tongxie Yaofang group and sulfasalazine group （P<0.05）. The protein expression of SERT in colon increased in high，low-dose Tongxie Yaofang groups and sulfasalazine group， and the protein expression of 5-HT_2AR in liver decreased in medium， low dose Tongxie Yaofang groups and sulfasalazine group （P<0.05）. Conclusion:Tongxie Yaofang may reduce the content of 5-HT， and regulate the intestinal motility and sensory system by up-regulating the expression of SERT in the colon， inhibit the expressions of IL-6，IL-9 and other inflammatory factors， and play an anti-inflammatory role， reduce the content of 5-HT and the expression of 5-HT_2AR in the liver， increase the level of SOD， regulate emotion and lipid metabolism in the liver， and then exert the intervention effect on ulcerative colitis with liver depression and spleen deficiency on the whole.