Objective:To investigate the association between p53 codon 72 polymorphism and the prognosis of breast cancer pa-tients receiving chemotherapy and radiotherapy after surgery. Methods:A total of 427 breast cancer patients treated with chemo-radio-therapy after surgery at Beijing Cancer Hospital were selected for this study. Polymerase chain reaction–restriction fragment length polymorphism was adopted to analyze the p53 codon 72 polymorphism. Survival analysis was conducted to compare the disparities of recurrence and survival among the patients with different p53 codon 72 polymorphic variants. Results:The distribution of three geno-types of p53 codon 72 in our cohort is as follows:Pro/Pro 18.3%(78/427), Pro/Arg 44.0%(188/427), and Arg/Arg 37.7%(161/427). No significant difference was observed among the local recurrence-free survival (LRFS), loco-regional recurrence-free survival (LR-RFS), distant disease-free survival (DDFS), and overall survival (OS) among the three genotypes (all P>0.05). Among the 303 estro-gen receptor (ER)-positive patients, OS was significantly better in patients with Arg/Arg genotype than those with Pro/Pro genotype (χ2=6.33, P=0.042). The multivariate analysis showed that the p53 codon 72 polymorphism is an independent factor of prognosis for LRFS, LRRFS, DDFS, and OS of ER-positive patients. For the ER positive patients with Pro/Pro genotype, the local recurrence, local-regional recurrence, distant metastasis, and mortality risks were 5.9 (HR=5.9, 95%CI 1.1-31.1, P=0.036), 3.1 (HR = 3.1, 95% CI 1.1-9.1, P=0.039), 2.8 (HR=2.8, 95% CI 1.3-6.0, P=0.010), and 4.0 (HR=4.0, 95% CI 1.3-12.0, P=0.013) times higher than those with Arg/Arg genotype, respectively. Conclusion:For ER-positive breast cancer patients who underwent surgery and chemo-radiotherapy, the local recurrence, loco-regional recurrence, distant metastasis, and mortality risk with Pro/Pro genotype are significantly higher compared to those with Arg/Arg genotype.

Objective To establish a lineage tracing method with Lgr5-EGFP-CreERT2/ROSA26-stop-EYFP mouse for observing the role of Lgr5⁺ intestinal stem cells in the renovation of small intestine tissue, and investigate the effects of radiation on the reconstruction of small intestinal stem cell tissue. Methods-Lgr5-EGFP-CreERT2/ROSA26-stop-EYFP mice were identified by genotype analysis. Tamoxifen was used to induce the expression of fluorescent protein in mice. The differentiation of intestinal stem cells induced by Tamoxifen was observed by laser confocal microscopy. Lgr5-EGFP-CreERT2/ROSA26-stop-EYFP mice were irradiated with⁶⁰Co γ-rays, and the effects of irradiation on the reconstruction of small intestinal stem cell tissue were examined. Results-Double positive mice were obtained by identification of Lgr5 gene (174bp) in DNA extraction from the mice tail with PCR method. In mice treated with single injection of Tamoxifen (100mg/kg), it was observed by laser confocal scanning microscopy that the Lgr5⁺ intestinal stem cells started dividing at the 5th day after inducement, reached the top of the villi at the 7th day, fluorescence appeared in a few of whole intestinal villi at the 14th day, and spread over more intestinal villi with dense fluorescence at the 45th day. However, in mice exposed to 15Gy irradiation, the intestinal villi fell off seriously without fluorescence in crypts. Conclusion-The lineage tracing model of intestinal stem cell has been successfully established and then used to evaluate the irradiation injuries to intestinal stem cells.

Objective The immune hypofunction of the red blood cell on maintenance hemodialysis patients is one main reason of tumor,and oxidative stress is proved to be involved in decrease of existence quality and the occurrence of various complications of patients with hemodialysis.To investigate the status of erythrocyte immune and high oxidative stress of the hemodialysis patients,as well as the effect of Huangqifuzitang on immune function of red blood cell and oxidative stress in patients with hemodialysis.Methods Twenty healthy people were chosen as control group,and 20 patients were selected as the treatment group.CD58,CD59,superoxide dismutase(SOD),malonaldehyde(MDA) were measured before hemodialysis and after 12 days administration of Huangqifuzitang in the treatment group,and compared with those of the control group.Results There was significant difference between before and after taking Huangqifuzitang in treatment group and control group in terms of CD58 ((38.02 ±8.98) vs.(47.39 ±7.78) vs.(59.10±4.59),F=4.506,P=0.000),CD59 ((62.69 ± 20.84) vs.(80.95 ± 20.42) vs.(193.86 ± 19.87) ; F =239.347,P =0.000),SOD ((68.09 ± 11.86) vs.(78.73±10.58) vs.(111.09±16.61) kU/L;F=21.318,P=0.000),MDA((5.98±2.06) vs.(4.54 ±0.62) vs.(3.03 ± 1.10) μmoL/L;F =55.359,P =0.000) levels.Before taking Huangqifuzitang,the concentration of CD58,CD59 and SOD in treatment group was significant decreased compared with control group,but MDA significant increased,and there was significant difference (P ＜ 0.01).After 12 days administration of Huangqifuzitang in the treatment group,the concentration of CD58,CD59 and SOD was significant increased compared with before taking Huangqifuzitang,but MDA significant significant decreased,and there was significant difference (P ＜ 0.01 or P ＜ 0.05) Conclusion The red blood cell surface receptors CD58,CD59 decreased and oxidative stress index SOD decreased significantly,MDA increased significantly in hemodialysis patients.Huangqifuzitang was proved to be with the ability of enhancing the immune of red bloodCD58,CD59 and decreasing the oxidative stress level in patients with hemodialysis.

Objective To evaluate the safety of surgical procedures for patients after heart valve prosthesis implantation.Methods Clinical data of 12 cases with heart valve prosthesis implantation undergone other surgical treatment from November 1996 to December 2005 were retrospectively analyzed.All the cases had routine oral warfarin with prothrombin time (PT) of 20.0—28.3 s averaged 23.5 s, international normalized ratio (INR) for prothrombin of 1.79—2.23 averaged 1.95 and heart functional class Ⅰ—Ⅲ.Among them,appendectomy was performed in three cases with acute appendicitis,reposition and repair in one with inguinal hernia,radical gastrectomy in two with gastric carcinoma,left hemicolectomy in one,cholecystectomy in three,left femoral head replacement in one,and bilateral high ligation and ablation of great saphenous vein in one.Elective surgical operation was performed in seven cases,and emergency operation in five.In those with elective surgery,warfarin was stopped 2—3 days before operation,while 5—10 mg vitamin K_1 was injected intramuscularly 6—8 hours before emergency surgery with preoperative median PT of 15.1 and 15.3 s and median INR of 1.24 and 1.30,respectively.In operation,5—10 mg vitamin K_1 were injected intravenously into the patients by drip depending on their bleeding on the surface of wound.ECG,blood pressure,hemoglobin and oxygen saturation were routinely monitored for all the cases intraoperatively and postoperatively.For the cases with heart function above class Ⅱ,fluid infusion was adjusted based on intubated central venous pressure,and for those with general anesthesia,analyses of blood gases and electrolyte were monitored routinely in operation.Results OPeration time averaged 20—160 rain in all the 12 patients,with blood loss 5—280 ml in average and without complications of massive hemorrhage,thrombosis and heart failure.Conclusions Surgical operation was safe for patients with heart valve prosthesis implantation,if preoperative PT and INR were adjusted to about 15 s and 1.30,respectively by cessation of warfarin or application of vitamin K_1,combined with careful manipulation and strengthened perioperative management.

OBJECTIVETo compare the clinical effectiveness of ropivacaine mesylate and ropivacaine hydrochloride in epidural anesthesia and postoperative analgesia.

METHODSForty-four patients undergoing epidural anesthesia for intraperitoneal hysterectomy or hysteromyomectomy were randomly assigned to two equal groups. Group ropivacaine hydrochloride received 15 ml ropivacaine hydrochloride of 7.5 mg/ml in anesthetic and 2 mg/ml in postoperative analgesic subsequently while group ropivacaine mesylate correspondingly received ropivacaine mesylate of both 15 ml of 8.94 mg/ml and 2.374 mg/ml.

RESULTSNo significant differences were found in the onset, extent, and duration of sensory and motor blockage, and also in the hemodynamic stability, when ropivacaine mesylate was comparable with ropivacaine hydrochloride. No severe adverse events were observed in this study.

CONCLUSIONRopivacaine mesylate is an effective and safe alternative to ropivacaine hydrochloride in epidural anesthesia and postoperative analgesia.

Adolescent ; Adult ; Aged ; Amides ; chemistry ; therapeutic use ; Anesthesia, Epidural ; Female ; Humans ; Hysterectomy ; Middle Aged ; Pain, Postoperative ; drug therapy

OBJECTIVETo initially observe the effect of classical endotracheal intubation on endotracheal bacterial contamination and evaluate the validity of protective endotracheal intubation on reducing endotracheal bacterial contamination.

METHODSNinety elective patients undergoing general anesthesia for hysterectomy were randomly assigned to two equal groups. Group II received endotracheal intubation protected by sterilized transparent sleeve while group I correspondingly adopted unprotective classical endotracheal intubation. Endotracheal swab sampling and bacterial counting were performed on the principle of aseptic processing before endotracheal intubation and extubation, respectively.

RESULTSBacteria were found in 62 of 180 samples. The difference of bacterial counting between before extubation and before intubation was (-0.3 +/- 35.6) 100 CFU/ ml in group II, lower than that in group I, which was (21.4 +/- 56.7) 100 CFU/ml (P<0.05).

CONCLUSIONEndotracheal bacterial contamination may be caused by unprotective classical endotracheal intubation and could be reduced by protective endotracheal intubation.

Anesthesia, General ; Bacteria ; isolation & purification ; Female ; Humans ; Hysterectomy ; Intubation, Intratracheal ; adverse effects ; methods ; Trachea ; microbiology

OBJECTIVETo observe the protection of Qingyuan Shenghua Decoction (QSD) on multiple organs of sepsis patients after bone trauma, and to preliminarily explore its mechanism.

METHODSTotally 60 sepsis patients after bone trauma were randomly assigned to the treatment group and the control group according to random digit table, 30 in each group. All patients received routine Western medical treatment. Patients in the treatment group additionally took QSD or were nasally fed with QSD, one dose per day for 1 week. Changes of WBC, oxygenation index (PaO2/FiO2), serum creatinine (SCr), total bilirubin (TBIL), aspartate aminotransferase (AST), fibrinogen (FIB), D-dimer (DD), activated partial thromboplastin time (APTT), pro-calcitonin (PCT), C-reactive protein (CRP), heart rate (HR), mean arterial pressure (MAP), intra-abdominal pressure, scores for Acute Physiology and Chronic Health Evaluation II (APACHE II), sequential organ failure assessment (SOFA) scores were observed before treatment and on day 1, 3 and 7 after treatment.

RESULTSCompared with the control group at the same time point, MAP increased at post-treatment day 1 and 3; CRP, APTT, HR, SCr, TBIL, AST, intra-abdominal pressure at post-treatment day 3 obviously decreased in the treatment group (P < 0.05, P < 0.01). WBC, SOFA scores, PCT, CRP, APACHE II, APTT, D-D, HR, SCr, TBIL, AST and intra-abdominal pressure significantly decreased; FIB, MAP and PaO2/FiO2 obviously increased at post-treatment day 7 (P < 0.05, P < 0.01).

CONCLUSIONQSD had good protective effect on multiple organ function in sepsis patients after bone trauma, and its mechanism might be related with effectively clearing endotoxin, alleviating inflammatory reactions, and fighting against coagulation dysfunction.

APACHE ; Blood Coagulation ; Bone Diseases ; complications ; C-Reactive Protein ; metabolism ; Calcitonin ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; therapeutic use ; Fibrin Fibrinogen Degradation Products ; metabolism ; Humans ; Inflammation ; Partial Thromboplastin Time ; Protein Precursors ; metabolism ; Sepsis ; drug therapy ; etiology

OBJECTIVEThe aim of this study was to evaluate the effects of XRCCl gene polymorphisms and its haplotype on the susceptibility of pancreatic carcinoma.

METHODSPeripheral blood DNA was extracted from 210 pancreatic carcinoma patients and 213 control subjects. SNaPshot technique was used for genotyping seven SNP sites of the XRCCl gene (rs3213403, rs25487, rs1799782, rs731420, rs1001581, rs12611088, and rs3213282). Logistic regression model was performed to analyze the relationship of different genotypes or haplotype and the susceptibility of pancreatic carcinoma.

RESULTSThe frequency for allele A at site rs25487 in the case group was significantly higher than that in the control group (P < 0.05). The frequency of GG, GA and AA genotype between the case group and control group had statistically significant differences (P < 0.05). Compared with GG genotype, the risk of pancreatic carcinoma in the subjects carrying mutated allele A (GA+AA) was increased by 0.648 times (P < 0.05). Among them the pancreatic carcinoma risk of individuals carrying A allele was increased by 0.552 times compared with the individuals carrying G allele. The frequency of allele and genotype at site rs1799782 in the case group and control group had a significant difference (P < 0.05). Compared with the CC genotype, the risk of pancreatic carcinoma in the subjects carrying mutated allele T (CT+TT) was increased by 0.683 times. Among them the pancreatic carcinoma risk of individuals carrying T allele was increased by 0.549 times compared with the individuals carrying C allele. Significant differences were observed in linkage disequilibrium between any two of the seven SNPs (P < 0.05), the frequency of H4-AGCCCGC, H6-GGCCCGG or H7-AGCCTAG haplotypes was significantly lower in the case group than that in the control group (P < 0.05).

CONCLUSIONSThe single nucleotide polymorphisms of rs25487 and rs1799782 for XRCC1 gene may be correlated with the occurrence of pancreatic carcinoma. The haplotypes of H4-AGCCCGC, H6-GGCCCGG and H7-AGCCTAG might be a potential genetic protective factor for the occurrence of pancreatic carcinoma.

Alleles ; DNA-Binding Proteins ; genetics ; metabolism ; Genetic Predisposition to Disease ; epidemiology ; Genotype ; Haplotypes ; Humans ; Pancreatic Neoplasms ; epidemiology ; Polymorphism, Single Nucleotide ; X-Rays ; X-ray Repair Cross Complementing Protein 1

This study, using cultured bovine pulmonary artery endothelial cells (BPAECs), was undertaken to investigate the roles of endogenous ONOO(-) in LPS-caused injury in endothelial cells. The fluorescent intensity of nitrotyrosine (NT), a specific marker of ONOO(-) generation, in BPAECs represented the content of endogenous ONOO(-) generation. The fluorescent intensity of NT and the number of NT positive cells were detected with flow cytometry (FCM), and the percentage of NT positive cells was calculated. The results are as follows. (1) LPS (1, 5 and 10 microg/ml) caused a marked increase in fluorescent intensity of NT in a dose-dependent manner, which was significantly increased compared to the vehicle group (P<0.01).The number and percentage of NT positive cells were markedly increased (both P<0.05 vs vehicle group). Aminoguanidine (AG), a selective inhibitor of inducible nitric oxide synthase (iNOS), inhibited LPS-induced increase in fluorescent intensity of NT in BPAECs. However, the number and percentage of NT positive cells had a tendency to reduce. (2) LPS brought about an enhancement in MDA content and the activity of LDH in cultured supernatant. AG reversed the enhancement in MDA content induced by LPS (P<0.01). In contrast, AG had a marginal effect on the activity of LDH. (3) LPS induced an increase in apoptotic rate in BPAECs in a dose-dependent manner. The number of apoptotic cells markedly increased as well. Some BPAECs stained with fluorescent probe ethidium bromide showed morphological features of apoptosis with chromatin condensation and nuclear fragmentation. AG reduced the apoptotic rate and the number of apoptotic cells, both of which were still higher than those of vehicle group (P<0.05). LPS led to inhibition of mitochondrial respiration and membrane potential in an accumulation manner. In conclusion, LPS caused injury to cultured BPAECs and increased the production of ONOO(-).The cytotoxicity of LPS may be mediated by the endogenous ONOO(-).
Animals ; Cattle ; Cells, Cultured ; Endothelial Cells ; cytology ; metabolism ; pathology ; Lipopolysaccharides ; toxicity ; Lung Injury ; physiopathology ; Peroxynitrous Acid ; biosynthesis ; physiology ; Pulmonary Artery ; cytology ; pathology

OBJECTIVETo observe the effect of sensory neuropeptide substance P combined with epidermal stem cells (ESC) on wound healing and nerve regeneration in diabetic rats.

METHODSESC that had been isolated from SD rats were identified and cultured in vitro, and they were inoculated onto nourishing layer of amniotic membrane to construct amniotic membrane-ESC. Four full-thickness skin wounds were produced on the back of each of 48 diabetic rats. The resulted 192 wounds were randomly divided into ESC + substance P group, ESC group, substance P group, and control group according to the lottery method, with 48 wounds in each group. Wounds in ESC + substance P group and ESC group were transplanted with amniotic membrane-ESC, and those in substance P group and control group were transplanted with amniotic membrane. After transplantation, 250 µL substance P in the concentration of 1 × 10(-7) mol/L was injected around and into the middle of the wounds in ESC + substance P group and substance P group, 2 times a day, and continued for 4 days, while 250 µL PBS solution was injected in the above-mentioned position in ESC group and control group as control, 2 times a day, and continued for 4 days. On post injury day (PID) 4, 7, 10, 14, 17, and 23, the wound healing rate (with 8 wounds at each time point) was observed and determined, and changes in wound tissue structure were observed with HE staining. On PID 4, 7, and 10, collagen distribution in wound tissue was observed with Masson staining, and type I and type III collagen deposition in wound tissue was respectively observed after immunohistochemical staining. The distribution of protein gene product 9.5 (PGP 9.5) and regeneration of substance P positive nerve fibers in wound tissue were observed with immunohistochemical staining on PID 14 and 23. Data were processed with one-way analysis of variance and t test.

RESULTS(1) The wound healing rate in ESC + substance P group reached 100.0% on PID 14, which was obviously earlier than that in ESC group, substance P group, and control group, healing was respectively observed on PID 17, 17, and 23. The wound healing quality in ESC + substance P group was better than that in the other three groups as shown by HE staining. (2) On PID 10, collagen that was darkly stained and widely distributed was observed in wound tissue of ESC + substance P group and substance P group, while collagen in the other two groups was lightly stained and narrowly distributed. Deposition quantity of type I collagen gradually increased, and that of type III collagen gradually decreased in the wounds of each group over time. On PID 4, 7, and 10, distribution amount of type I collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.72, 118.21, 26.71, P values all below 0.01) and control group (with t value respectively 44.37, 22.76, 30.32, P values all below 0.01), while there was no significance between ESC + substance P group and substance P group. On PID 4, 7, and 10, distribution amount of type III collagen in wound tissue of ESC + substance P group was significantly higher than that in ESC group (with t value respectively 32.27, 28.68, 14.51, P values all below 0.01) and control group (with t value respectively 35.68, 22.52, 22.24, P values all below 0.01). (3) A large amount of PGP 9.5 and regeneration of substance P positive nerve fibers, and some peripheral nerve fibers in deep wound extending to epidermis were observed in wound tissue of ESC + substance P group and substance P group. A small amount of PGP 9.5 and regeneration of substance P positive nerve fibers without peripheral nerve fibers extending to epidermis were observed in deep wound tissue of ESC group and control group. On PID 14, 23, ratios of area of PGP 9.5 positive nerve fiber in the wounds of ESC + substance P group were (3.86 ± 0.25)% and (7.03 ± 0.28)%, and they were significantly higher than those of ESC group [(1.48 ± 0.30)%, (3.01 ± 0.43)%, with t value respectively 23.95, 30.27, P values all below 0.01] and control group [(1.46 ± 0.23)%, (2.84 ± 0.29)%, with t value respectively 27.35, 40.32, P values all below 0.01]. On PID 14, 23, ratios of substance P positive nerve fiber area in the wounds of ESC + substance P group were (2.01 ± 0.14)% and (1.19 ± 0.11)%, which were obviously higher than those of ESC group [(0.85 ± 0.17)%, (1.34 ± 0.21)%, with t value respectively 20.50, 2.60, P < 0.05 or P < 0.01] and control group [(0.74 ± 0.15)%, (1.30 ± 0.17)%, with t value respectively 23.98, 2.41, P < 0.05 or P < 0.01].

CONCLUSIONSJoint application of substance P and ESC can effectively promote healing of wound and nerve regeneration in diabetic rats.

Animals ; Diabetes Mellitus, Experimental ; pathology ; Epithelial Cells ; cytology ; Nerve Regeneration ; Rats ; Rats, Sprague-Dawley ; Stem Cells ; cytology ; Substance P ; pharmacology ; therapeutic use ; Wound Healing