Objective： This study was designed to investigate the relationship between p16 protein expression and gastric carcinogenesis,depth of invasion, lymph node metastasis, and evaluate the role of deletion and mutation of p16 gene in exon 2 in gastric carcinoma. Methods： p16 protein expression in gastric carcinoma and precancerous lesion was examined by streptavidin-peroxidase conjugated(S-P) method; The deletion and mutation of p16 gene were examined respectively by polymerase chain reaction(PCR) and polymerase chain reaction single-strand conformation polymorphism analysis(PCR-SSCP) in gastric carcinoma. Results： ① The positive rates of p16 protein expression were 96.25％ (77/80) in normal gastric mucosa, 92.00％ (45/50) in dysplastic gastric mucosa, and 47.54％ (58/122) in gastric carcinoma. The positive rate of p16 protein expression in gastric carcinoma was significantly lower than that in normal gastric mucosa and in dysplastic gastric mucosa (P<0.05). ② The positive rate of p16 protein expression in mucoid carcinoma (10.00％ ,1/10) was significantly lower than that of poorly differentiated carcinoma (51.22％ ,21/41), undifferentiated carcinoma (57.69％ ,15/26), and signet ring cell carcinoma (62.50％ ,10/16) (P< 0.05). ③ The positive rates of p16 protein in 30 cases paired primary and lymph node metastatic gastric carcinoma were 46.67 ％ (14/30) in primary gastric carcinoma,16.67％ (5/30) in lymph node metastatic gastric carcinoma. The positive rate of lymph node metastatic carcinoma was significantly lower than that of primary carcinoma(P<0.05). ④ Evaluation of mutation and deletion of p16 gene： There was no mutation of p16 gene in exon 2, but there were 5 cases displayed deletion of p16 gene in exon 2 in the 25 primary gastric carcinoma. Conclusions： ① The expression loss of p16 protein is related to carcinogenesis, histopathological subtypes,and lymph metastasis of gastric carcinoma. ② The mutation of p16 gene in exon 2 may not be involved in gastric carcinogenesis. But the deletion of p16 gene in exon 2 might be involved in gastric carcinogenesis.

Animals ; Cells, Cultured ; DNA ; analysis ; Flow Cytometry ; methods ; Fluorescence ; G1 Phase ; Resting Phase, Cell Cycle

OBJECTIVETo evaluate the clinical efficacy of autologous semitendinosus and gracilis tendon grafting with anchor repair for the treatment of chronic achilles tendon rupture and severe scarring.

METHODSFrom April 2010 to October 2012,26 patients with chronic achilles tendon rupture(with Myerson type III ) and severe scarring were treated with autologous semitendinosus and gracilis tendon grafting with anchor repair. There were 19 males and 7 females,with an average age of 32 years old (ranged, 22 to 47 years). The time from injury to surgery was from 3 to 12 months (7 months on average). The plantar flexion strength of all injuried feet attenuated and single heel rise test were positive in 26 cases before operation. Plaster immobilization and routine rehabilitation therapy were performed after operation. Clinical effects were evaluated by Arner-lindholm criterion and complications were observed after operation.

RESULTSAll the patients were followed up from 12 to 24 months with a mean of 16 months. No complications such as achilles tendon re-rupture, wound infection, etc were found during follow-up period. According to the Arner-Lindholm standard, 15 cases got excellent results and 11 good.

CONCLUSIONUsing autologous semitendinosus and gracilis tendon grafts with anchor repair to treat chronic achilles tendon rupture and severe scarring is a perfect surgical procedure.

Achilles Tendon ; injuries ; surgery ; Adult ; Chronic Disease ; Cicatrix ; surgery ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Rupture ; Young Adult

OBJECTIVETo study the morphologic classification of mammary ductal hyperplasia, and its criteria and the significance in distinguishing atypical hyperplasia from carcinoma-in-situ.

METHODSThe clinicopathologic features of 300 cases of hyperplasia of breast were reviewed. Whole-organ H&E sections were also available in 86 cases of breast carcinoma. The occurrence of atypical hyperplasia in adjacent breast tissue was assessed.

RESULTSFibroadenomatoid changes were typically observed in the 21-30 age groups and atypical hyperplasia occurred more frequently in 40-60 age groups. Amongst the hyperplastic cases, cystic diseases of the breast were noted in only 6%. In contrast, fibroadenomatoid changes were more common (25.4%). Atypical ductal hyperplasia occurred in adjacent breast tissue of 65.1% of the carcinoma cases. The incidence was higher (74.9%) if the main lesion was ductal carcinoma-in-situ.

CONCLUSIONSThere is a close association between atypical hyperplasia and breast carcinoma. It is prudent to distinguish between usual and atypical hyperplasia. Morphologic differentiation between atypical ductal hyperplasia and ductal carcinoma-in-situ may sometimes be difficult.

Adult ; Breast ; pathology ; Breast Neoplasms ; pathology ; Carcinoma in Situ ; pathology ; Carcinoma, Ductal, Breast ; pathology ; Carcinoma, Intraductal, Noninfiltrating ; pathology ; Diagnosis, Differential ; Female ; Fibroadenoma ; pathology ; Humans ; Hyperplasia ; pathology ; Middle Aged ; Precancerous Conditions ; pathology

OBJECTIVETo investigate the role of Runx3 gene CpG island methylation in the development of human gastric carcinoma.

METHODSA total of 150 tumor specimens from patients with gastric carcinoma and 50 normal tissue specimens were selected. Methylation specific PCR (MSP) and pyrosequencing (PS) were used to detect the methylation status of Runx3 gene promoter.

RESULTSCompared to normal tissue samples, a significant increase of CpG island methylation status of Runx3 gene was observed in gastric carcinomas (MSP: 67.3% vs. 40.0%, P = 0.002; PS: 76.0% vs. 30.0%, P < 0.01). Runx3 gene methylation was only related to tumor size (P < 0.05) based on MSP analysis. PS test however showed that the extent of methylation of Runx3 gene was related to the tumor size (P = 0.004), Lauren's classification (P = 0.043), depth of invasion (P < 0.01), lymph node metastasis (P = 0.021) and TNM staging (P = 0.045).

CONCLUSIONSMethylation status of Runx3 gene detectable by PS is closely correlated with clinicopathological parameters of gastric carcinoma, including tumor size, Lauren's classification, depth of invasion, lymph node metastasis and TNM staging. PS is more sensitive than MSP in the detection of Runx3 gene methylation, which may serve as an important marker for early diagnosis and treatment of gastric carcinoma.

Adult ; Aged ; Aged, 80 and over ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; CpG Islands ; genetics ; DNA Methylation ; Disease-Free Survival ; Female ; Follow-Up Studies ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Polymerase Chain Reaction ; Promoter Regions, Genetic ; genetics ; Sequence Analysis, DNA ; Stomach Neoplasms ; genetics ; metabolism ; pathology ; Tumor Burden ; Young Adult

BACKGROUNDRecent research suggests that beta(2)-adrenergic agonists increase alveolar fluid clearance (AFC) under physiologic and pathologic conditions. It is unknown whether beta(3)-adrenergic agonists also increase AFC under pathologic conditions. The aim of this study was to investigate the effect of beta(3)-adrenergic agonists on AFC following hypoxic lung injury and the mechanisms involved.

METHODSHypoxic rats were exposed to 10% oxygen. BRL-37344 (beta(3)-adrenergic agonist) or CGP-12177 (selective beta(3)-adrenergic agonist) alone or combined with beta receptor antagonists, sodium channel blockers, or Na(+)/K(+)-ATPase blockers were perfused into the alveolar space of rats exposed to 10% oxygen for 48 hours. Total lung water content (TLW) and AFC were measured.

RESULTSAFC did not change for the first 24 hours but then decreased after 48-hour exposure to 10% oxygen. The perfusion of BRL-37344 or CGP-12177 significantly increased AFC in normal and hypoxic rats. The AFC-stimulating effect of CGP-12177 was lowered with amiloride (a Na(+) channel blocker) and ouabain (a Na(+)/K(+)-ATPase inhibitor) by 37% and 49%, respectively. Colchicine significantly inhibited the effect of CGP-12177.

CONCLUSIONSThese findings suggest that beta(3)-adrenergic agonists can increase AFC during hypoxic lung injury in rats and accelerate the amelioration of pulmonary edema.

Adrenergic beta-Agonists ; therapeutic use ; Animals ; Body Fluids ; drug effects ; metabolism ; Ethanolamines ; therapeutic use ; Hypoxia ; physiopathology ; Male ; Propanolamines ; therapeutic use ; Pulmonary Alveoli ; drug effects ; metabolism ; pathology ; Pulmonary Edema ; drug therapy ; etiology ; metabolism ; Rats ; Rats, Wistar

OBJECTIVETo investigate the effect of phenylephrine (an α-adrenergic agonist) on alveolar fluid clearance (AFC) in ventilator-induced lung injury and the possible mechanism involved.

METHODSA total of 170 male Wistar rats were randomly allocated into 17 groups (n=10) using random number tables. Short-term (40 minutes) mechanical ventilation with high tidal volume (HVT) was performed to induce lung injury, impair active Na+ transport and lung liquid clearance in the rats. Unventilated rats served as controls. To demonstrate the effect of phenylephrine on AFC, phenylephrine at different concentrations (1×10(-5), 1×10(-6), 1×10(-7), 1×10(-8), and 1×10(-9) mol/L) was injected into the alveolar space of the HVT ventilated rats. To identify the influence of adrenergic antagonists, Na(+) channel, and microtubular system on the effect of phenylephrine, phenylephrine at 1×10(-5) mol/L combined with prazosin (an α1-adrenergic antagonist, 1×10(-4) mol/L), yohimbine (an α2-adrenergic antagonist, 1×10(-4) mol/L), atenolol (a β1- adrenergic antagonist, 1×10(-5) mol/L), ICI-118551 (an β2-adrenergic antagonist, 1×10(-5) mol/L), amiloride (a Na+ channel blocker, 5×10(-4) mol/L), ouabain (a Na(+)/K(+)-ATPase blocker, 5×10(-4) mol/L), colchicine (a microtubular disrupting agent, 0.25 mg/100 g body weight), or β-lumicolchicine (an isomer of colchicine, 0.25 mg/100 g body weight) were perfused into the alveolar space of the rats ventilated with HVT for 40 minutes. AFC and total lung water content were measured.

RESULTSBasal AFC in control rats was (17.47±2.56)%/hour, which decreased to (9.64± 1.32)%/hour in HVT ventilated rats (P=0.003). The perfusion of phenylephrine at 1×10(-8), 1×10(-7), 1×10(-6), and 1×10(-5) mol/L significantly increased the AFC in HVT ventilated rats (all P<0.05). This effect of phenylephrine on AFC was suppressed by prazosin, atenolol, and ICI-118551 in HVT ventilated rats by 53%, 31%, and 37%, respectively (all P<0.05). The AFC-stimulating effect of phenylephrine was lowered by 33% and 42% with amiloride and ouabain, respectively (both P<0.05). Colchicine significantly inhibited the effect of phenylephrine (P=0.031).

CONCLUSIONPhenylephrine could increase the AFC in HVT-ventilated rats and accelerate the absorption of pulmonary edema.

Animals ; Male ; Phenylephrine ; therapeutic use ; Pulmonary Alveoli ; metabolism ; Rats ; Rats, Wistar ; Sodium-Potassium-Exchanging ATPase ; metabolism ; Ventilator-Induced Lung Injury ; drug therapy ; metabolism ; pathology

OBJECTIVETo evaluate the effects of portaazygous disconnection (PAD), portacaval shunt (PCS) and distal splenocaval shunt (DSCS) on the portosytemic shunting (PSS), hepatic function (HF), hepatic mitochondrial respiratory function (HMRF), oral glucose tolerance test (OGTT) and arterial ketone body ratio (KBR) in order to provide a sound basis for selecting suitable operations for patients.

METHODSUsing a cirrhotic portal hypertensive model induced by CCl4/ethanol in Wistar rats, the PSS, HF, HMRF, OGTT and KBR were determined three weeks after PCS, DSCS and PAD.

RESULTSIt was revealed that: (1) In the cirrhotic portal hypertension rats, the PSS increased significantly, HMRF and hepatic reserve function (HRF) decreased significantly when compared with the control rats. (2) At the time of first postoperative week, the mean blood glucose value in the 120-minute OGTT in each PAD, PCS and DSCS groups had significant differences compared with the cirrhotic control group. But during the second and third postoperative weeks, the mean blood glucose values in the 120-minute OGTT in both PAD and DSCS groups had no significant differences compared with the cirrhotic control group except for the PCS group. The values of KBR in the three operative groups decreased significantly compared with the cirrhotic control group during the two postoperative weeks. In the third postoperative week, only the values of KBR in the PCS group had a significant difference compared with the cirrhotic control group. (3) After PCS, the PSS was further increased; HF and HMRF were significantly decreased. Little improvement was found in the third postoperative week. (4) After DSCS and PAD, the above mentioned indices were less influenced, and they were restored more quickly than those in the PCS group.

CONCLUSIONWe found that PAD and DSCS are more desirable than PCS.

Animals ; Hypertension, Portal ; etiology ; surgery ; Liver Cirrhosis, Experimental ; complications ; surgery ; Portacaval Shunt, Surgical ; Portasystemic Shunt, Surgical ; methods ; Rats ; Rats, Wistar

OBJECTIVETo ascertain the pathogen of aseptic encephalitis epidemic in Long-Yan city in Fujian, and to find out the genetic characteristics of the virus.

METHODSRapid detection of enteroviral RNA by reverse transcription polymerasechain reaction (RT-PCR) was directly carried out in cerebrospinal fluid(CSF) to isolate and identify the viruses from CSF at the same time, and to detect the neutralization antibody in two serum specimens collected in acute and convalescence phase. Nucleotides of VP1 region was also analyzed by constructing phylogenetic tree.

RESULTSECHO 19 infection was rapidly diagnosed and sequence analysed by RT-PCR, and then echovirus type 19 from 16 of 30 CSF samples (53.33%) was isolated and detected using RD and Hep-2 cells simultaneity. The titer of ECHO 19 neutralization antibody became positive or increased by 4 times from acute to convalescence phase in 4 of the 5 patients. Phylogenetic analyses of the VP1 genes of these isolates showed that their nucleotides identity were 98.9% -100.0% which were different from those ECHO 19 from GeneBank database by 13.0%-22.4%.

CONCLUSIONThe etiology of the epidemic of aseptic encephalitis was attributed to ECHO 19. The method of molecular identification not only provided rapid diagnosis of enterovirus infections, but also information about the genetic character of the viruses.

Antibodies, Neutralizing ; analysis ; China ; epidemiology ; Echovirus Infections ; diagnosis ; immunology ; virology ; Encephalitis, Viral ; epidemiology ; virology ; Enterovirus B, Human ; genetics ; isolation & purification ; Humans ; Phylogeny ; Reverse Transcriptase Polymerase Chain Reaction

The study was aimed to detect the expressions of costimulatory molecules on CD3(+)CD4(+) T cells so as to accumulate informations for investigation of mechanism of myelodysplastic syndrome. 11 healthy blood donors as control and 38 patients with MDS de novo were studied. 38 MDS patients were divided into RA/RARS group and RAEB/RAEB-t group according to FAB classification. The expressions of CD28, CD154, CTLA-4, PD-1, CD25 on CD3(+)CD4(+) T cells in peripheral blood were detected by FCM. The results indicated that as compared with normal controls, the expression of CD28 in MDS patients decreased, and CD154 increased. The percentages of CTLA-4, PD-1 and CD25 in MDS were obviously higher than that in normal controls; the differences of CTLA-4, PD-1 and the ratio of CTLA-4/CD28 between RAEB/RAEB-t and RA/RARS were more significant with progressing of MDS. In conclusion, the expressions of costimulatory molecule in MDS patients were abnormal, which may be involved in the pathogenesis of MDS.
Adult ; Antigens, CD ; metabolism ; Apoptosis Regulatory Proteins ; metabolism ; CD28 Antigens ; metabolism ; CD40 Ligand ; metabolism ; CTLA-4 Antigen ; Case-Control Studies ; Female ; Humans ; Interleukin-2 Receptor alpha Subunit ; metabolism ; Lymphocyte Count ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; metabolism ; Programmed Cell Death 1 Receptor ; T-Lymphocytes ; immunology ; metabolism ; Young Adult