Journal of Nanjing Medical University最新文献

Objective

To investigate the effect of hepatic ischemia-reperfusion(I/R) on bile canalicular F-actin microfilaments in rats.

Methods

A rat model of hepatic ischemia-reperfusion was employed and the ischemia time was 35 min. The activity of serum alanine aminotransferase (ALT), aspartate aminotransferase(AST), γ-glutamyl transferase(GGT) and the level of total bilirubin(TBIL) were measured. Changes in the bile canaliculi were observed by transmission electron microscope. The modification of F-actin microfilaments was quantified by using FITC-Phalloidin and analyzed by confocal laser scanning microscopy imaging.

Results

Modifications of F-actin staining were consistent with the observations made by transmission electron microscopy. The staining of F-actin was normal in hepatocytes before reperfusion but decreased significantly after reperfusion, and there was a marked loss of canalicular microvilli after reperfusion, which coincided with abnormal serum GGT and TBIL levels.

Conclusion

Reperfusion, not short-term ischemia, induced a disruption of F-actin microfilaments and a loss of microvilli. These modifications could lead to the impaired bile secretion by damaging canalicular contraction, and could be the main mechanisms of cholestasis after hepatic ischemia-reperfusion in rats.

Objective

To record Calcium, Potassium and Sodium currents in acutely isolated hippocampal pyramidal neurons.

Methods

Hippocampal CA3 neurons were freshly isolated by 1 mg protease/3 ml SES and mechanical trituration with polished pipettes of progressively smaller tip diameters. Patch clamp technique in whole-cell mode was employed to record voltage-gated channel currents.

Results

The procedure dissociated hippocampal neurons, preserving apical dendrites and several basal dendrites, without impairing the electrical characteristics of the neurons. Whole-cell patch clamp configuration was successfully used to record voltage-gated Ca²⁺ currents, delayed rectifier K⁺ current and voltage-gated Na⁺ currents.

Conclusion

Protease combined with mechanical trituration may be used for the dissociation of neurons from rat hippocampus. Voltage-gated channels currents could be recorded using a patch clamp technique.

Objective

To investigate the silencing effects of recombinant adenovirus Ad-shRNA-MK on midkine(MK) gene in pancreatic cancer cells.

Methods

Ad-shRNA-MK was used to infect pancreatic cancer BxPC-3 cells. Assays were conducted for knockdown of the MK gene on the day of infection and on the 1^st, 3^rd, 5^th, 7^th, and 9^th days post-infection by using immunocytochemistry, real-time RT-PCR, and Western blot analysis.

Results

The adenoviral Ad-shRNA-PTN was constructed successfully, and infection was confirmed by electron microscopic observation. By using real-time RT-PCR, the inhibition rates of MK mRNA expression in the BxPC-3 cells were 20%, 80%, 55%, and 23% on the 1^st, 3^rd, 5^th, and 7^th days post-infection. Immunocytochemistry and Western blot analysis confirmed this effect at the gene product level.

Conclusion

Efficient and specific knockdown of MK in pancreatic cancer cells by adenoviral Ad-shRNA-PTN is a potentially powerful tool for the study of gene therapy of pancreatic cancer nerve infiltration.

The review presents the epidemiology of leishmaniasis in the Eastern Africa region. We searched PUB MED and MEDLINE with several key words-namely, “leishmaniasis”;“cutaneous”, “diffuse cutaneous”, “mucosal”, and “visceral leishmaniasis”; “kala azar”, and “post kala azar dermal leishmaniasis”, -for recent clinical and basic science articles related to leishmaniasis in countries in the Eastern Africa region. Poverty, wars, conflicts and migration have significantly aggravated leishmaniases in Eastern Africa. Of particular concern is the increasing incidence of Leishmania-HIV co-infection in Ethiopia where 20∼40% of the persons affected by visceral leishmaniasis are HIV-co-infected. Sudan has the highest prevalence rate of post kala-azar dermal leishmaniasis(PKDL) in the world, a skin complication of visceral leishmaniasis(VL) that mainly afflicts children below age ten. In view of its spread to previously non-endemic areas and an increase in imported cases, leishmaniasis in Eastern Africa should be considered a health emergency.

Objective

To investigate the relation between hepatitis B virus(HBV) genotypes and the double mutation of A-to-T nucleotide(nt) 1762 and G-to-A nt 1764 in basic core promoter(BCP T1762/A1764) in patients of the Li nationality.

Methods

Subjects were 125 HBV DNA positive patients that belong to the Li nationality on Hainan Island. HBV DNA genotype was determined by real time fluorimetry polymerase chain reaction. BCP T1762/A1764 mutation was performed using the direct sequencing method.

Results

The prevalence rates of genotype B, genotype C, genotype D, genotype C and D mixed infection(genotype C + D) and genotype B and D mixed infection (genotype B + C) were 31.20%, 53.60%, 12.00%, 2.40% and 0.80% respectively. Mutation frequencies in patients infected with HBV genotype C(58.21%) were significantly higher than in those infected with other genotypes (P < 0.01). The serum viral load of the patients with genotype C(5.74±1.21) was also higher than that of those with genotype B(P < 0.01).

Conclusion

The major genotypes in the Li nationality were genotype C and genotype B. The infection of genotype D and mixed infection also occurred in the Li nationality. Genotype C HBV has a higher replication rate, and the different degrees of pathogenecity among HBV genotypes may be related to BCP T1762/A1764 mutation frequency.

Biliary complications(BC) are a major cause of morbidity in liver transplant recipients with an incidence of 10∼30% following orthotopic liver transplantation(OLT), and a mortality rate of up to 10%. The most common biliary complications are bile leaks, biliary strictures, ampullary dysfunction, and stones. Leaks predominate in the early posttransplant period; while stricture formation typically develops gradually over time. Risk factors for biliary complications comprise technical failure, T-tube-related complications, hepatic artery thrombosis, bleeding, ischemia/reperfusion injury, primary diseases, and other immunological, non-immunological, and infectious complications. Cholangiography, such as endoscopic retrograde cholangiopancreatography(ERCP) or percutaneous transhepatic cholangiogram(PTC), is considered the gold standard for identifying post-transplant BC. The management of biliary complications after OLT requires a multidisciplinary approach, in which interventional radiology and endoscopic techniques are emerging as the preferred treatment option, but in a selected majority of patients, surgery is still necessary.

A 67-year-old woman with chronic cholecystitis was scheduled to have laparoscopic cholecystectomy under general anesthesia. About 5∼10 min after the CO₂ intraperitoneal insufflation, the peak airway pressure gradually increased from 15 cmH₂O to 27 cmH₂O, the end-tidal CO₂(EtCO₂) from 32 mmHg to 56 mmHg. The SpO₂ decreased from 100% to 96%, and blood pressure from 135/80 mmHg to 80/52 mmHg. A right side tension pneumothorax was confirmed and a drainage tube was placed in the right pleural cavity. As the continuous gas leakage from the drainage tube was noted, even as ventilation was withheld, the diaphragm was carefully examined and a porous diaphragm was found. These defects were then patched with biomedical materials. The operation was finished uneventfully. It was concluded that in a patient with a tension pneumothorax during laparoscopic surgery, a diaphragm defect should be taken into consideration.

Objective

To explore the antibacterial effect of Chinese crude drugs against clinical strains of Ureaplasma urealyticum(UU), including eight pure herbs and three compound herbs, and determine their minimal inhibitory concentrations(MICs).

Methods

Isolates were collected from clinical patients with UU infection, and cultured in UU broth. In order to test the different effects on clinical strains of UU, the assays were performed by microdilution inhibition tests, and MICs of the herbs against the clinical strains of UU were calculated.

Results

The MICs of eight pure herbs against clinical strains of UU were as follows: Galla Chinensis:0.313∼1.25 g/L; Fructus Forsythiae: 1.25∼5.00 g/L; Cortex Phellodendri:1.25∼5.00 g/L; Radix Paeoniae Rubra:1.25∼2.50 g/L; Semen Plantaginis:2.50∼10.00 g/L; Herba Lysimachiae: 5.00∼20.00 g/L; Rhizoma Coptidis: 5.00∼20.00g/L, Herba Houttuyniae: 10.00∼20.00 g/L. The MICs of compound herbs were: Liuheji: 0.625∼2.50 g/L; Bazhengsan: 5.00∼20.00 g/L; Wulinsan:2.50∼20.0 g/L.

Conclusion

Galla Chinensis, Fructus Forsythiae, Cortex Phellodendrim, Radix Paeoniae Rubra, and Semen Plantaginis, exerted the stronger antibacterial effect against clinical strains of UU, whereas Herba Lysimachiae, Rhizoma Coptidis and Herba Houttuyniae, had relatively weaker activity against UU. Compound herbs, Bazhengsan and Wulinsan, and particularly Liuheji, also had antibacterial effects against UU. Further studies of the effects and mechanisms of action of Chinese crude drugs against UU infections are worthwhile.

Objective

To investigate the effects of siRNA of ADAM17 gene and genistein on apoptosis and the inhibition of proliferation in HepG2 cells in an attempt to seek an effective therapy for hepatocellular carinoma.

Methods

Cells were divided into control groups and experimental groups and siRNA was used to silence the ADAM17 gene, alone and in combination with genistein. Cells were harvested at several time periods and assessed for proliferation and apoptosis. Proliferation was assayed by MTT at 24, 48, 72 and 96 hours following treatment and apoptosis was assessed by flow cytometric analysis at 48 hours.

Results

In siRNA groups, proliferation of cells was significantly inhibited compared to the control groups at 24, 48 and 72 hours(P < 0.05), and apoptosis was significantly increased at 48 hours(P < 0.01); In genistein groups, proliferation was inhibited at 24, 48, 72 and 96 hours, and the apoptosis ratio was significantly increased at 48 hours(P < 0.01); while in the groups that received the combination of siRNA transfection and genistein treatment, there was a further significant decrease of proliferation and increase in apoptosis compared with either treatment alone.

Conclusion

The ADAM17 gene could be an effective target, and genistein could be a useful agent, in the treatment of hepatocellular carcinoma. siRNA of ADAM17 gene and genistein both inhibited HepG2 cells proliferation and promoted apoptosis, and further, the combination of these treatments had a greater effect than either treatment alone.