Objective To evaluate the prognostic significance of p53, p21/WAF1 and pRb in T 1G 3 transitional cell carcinoma (TCC) of the urinary bladder. Methods Paraffin embedded sections from 47 patients with T 1G 3 TCC of the bladder were studied by immunohistochemistry, using monoclonal antibodies of p53, p21/WAF1 and pRb. The immunostaining results were analyzed in relation to the tumor progression, and compared with the clinical parameters on prognostic significance. Results Of the 39 patients treated by TURBT or partial cystectomy, the total progression rate of the tumor has been 59%.Of the 8 patients treated by cystectomy,one had pulmonary metastasis 2 years after the operation.The abnormal expression rates of p53, p21/WAF1 and pRb were 66.7%, 51.4% and 71.8% respectively. Multivariate analysis indicated that only the combined abnormal expressions of p53, p21/WAF1 and pRb provided prognostic information in T 1G 3 TCC of the bladder. Conclusions Immunohistochemical evaluation of p53, p21/WAF1 and pRb nuclear expressions would help the prognostic evaluation cases of T 1G 3 TCC of the bladder.

OBJECTIVETo investigate the effect of Zilongjin (ZLJ) on human androgen-dependent type of prostate cancer cell line LNCaP.

METHODSMTT assay, flow cytometry and fluorescence microscopy were used to observe the effect of ZLJ in anti-proliferation, cell cycle arresting and apoptosis induction. RT-PCR was used to examine the effect of ZLJ on expressions of prostate marker gene (PSA), androgen receptor (AR), apoptosis related genes (bcl-2 and bax), and Western blot assay was used to detect the effect on protein expression of bcl-2 and bax.

RESULTSZLJ could cause apparent inhibition on proliferation, induce G0/G1 phase arresting and apoptosis in time- and dose-dependent manner on LNCaP cells. The concentration for inhibiting cell growth by 50% (IC50) in 72 hrs was 0.79 mg/ml. ZLJ could down-regulate the expression of PSA, AR, bcl-2 genes and lower bcl-2 protein expression, but showed ineffective on bax protein expression.

CONCLUSIONZLJ displays its anti-tumor effects by way of inhibiting the cell proliferation, arresting the G0/G1 phase, inducing apoptosis, down-regulating PSA, AR, bcl-2 gene expression and lowering bcl-2 protein expressions.

Antineoplastic Agents, Phytogenic ; pharmacology ; Cell Line, Tumor ; Cell Proliferation ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Humans ; Male ; Neoplasms, Hormone-Dependent ; metabolism ; pathology ; Prostate-Specific Antigen ; biosynthesis ; genetics ; Prostatic Neoplasms ; metabolism ; pathology ; Proto-Oncogene Proteins c-bcl-2 ; biosynthesis ; genetics ; Receptors, Androgen ; biosynthesis ; genetics

Objective: To observe the effect of dopamine pretreatment of the root canal on improving the bonding performance of AH-plus sealer. Methods : A total of 32 freshly isolated permanent teeth with a single canal were collected, with no caries, no fracture of roots, and a root canal curvature<10°. All sample root canals were prepared to F2 with ProTaper rotating nickel-titanium instruments and then treated with 1 mg/mL, 2 mg/mL, or 3 mg/mL dopamine solution for 24 hours and divided into 4 groups (n = 8): 0 mg/mL dopamine group (blank control group), 1 mg/mL dopamine group, 2 mg/mL dopamine group, and 3 mg/mL dopamine group. Scanning electron microscopy was used to observe the combination of dopamine and root canal dentin wall; laser confocal scanning microscopy was used to observe the penetration of AH-plus sealer; and root canal filling was performed with AH-plus sealer and gutta-percha tip using the cold gutta-percha lateral pressure technique. The root canal samples were cut horizontally at the middle and the apical third sections of the root with a slice thickness of 1-2 mm. The push-out test was carried out under an Instron universal testing machine to compare the push-out bonding strength between each group. Results : Scanning electron microscopy showed that most of the dentinal tubules were open in the control group after 0 mg/mL dopamine solution treatment for 24 hours. In the 1 mg/mL group, a small number of dopamine particles on the surface of the dentin tubules in the inner wall of the root canal were loose and unevenly distributed. In the 2 mg/mL group, most of the dentinal tubules were covered by dopamine particles, and the dopamine layer was uniform and dense. In the 3 mg/mL group, a large number of dopamine particles were deposited at the mouth of the dentinal tubules, but the distribution was uneven. Dopamine and AH-plus sealer can be seen to simultaneously infiltrate into dentinal tubules under a confocal laser scanning microscope. The interaction of the two factors, the anatomical location and dopamine concentration, had no significant effects on the bonding strength of AH-plus sealer (P>0.05). Root canals treated with 2 mg/mL dopamine had the highest bonding strength in all groups (P<0.05). Analysis of the push-out test of bonding strength with AH-plus sealer at different anatomical locations showed significant differences (P<0.05). The push-out bonding strength of the AH-plus sealer in the middle third section of the root was higher than that in the apical third section of the root. Conclusion Different dopamine concentrations could affect the bonding strength of AH-plus sealer in root canals. When treated with 2 mg/mL dopamine for 24 hours, the bonding effect of AH-plus sealer in root canals was improved.

Auditory neuropathy spectrum disorder (ANSD) represents a variety of sensorineural deafness conditions characterized by abnormal inner hair cells and/or auditory nerve function, but with the preservation of outer hair cell function. ANSD represents up to 15% of individuals with hearing impairments. Through mutation screening, bioinformatic analysis and expression studies, we have previously identified several apoptosis-inducing factor (AIF) mitochondria-associated 1 (AIFM1) variants in ANSD families and in some other sporadic cases. Here, to elucidate the pathogenic mechanisms underlying each AIFM1 variant, we generated AIF-null cells using the clustered regularly interspersed short palindromic repeats (CRISPR)/CRISPR-associated protein 9 (Cas9) system and constructed AIF-wild type (WT) and AIF-mutant (mut) (p.‍T260A, p.‍R422W, and p.‍R451Q) stable transfection cell lines. We then analyzed AIF structure, coenzyme-binding affinity, apoptosis, and other aspects. Results revealed that these variants resulted in impaired dimerization, compromising AIF function. The reduction reaction of AIF variants had proceeded slower than that of AIF-WT. The average levels of AIF dimerization in AIF variant cells were only 34.5%‍‒‍49.7% of that of AIF-WT cells, resulting in caspase-independent apoptosis. The average percentage of apoptotic cells in the variants was 12.3%‍‒‍17.9%, which was significantly higher than that (6.9%‍‒‍7.4%) in controls. However, nicotinamide adenine dinucleotide (NADH) treatment promoted the reduction of apoptosis by rescuing AIF dimerization in AIF variant cells. Our findings show that the impairment of AIF dimerization by AIFM1 variants causes apoptosis contributing to ANSD, and introduce NADH as a potential drug for ANSD treatment. Our results help elucidate the mechanisms of ANSD and may lead to the provision of novel therapies.
Humans ; Apoptosis Inducing Factor/metabolism* ; NAD/metabolism* ; Dimerization ; Apoptosis