Carvedilol alleviates the biliary cirrhosis through inhibiting the endoplasmic reticulum stress.

OBJECTIVE: To investigate the effects of carvedilol on inflammation, apoptosis, and hepatic fibrosis caused by biliary cirrhosis and its mechanisms in mice. MATERIALS AND METHODS: 60 male C57/BL6 mice were randomly divided into sham-operation group (Sham group, n=20), biliary cirrhosis group (BDL group, n=20) and carvedilol group (CAR group, n=20). The CAR group was treated with gavage using 12.5 mg/kg carvedilol, once a day for 14 consecutive days, while the Sham group and BDL group were treated with gavage using the equivalent normal saline. After that, the mice in Sham group received the laparotomy under chloral hydrate anesthesia, followed by direct abdominal closure. The mice in BDL group and CAR group received the common bile duct ligation after anesthesia for modeling. After modeling, the survival rate of mice in each group was detected, and the blood and liver tissues were taken for detection. The morphological changes in liver tissues and apoptosis in mice in each group were detected and compared. The levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), total bilirubin (TBIL), superoxide dismutase (SOD), malondialdehyde (MDA), hydroxyproline, and α-smooth muscle actin (α-SMA) were also detected. The mRNA expression levels of pro-inflammatory factors, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), transforming growth factor ß-1 (TGF-ß1), α-SMA and collagen-1 were detected via reverse transcription-polymerase chain reaction (RT-PCR). The protein expression levels of CHOP (CCAAT-enhancer-binding protein homologous protein), activating transcription factor 4 (ATF4), ATF6, inositol-requiring enzyme 1 (IRE1), phosphorylated Jun N-terminal kinase (pJNK), α-SMA, and collagen-1, were detected via Western-blotting. RESULTS: Our study showed that carvedilol could significantly alleviate the biliary cirrhosis in mice, and improve the survival rate of mice. The ALT, AST and TBIL levels, severity of cirrhosis, and number of apoptotic cells in CAR group were significantly lower than those in BDL group. The levels of α-SMA and hydroxyproline in CAR group were also significantly lower than those in BDL group. The activity of SOD in CAR group was significantly higher than that in BDL group; the above differences were statistically significant (p<0.05). In addition, it was also found that carvedilol could down-regulate the mRNA expression levels of iNOS, COX-2 and TGF-ß1, down-regulate the mRNA and protein expression levels of α-SMA and collagen-1, and negatively regulate the ATF4-CHOP, ATF6-CHOP and IRE1-pJNK signaling pathways. CONCLUSIONS: Carvedilol has a significant effect on alleviating the biliary cirrhosis in mice, and its relevant mechanism may be that carvedilol inhibits the endoplasmic reticulum stress through the negative regulation of ATF4-CHOP, ATF6-CHOP and IRE1-pJNK signaling pathways, which needs to be confirmed by further in vitro experiments.

[Diagnostic value of narrow-band imaging in detection of recurrent nasopharyngeal carcinoma].

Objective: To evaluate the diagnostic value and feasibility of narrow-band imaging in detection of recurrent nasopharyngeal carcinoma (NPC). Methods: One thousand three hundred and sixty-four NPC patients who had completed NPC treatment were enrolled. All patients were followed-up with imaging, serological examination of EB virus and nasopharyngeal endoscopy(WL and NBI mode), in which (1) both white light (WL) and NBI modes were done; (2) positive endoscopic patients were given nasopharyngeal biopsy; (3) using histologic finding as criterion standard, the sensitivity, specificity, accuracy and Yonden's index of two modes were compared. Kappa index was used to evaluate the consistency between the two modes and pathological results respectively; (4) the positive rates of WL and NBI in patients with early recurrent (stage Ⅰ+ Ⅱ) were compared. Results: A total of 265 cases were suspected as having recurrent lesions by endoscopy in WL mode and 68 cases of them were pathologically diagnosed as having NPC; and 82 cases were suspected as having recurrent lesions by endoscopy in NBI mode and 74 cases of them were pathologically diagnosed as having NPC. The sensitivity, specificity, accuracy and Yonden's index of WL mode were 91.89%, 0, 25.09% and -0.0811, respectively, with a kappa of -0.045; the sensitivity, specificity, accuracy and Yonden's index of NBI mode were 100.00%, 95.94%, 97.05% and 0.9594, respectively. Conclusion: NBI has higher sensitivity, specificity, early diagnosis rate and Yonden's index than WL.

A research for the relationship between human papillomavirus and human uterine cervical carcinoma. I. The identification of viral genome and subgenomic sequences in biopsies of Chinese patients.

Biopsies from 318 cases with squamous cell carcinoma of the uterine cervix, 48 with cervical and vulvar condylomata, 14 with cervical intraepithelial neoplasia (CIN), 34 with chronic cervicitis and 24 with normal cervical epithelium were collected from different geographic regions with different cervical cancer mortalities. The DNA.DNA dot-blot and Southern blot hybridization results show that there is a close relationship between HPV-16 and the uterine cervical squamous cell carcinoma in China. One very interesting observation is that the finding of HPV-16-homologous DNA differs significantly among five geographic regions, and corresponds with the mortalities from cervical cancer of these five regions. HPV-11 was found mainly in benign lesions. The rate of detection of HPV-16 in Chinese women increased from 8.3% in normal cervical epithelium to 20% in chronic cervicitis, 28% in cervical condyloma, 50% in CIN and 60.4% in cervical cancer. It is suggested that HPV-16 infection may be an etiological factor in the development of human cervical carcinoma. From the results of Southern blot hybridization, it appeared that HPV-16 DNA had been integrated into the genome of the host cell in cervical cancer. Whereas the HPV-16 DNA sequence was only present as an episome in normal cervical epithelium and cervical benign lesions. The rate of occurrence of E6-E7 genes is the highest (88.9%) compared with that of other subgenomic fragments of HPV-16 in specimens of human cervical cancer in China. This implies that E6 and E7 may be the oncogenic genes of HPV-16 and play an important role in the carcinogenesis of human cervical epithelial cells. The amplification and rearrangement of the c-myc protooncogene are closely associated with the occurrence of cervical cancer. The results presented here revealed that the activated c-myc oncogene may cooperate with HPV-16 in the carcinogenic processes.

A research for the relationship between human papillomavirus and human uterine cervical carcinoma. II. Molecular genetic and ultrastructural study on the transforming activity of recombinant retrovirus containing human papillomavirus type 16 subgenomic sequences.

In order to elucidate the role of HPV-16 in the development of genital cancer, NIH3T3 cells were transfected by HPV-16 whole genome and its two early genes, E6-E7. Besides ordinary calcium phosphate/DNA coprecipitation technique, a newly designed recombinant retrovirus containing the HPV-16 genome or subgenomes was used to infect cells for transfer of the target genes. The transforming activities have been demonstrated to be most efficient when a bioengineering technique of this kind is used. HPV-16 DNA was proved to have transforming potential for NIH3T3 cells, and the DNA of HPV-16 was proved to undergo multisite integration into transformed cells and nude mice tumour cells. The E6-E7 open reading frames are sufficient for transforming NIH3T3 cells independently in vitro, which implies that E6-E7 open reading frames are transforming genes or even viral oncogenes of HPV-16. The RNA transcribed by the E6-E7 of HPV-16 was expressed in transformed cells and in tumour cells of nude mice. The use of a recombinant retrovirus for gene transfer in this study is much more efficient than that of calcium phosphate/DNA coprecipitation. The lack of a tissue-culture system suitable for HPV replication in vitro makes HPV gene recombination into a specially engineered retrovirus for viral-mediated gene transfer of particular significance for the possible application of viral carcinogenesis, both in vitro and in vivo, for basic and clinical research.

Histopathological and ultrastructural changes of lichen sclerosus et atrophicus on the vulva.

Studies of 82 skin specimens from 81 cases of lichen sclerosus et atrophicus (LSA) are reported. Typical LSA pathological changes showed atrophy of the epidermis, hydropic degeneration at the basal layer, and homogenization of the collagen fibers in the superficial dermis with inflammatory cell infiltration underneath. Ultrastructural changes showed intercellular spaces enlarged, particularly near basal cells. Some mitochondria were swollen and rough endoplasmic reticula were enlarged. Some basement membranes were thinner, collagen fibers of varying diameters were arranged in irregular shapes and the fine structures had become unclear and had sometimes even disappeared.

Viral etiology of cervical carcinoma. Human papilloma virus and herpes simplex virus type 2.

The possible role of human papilloma virus (HPV) and herpes simplex virus type 2 (HSV-2) in the viral etiology of cervical carcinoma was investigated a series of cervical lesions were studied for the presence of HPV and HSV-2 DNA sequences as well as HPV and HSV-2 antigens by DNA dot blot hybridization technique and high-specificity PAP staining method. The results obtained were correlated with the histologic diagnosis. HPV 16 DNA sequences detected in cervical carcinoma biopsies were 43%, whereas HSV-2 DNA sequences were only 8%. HPV antigens detected in cervical dysplasia were 31%, whereas those detected in cervical carcinoma and cervicitis were the least. HSV-2 antigens were detected in chronic cervicitis, dysplasia and cervical carcinoma. The difference in positive rate between the cervical carcinoma and cervicitis groups was statistically significant, (chi-square test, P less than 0.01). No HPV DNA and HSV-2 DNA sequences were found in the same specimen, although both HPV DNA sequences and HSV-2 antigens were found in the same sample in some cases. The results indicate that the viral etiology of cervical carcinoma may be multifactorial. Both HSV-2 and HPV may be associated with cervical carcinoma, but the mechanisms involved are different. HSV-2 and HPV may act synergistically in the development of cervical carcinoma.

[Analysis of the characteristics of human papilloma virus infection in 85 neoplasms of the respiratory system in adult patients].

Eighty five neoplastic tissues of respiratory system in Chinese adult patients were searched for HPV by multiple PCR and in situ hybridization. In 13 of the specimens examined, HPV DNA was identified (15.3%), including 6 of 36 cases of pharyngolaryngeal neoplasms (16.6%) and 7 of 49 cases of lung squamous cell carcinomas (14.2%). Among the 13 cases tumor specimen, most of them were positive for HPV6/11 types and a few were double positive for HPV 6/11 and 16 types. Many tumor cells positive for HPV DNA on in situ hybridization were morphologically similar to those koilocytes in genital condylomas. HPV infection in 85 neoplasms was characterized by the fact that the same genotypes of HPV were detected in upper and lower respiratory tract and that the dominant virus types detected were HPV 6/11 types. The detection rate of HPV was a little higher in the upper than that in the lower respiratory tract and the HPV DNA positive tumors had similar histological appearance. The results suggest that HPV infection in the neoplasms of the upper and the lower airways is related to each other with similar mode of transmission.

[Use of polymerase chain reaction with L1 consensus primer for detection of HPV16 and HPV18 in cervical cancer tissues].

Biopsy materials of cervical carcinoma including 20 cervical adenocarcinomas and 20 squamous cell carcinomas were collected. A rapid method for determining HPV type was developed, based on DdeI restriction enzymes analysis within the L1 region of HPV, amplified by PCR using consensus primers. The results indicated that HPV type 16 was detected more often in squamous cell carcinomas than in adenocarcinomas (80% vs 15%, P < 0.001), conversely, HPV type 18 was detected significantly more often in adenocarcinoma tissues (45% vs 5%, P < 0.001). These differences may reflect the presence of different virus receptors in cancer cells with different morphologic potential, or, they may indicate that the specific HPV infection actually plays a direct role in the process of carcinogenesis.

[A study of human papillary virus infection by in situ hybridization and histopathology in squamous cell carcinoma of the lung].

Paraffin embedded archival tissue from 7/49 carcinomas of the lung which were positive for HPV DNA examined by multiple PCR were again searched for HPV DNA by in situ hybridization technique. Five cases were found positive for HPV DNA by in situ hybridization, of which three cases were positive for HPV 11, One was positive for HPV 16 and the other was positive for both HPV 11 and 16. The hybridization signal was mainly located in the nuclei of the koilocytotic tumor cells. These results suggest that HPV infection may have some role in the development of squamous cell carcinoma of the lung.

[Detection of human papillomavirus DNA in squamous cell carcinomas of the lung by multiple polymerase chain reaction].

49 cases of formalin-fixed paraffin-embedded tissues of squamous cell carcinomas of the lung were searched for the presence of human papillomavirus (HPV) DNA by multiple polymerase chain reaction (PCR). Totally seven cases were positive for HPV DNA (14.2%), Among them four cases were positive for HPV6/11, one case was positive for HPV16 and two other cases were double positive for HPV6/11 and 16. These results suggest that HPV infection might play some role in development of squamous cell carcinomas of the lung.

Cimifoetisides VI and VII Two new cyclolanostanol triterpene glycosides from the aerial parts of Cimicifuga foetida.

Two new cyclolanostanol triterpene glycosides, cimifoetiside VI (1) and cimifoetiside VII (2), and one known compound were isolated from the aerial parts of Cimicifuga foetida L. On the basis of spectral and chemical evidences, the structures of 1 and 2 were elucidated to be (23R,24S)-24-O-acetylisodahurinol-3-O-beta-d-galactopyranoside and (23R,24R)-24-O-acetylshengmanol-3-O-beta-d-glucopyranosyl-(1'' --> 2')-beta-d-xylopyranoside. The known compound was identified as (23R,24R)-24-O-acetylshengmanol-3-O-beta-d-galactopyranoside (3).

Chemical constituents of Typhonium giganteum Engl.

A new cerebroside, named typhonoside (1), was isolated from the root tuber of Typhonium giganteum Engl. along with three known compounds dipalmitin (2), alpha-monopalmitin (3) and 2,6-diamino-9-beta-D-ribofuranosylpurine (4). The structure of 1 was determined to be 1-O-beta-D-glucopyranosyl-(2S,3S,4R,8Z)-2-[(2'-hydroxyl-docosanoyl)amino]-8-otadecene-1,3,4-triol on the basis of spectral data.