The first juvenile specimen of Manchurochelys manchoukuoensis from the Early Cretaceous Jehol Biota.

A small juvenile turtle is described from the Early Cretaceous Jehol Biota, shedding light on the juvenile morphology and ontogeny of Manchurochelys manchoukuoensis. Several juvenile features are uncovered, such as a small and circular carapace (less than half of the adult), wide vertebral scales, and lateral carapacial fontanelles. In contrast to the adult morphology, which has an oval carapace, closed lateral fontanelles, and longer vertebrals 2-4, the juvenile of M. manchoukuoensis is more comparable to that of Sinemys lens, except for earlier occurrence of the well-ossified carapace of the latter. Differs from Changmachelys bohlini, and Ordosemys liaoxiensis, in which the circular carapace is relatively independent of ontogenetic age, and the lateral fontanelles are only closed in adult stage of O. liaoxiensis. Therefore, the trajectory of ontogenetic change appears to be highly diversified in the sinemydids.

Analysis of the prognostic factors for low rectal cancer with the pT1-2NxM0 stage after abdominoperineal resection.

OBJECTIVE: This study was designed to explore the factors influencing local recurrence and survival for low rectal cancer with pT1-2NxM0 stage after an abdominoperineal resection (APR). METHODS: Data of 429 patients confirmed to have pT1-2NxM0 after APR were reviewed. RESULTS: The recurrence rate in patients with intraoperative perforation, less than 12 lymph nodes (LNs) harvested, T2 staging, and positive circumferential resection margin (CRM) was 25.1, 19.9, 9.5, and 26.1% compared with 6.9, 7.0, 0, and 5.8% in patients with no perforation, 12 or more LNs harvested, T1, and negative CRM. The 5-year survival rate in patients with age of at least 70, perforation, less than 12 LNs harvested, T2, and positive CRM was 71.1, 60.8, 58.8, 69.9, and 46.0%, but 73.4, 73.5, 73.8, 89.4, and 75.0% in patients with age less than 70, no perforation, 12 or more LNs harvested, T1, and negative CRM. Meanwhile, patients with N0, N1, and N2 had a survival rate of 90.7, 69.9, and 63.9%. Multivariate analysis showed that perforation (P<0.001), number of LNs harvested (P<0.001), T staging (P<0.001), differentiation (P=0.045), and CRM status (P=0.002) were associated with local recurrence, whereas age of the patients (P=0.023), N staging (P<0.001), differentiation (P=0.011), and CRM status (P=0.004) were associated with survival. CONCLUSION: APR was affected by patients' age, operation performer, perforation, number of LNs harvested, T staging, N staging, differentiation, and CRM status. Perforation, number of LNs harvested, T staging, differentiation, and CRM status were independent factors for recurrence; meanwhile, age of the patients, N staging, differentiation, and CRM status were independent factors influencing survival.

miR-449a and CDK6 in gastric carcinoma.

The present study aimed to identify the association between microRNA (miR/miRNA)-449a, the cyclin-dependent kinase (CDK)6 protein and gastric carcinoma, and discuss the effect of miR-449a on the expression of the CDK6 protein. Quantitative (q)PCR and western blot analysis were used to analyze the expression of the miR-449a and the CDK6 protein in gastric carcinoma and tumor-adjacent normal tissues. The real-time cell analyzer and the DAPI staining test were used to monitor the different miR-449a levels regulating the proliferation and apoptosis of the MGC-803 cell line. Immunofluorescence and western blot analyses were used to detect the expression level of the CDK6 protein in the cells of the miR-449a upregulation and downregulation groups, and a control group. A scratch test was used to study the effects of miR-449a expression on migration and invasion. It was found that the expression of miR-449a was downregulated and the expression of CDK6 protein was upregulated in gastric carcinoma tissue. The level of MGC-803 cell proliferation was decreased and the apoptosis level was increased by the upregulation of miR-449a expression, and the opposite effect was shown by the downregulation of expression. The expression of the CDK6 protein in the MGC-803 cells was downregulated by upregulating the expression of miR-449a. The distance of the scratch was shortened markedly after 12 h by downregulating the expression of miR-449a in the MGC-803 cells. The present study identified that a lower expression level of miR-449 and a higher expression level of CDK6 may contribute to the occurrence and development of gastric cancer. Furthermore, it was shown that miR-449a is able to regulate the expression of the CDK6 protein.

Undifferentiated embryonal sarcoma of the liver mistaken for hepatic abscess in an adult.

Undifferentiated embryonal sarcoma of the liver (UESL) predominantly occurs in children under the age of 10 years, and ~90% of cases occur in children <15 years old. Patients may complain of abdominal pain, fever or other symptoms. No significant decrease has been identified in the hepatic function or elevation of α-fetoprotein, which differentiates UESL from primary carcinomas of the liver. In the present study, a rare and misdiagnosed case of an UESL arising in a male, which was mistaken for a hepatic abscess and retrospectively re-diagnosed, is reported. This case was misdiagnosed as a hepatic abscess initially, and it was diagnosed as UESL subsequent to performing tests, including a type-B ultrasonic scan and computed tomography (CT), and evaluating pathological findings. The rapid recurrence of the tumor in this patient was identified by CT, and this is associated with the malignancy of the disease. Currently, patients with UESL have a poor prognosis as there is not a successful treatment strategy. The present study analyzes the course of diagnosis and potential treatment for the disease.

The protective effect of recombinant Lactococcus lactis oral vaccine on a Clostridium difficile-infected animal model.

BACKGROUND: Oral immunization with vaccines may be an effective strategy for prevention of Clostridium difficile infection (CDI). However, application of previously developed vaccines for preventing CDI has been limited due to various reasons. Here, we developed a recombinant Lactococcus lactis oral vaccine and evaluated its effect on a C. difficile-infected animal model established in golden hamsters in attempt to provide an alternative strategy for CDI prevention. METHODS: Recombinant L. lactis vaccine was developed using the pTRKH2 plasmid, a high-copy-number Escherichia coli-L. shuttle vector: 1) L. lactis expressing secreted proteins was constructed with recombinant pTRKH2 (secreted-protein plasmid) carrying the Usp45 signal peptide (SPUsp45), nontoxic adjuvanted tetanus toxin fragment C (TETC), and 14 of the 38 C-terminal repeats (14CDTA) of nontoxic C. difficile toxin A (TcdA); and 2) L. lactis expressing secreted and membrane proteins was constructed with recombinant pTRKH2 (membrane-anchored plasmid) carrying SPUsp45, TETC, 14CDTA, and the cell wall-anchored sequence of protein M6 (cwaM6). Then, 32 male Syrian golden hamsters were randomly divided into 4 groups (n = 8 each) for gavage of normal saline (blank control) and L. lactis carrying the empty shuttle vector, secreted-protein plasmid, and membrane-anchored plasmid, respectively. After 1-week gavage of clindamycin, the animals were administered with C. difficile spore suspension. General symptoms and intestinal pathological changes of the animals were examined by naked eye and microscopy, respectively. Protein levels of anti-TcdA IgG/IgA antibodies in intestinal tissue and fluid were analyzed by enzyme-linked immunosorbent assay (ELISA). A cell culture cytotoxicity neutralization assay was done by TcdA treatment with or without anti-TcdA serum pre-incubation or treatment. Apoptosis of intestinal epithelial cells was examined by flow cytometry (FL) assay. Expression of mucosal inflammatory cytokines in the animals was detected by polymer chain reaction (PCR) assay. RESULTS: After the C. difficile challenge, the animals of control group had severe diarrhea symptoms on day 1 and all died on day 4, indicating that the CDI animal model was established in hamster. Of the 3 immunization groups, secreted-protein and membrane-anchored plasmid groups had significantly lower mortalities, body weight decreases, and pathological scores, with higher survival rate/time than the empty plasmid group (P < 0.05). The tilter of IgG antibody directed against TcdA was significantly higher in serum and intestinal fluid of secreted-protein and membrane-anchored plasmid groups than in the empty plasmid group (P < 0.05) while the corresponding titer of IgA antibody directed against TcdA had no substantial differences (P > 0.05). The anti-TcdA serum of membrane-anchored plasmid group neutralized the cytotoxicity of 200 ng/ml TcdA with the best protective effect achieved by anti-TcdA serum pre-incubation. The incidences of TcdA-induced death and apoptosis of intestinal epithelial cells were significantly reduced by cell pre-incubation or treatment with anti-TcdA serum of membrane-anchored plasmid group (P < 0.05). MCP-1, ICAM-1, IL-6, and Gro-1 mRNA expression levels were the lowest in cecum tissue of the membrane-anchored groups compared to the other groups. CONCLUSION: Recombinant L. lactis live vaccine is effective for preventing CDI in the hamster model, thus providing an alternative for immunization of C. difficile-associated diseases.

Changes in innate and permissive immune responses after hbv transgenic mouse vaccination and long-term-siRNA treatment [corrected].

BACKGROUND: Currently, no licensed therapy can thoroughly eradicate hepatitis B virus (HBV) from the body, including interferon α and inhibitors of HBV reverse-transcription. Small interfering RNA (siRNA) seem to be a promising tool for treating HBV, but had no effect on the pre-existing HBV covalently closed circular DNA. Because it is very difficult to thoroughly eradicate HBV with unique siRNAs, upgrading the immune response is the best method for fighting HBV infection. Here, we aim to explore the immune response of transgenic mice to HBV vaccination after long-term treatment with siRNAs and develop a therapeutic approach that combines siRNAs with immunopotentiators. METHODOLOGY/PRINCIPAL FINDINGS: To explore the response of transgenic mice to hepatitis B vaccine, innate and acquired immunity were detected after long-term treatment with siRNAs and vaccination. Antiviral cytokines and level of anti-hepatitis B surface antigen antibody (HBsAg-Ab) were measured after three injections of hepatitis B vaccine. RESULTS: Functional analyses indicated that toll-like receptor-mediated innate immune responses were reinforced, and antiviral cytokines were significantly increased, especially in the pSilencer4.1/HBV groups. Analysis of CD80+/CD86+ dendritic cells in the mouse liver indicated that dendritic cell antigen presentation was strengthened. Furthermore, the siRNA-treated transgenic mice could produce detectable HBsAg-Ab after vaccination, especially in the CpG oligonucleotide vaccine group. CONCLUSIONS/SIGNIFICANCE: For the first time, our studies demonstrate that siRNAs with CpG HBV vaccine could strengthen the immune response and break the immune tolerance status of transgenic mice to HBV. Thus, siRNAs and HBV vaccine could provide a sharp double-edged sword against chronic HBV infection.

[Endoscopic nasobiliary drainage for acute obstructive suppurative cholangitis with multiple organ failure: report of 25 cases].

OBJECTIVE: To explore the treatment of acute obstructive suppurative cholangitis (AOSC) with multiple organ failure (MOF). METHODS: Twenty-five patients with AOSC complicated by MOF underwent non-surgical comprehensive therapies, including endoscopic naso biliary drainage (ENBD), flushing and antibiotic perfusion through the naso biliary catheter. The alterations of the levels of serum total bilirubin and common bile duct diameter were measured both preoperatively and postoperatively, with retrospective analysis of the patients' clinical record. RESULTS: Of all the 25 patients, 23 underwent endoscopic retrograde cholangiopancreatography (ERCP) with successful placement of the nasobiliary catheters and bile drainage, and MOF was corrected. The cure rate of the this group of patients was 92.0%, with two cases being transferred for emergency surgical treatment. CONCLUSION: Comprehensive treatment consisting of ENBD, flushing and antibiotic perfusion through the naso biliary catheter and intravenous use of antibiotics is effective and safe for the treatment of AOSC complicated by MOF.

Experimental study on mechanism and protection of stress ulcer produced by explosive noise.

AIM:To establish an experimental model of stress ulcer produced by explosive noise, and to probe into its mechanism and protection.METHODS:The country standard Wistar white rats were randomly divided into control group(n =8), which were neither stimulated nor protected, and stimulating group (divided into subgroups A, B and C, including 8 rats each which were decapitated to draw blood for test immediately, 12 hours and 24 hours after stimulation) and prevention group (divided into subgroups A, B and C, having 8 rats each, subgroup A was given cimetidine, B anisodamine and C both drugs). Firing noises of submachine guns were used as inflicting factor. The rats were fasted for 24 hours and stimulated by firing noise for 12 hours. The change of ulcer index, gastric mucosal and related serum hormones were observed.RESULTS:Stress ulcer was significant in the stimulating group, and its ulcer index (8.6 ± 0.6) was remarkably higher than that in both the control group and prevention group (0.3 ± 0.1, P < 0.01).Its serum gastrin (Gas ng/L, 294 ± 163 vs 63 ± 40,P< 0.01) and endothelin (ET ng/L, 181 ± 57 vs 135 ± 42, P < 0.1) were apparently higher than those in the control group, and its serum nitric oxide (NO) level was conspicuously lower than that in the control group (ng/L, 0.3 ± 0.1 vs 0.8 ± 0.5 P <0.5), while the serum gastrin level (ng/L, 556 ± 225) in prevention group was distinctly higher than that in both the control (P<0.1) and stimulating group (P < 0.5). There were no significant differences in the changes of ET and NO between the control and the stimulating groups.CONCLUSION:Stress ulcer model of rats can be successfully established by the stimulation of explosive noise. Gas, ET and NO are related to the formation of stress ulcer, and play an important role in its mechanism. Hepatic function affected by noise is observed in this experiment.