[The expressions of CTLA-4 and PD-1 on CD(4)(+) T cells and the level of plasma VEGF in patients with obstructive sleep apnea hypopnea syndrome].

Objective: CD(4)(+)T cells, cytotoxic T-lymphocyte antigen 4 (CTLA-4), programmed cell death-1 (PD-1) and vascular endothelial growth factor (VEGF) are associated with cancer development. The aim of the present study was to investigate the expression of CTLA-4, PD-1 and VEGF in patients with obstructive sleep apnea hypopnea syndrome (OSAHS). Methods: From January 2017 to January 2018, a total of 47 first-visit outpatients were recruited in the Sleep and Respiratory Disorder Center of Guangdong Provincial People's Hospital, and were divided into control group (N=17, mean age 54±12 years), mild-to-moderate OSAHS group (N=15, mean age 54±12 years) and severe OSAHS group (N=15, mean age 56±13 years). Venous blood was collected, plasma and cells were isolated, the expressions of PD-1 and CTLA-4 on the surface of CD(4)(+)T cells were detected by flow cytometry, and plasma VEGF was measured by enzyme linked immunosorbent assay. Results: The proportion of CD(4)(+)T cells in control group, mild-to-moderate OSAHS group and severe OSAHS group were respectively(38±8)%, (35±8)% and (38±6)% (F=1.228, P>0.05). The expression of CTLA-4 on CD(4)(+)T cells were respectively [1.13 (0.59~1.78)]%, [0.45 (0.16~1.43)]% and [0.87(0.47~1.46)]% (H=2.205, P>0.05). The expression of PD-1 on CD(4)(+)T cells were respectively [4.24 (2.12~6.03)]%, [3.54(2.69~5.09)]% and [3.31(1.67~8.25)]% (H=0.541, P>0.05). The concentrations of VEGF in control group, mild-to-moderate OSAHS group and severe OSAHS group were statistically different [(395.16±87.78) ng/L vs (452.85±107.97) ng/L vs (546.42±199.27) ng/L, F=4.827, P=0.013]. Compared with the control group, VEGF concentration was significantly increased in the severe OSAHS group(P<0.01). VEGF concentration was correlated negatively with the lowest SpO(2) (r (s)=-0.480,P=0.001), but positively with apnea-hypopnea index(r (s)=0.403, P=0.005), oxygen desaturation index (r (s)=0.378, P=0.010) and proportion of SpO(2) less than or equal to 90% of total sleep time(r (s)=0.547, P=0.000 3). Conclusion: There was no significant difference of PD-1 and CTLA-4 expression on CD(4)(+)T cells in patients with and without OSAHS. The expression of VEGF was elevated in OSAHS patients, and increased with the severity of OSAHS and hypoxia.

Stature Estimation of Sichuan Han Females Based on X-ray Photography of Total Lower Limbs.

OBJECTIVES: To establish a mathematical model of stature estimation for Sichuan Han females on the basis of the relationship between lower limbs and individual height, thus to provide evidence for forensic identification. METHODS: Samples were collected from 171 Sichuan Han females. Large flat panel multi-function digital photography system was used to take the full-body X-ray films of the lower limbs. Indexes of long bones and stature of the subjects were measured, respectively. A linear regression analysis was carried out on the correlation between them, and a mathematical model of the stature calculation was established. Then the mathematical model was used to calculate the stature of another 29 Sichuan Han females to test its accuracy. RESULTS: The maximum length of femur （x1） had the highest correlation with stature. A total of 13 linear regression equations were established （P<0.05）, with the correlation coefficient （R） 0.821-0.897 and the standard error of the estimation （SEE） 2.994-3.812 cm. The backtesting showed that the equation y=41.604+1.205 x1+1.318 x6+2.444 x12+1.852 x13-2.388 x14 had the smallest mean absolute deviation （2.485 years old） and the highest accuracy of ±2SEE （92.9%）, and that the equation y=48.783+2.568 x1 had the highest accuracy of ±1SEE （60.7%）. CONCLUSIONS: The stature estimation is high by using the long bones of the lower limbs has high accuracy.

[Stature Estimation for Sichuan Han Female Based on Whole-spine X-ray Photo- graphy].

OBJECTIVES: To establish regression models of stature estimation for Sichuan Han female by the measurement of total vertebral column length in the frontal and lateral X-ray films of whole-spine. METHODS: The frontal and lateral X-ray films of whole-spine were collected from 200 Sichuan Han females by large flat-panel multi-functional universal radiography and fluoroscopy system. The data and mean values of frontal and lateral total vertebral column length were measured and calculated in all the samples, respectively. The relationship of combined multi-markers and stature were analysed by linear regression analysis, and the mathematical models of stature estimation were established. The data of 30 new samples were selected and inputted for verifying the accuracy of the mathematical models. RESULTS: The total vertebral column length showed a good correlation with stature, and the mean values of the frontal and lateral X-ray films of total vertebral column length had the highest correlation coefficients. Three established linear regression equation models were statistically significant （P<0.05）, and the equation established with lateral total vertebral column length showed the highest accuracy. CONCLUSIONS: The stature estimation by the measurement of total vertebral column length has high accuracy.

Evolution of High-Temperature Superconductivity from a Low-T_{c} Phase Tuned by Carrier Concentration in FeSe Thin Flakes.

We report the evolution of superconductivity in an FeSe thin flake with systematically regulated carrier concentrations by the liquid-gating technique. With electron doping tuned by the gate voltage, high-temperature superconductivity with an onset at 48 K can be achieved in an FeSe thin flake with T_{c} less than 10 K. This is the first time such high temperature superconductivity in FeSe is achieved without either an epitaxial interface or external pressure, and it definitely proves that the simple electron-doping process is able to induce high-temperature superconductivity with T_{c}^{onset} as high as 48 K in bulk FeSe. Intriguingly, our data also indicate that the superconductivity is suddenly changed from a low-T_{c} phase to a high-T_{c} phase with a Lifshitz transition at a certain carrier concentration. These results help to build a unified picture to understand the high-temperature superconductivity among all FeSe-derived superconductors and shed light on the further pursuit of a higher T_{c} in these materials.

Induction of apoptosis by capsaicin in hepatocellular cancer cell line SMMC-7721 is mediated through ROS generation and activation of JNK and p38 MAPK pathways.

Capsaicin, one of the major pungent ingredients found in red peppers, has been shown to have anti-carcinogenic effect on various cancer cells through multiple mechanisms. In this study, we investigated the apoptotic effect of capsaicin on human hepatocellular cancer cell line SMMC-7721, as well as the possible mechanisms involved. Treatment of SMMC-7721 cells with capsaicin resulted in a dose-dependent inhibition of cell-viability and induction of apoptosis which was associated with the generation of ROS and persistent disruption of mitochondrial membrane potential. These effects were significantly blocked when cells were pretreated with a general antioxidant N-acetyl cysteine (NAC). We also found that capsaicin induced JNK and p38 MAPK phosphorylation. JNK and p38 MAPK inhibitor effectively blocked capsaicin-induced SMMC-7721 cell apoptosis. In addition, NAC completely blocked phosphorylation of JNK and p38 MAPK induced by capsaicin. Our results indicate that capsaicin induced in SMMC-7721 cell apoptosis through generation of intracellular ROS and activation of JNK and p38 MAPK pathways.

Mitochondrial DNA haplogroups and somatic mutations are associated with lung cancer in patients from Southwest China.

Mitochondrial DNA mutations play crucial roles in the pathogenesis and progression of human malignancies. Therefore, to determine whether maternal background or mitochondrial DNA somatic mutations were essential cofactors in the lung cancer of Chinese patients as well, the complete mitochondrial DNA displacement loop of the primary cancerous, matched para-cancerous normal and distant normal tissues for 79 Chinese patients with lung cancer were analyzed in this study. Our results indicated that the higher detected frequency of haplogroups prevalent in southern East Asia (53.16%; 42/79) versus those of northern East Asia in the studied population supported the southern East Asian characteristics of the Chinese lung cancer group. Further statistical analysis revealed that the haplogroups F* and G* contributed to the susceptibility to lung cancer in Chinese patients. In addition, by comparing sequences from different tissues of the same patients, a total of eight somatic mutations from six patients were detected. Combined with the fourteen somatic mutations identified in our previous study, the somatic mutation spectrum of the 79 Chinese patients with lung cancer was 25.32% (20/79). Our results suggest that mitochondrial DNA haplogroups and somatic mutations are associated with lung cancer in patients from Yunnan, Southwest China, and that somatic mitochondrial DNA mutations in the displacement loop can serve as potential biomarkers for clinical utility.

Deciphering the spectrum of somatic mutations in the entire mitochondrial DNA genome.

The mitochondrion is a crucial intracellular organelle responsible for regulating cellular energy metabolism, producing free radicals, initiating and executing the apoptotic pathways. Previous studies have shown that somatic mutations in mitochondrial DNA are associated with various tumors, which may be involved during carcinogenesis and tumor progression. To examine the mutation pattern in cancer, 625 reported somatic mutations in the mitochondrial DNA genome were analyzed. We found that, except for deletions and insertions, most somatic mutations were point mutations, accounting for 89.44% of somatic mutations. Transition was the predominant form of somatic mutation in the entire mitochondrial DNA genome, accounting for 87.12% of point mutations, most of which were homoplastic. Frequency statistics analysis of point mutations indicated that, except for 3 tRNA genes, the mutations were distributed on all resting genes and in the D-loop region, with the latter showing the highest frequency of somatic mutation (19.34%), followed by the tRNA leucine 2 gene and non-coding regions between base pairs 5892 and 5903, while 13 coding-region genes and 2 rRNA genes showed a relatively lower frequency of somatic point mutations. Nonsynonymous mutations and terminal amino acid changes were the primary point somatic mutations detected from 13 coding-region genes, which may cause mitochondrial dysfunction in cancer cells. We found that the somatic mutations may affect the mitochondrial DNA genome; the non-coding region should be examined to identify somatic mutations as potential diagnostic biomarkers for early detection of cancer.

Local expression and role of BMP-2/4 in injured spinal cord.

We investigated local changes in BMP-2/4 expression in rat spinal cords 1 week following injury to study the damage effects of BMP-2/4 in spinal cord injury (SCI). Sprague Dawley rats (45, 4 months old) were randomized into three groups comprising 15 rats each: a SHAM group, an SCI without noggin group (SCIO), and an SCI with noggin group (SCID). The SCIO and SCID groups were subjected to spinal cord hemisection, and motor activity was assessed using the BBB score. Expression of BMP-2/4 in each injured spinal cord section was examined by hematoxylin and eosin staining, immunohistochemistry, and western blot. There were no significant differences in BBB scores among the three groups (P > 0.05). Following hemisection, the BBB score in the SHAM group was significantly higher than in the other two groups on the 1st day after modeling (P < 0.05), and the BBB scores in the SCIO and SCID groups were not significantly different (P > 0.05). Seven days after modeling, the BBB score in the SHAM group was significantly higher than in the other two groups (P < 0.05), and the BBB score in the SCID group was obviously higher than in the SCIO group (P < 0.05). The expression of BMP-2/4 was highest in the SCIO group and lowest in the SHAM group (P < 0.05). SCI can cause severe impairment of motor activity in rats. Seven days after SCI, the local expression of BMP-2/4 had obviously increased; noggin can effectively inhibit the expression of BMP-2/4 and reduce impairment.

Efficacy of robot-assisted core decompression combined with human umbilical cord-derived mesenchymal stem cell transplantation for osteonecrosis of the femoral head.

OBJECTIVE: The aim of this paper was to evaluate the effects of robot-assisted core decompression combined with human umbilical cord-derived mesenchymal stem cell (hUC-MSC) transplantation for early-stage osteonecrosis of the femoral head (ONFH). PATIENTS AND METHODS: A retrospective analysis was performed on 18 patients with a total of 26 hips who were diagnosed with Association Research Circulation Osseous stage 2 avascular necrosis of the femoral head and who received core decompression combined with hUC-MSC transplantation. All surgeries were completed under robotic assistance. Preoperative and postoperative visual analogue scale (VAS) scores and the Harris Hip Score (HHS) were recorded to assess clinical function. A Magnetic Resonance Imaging (MRI) examination was performed at the last follow-up. RESULTS: The mean follow-up was 18.6 months (12-28 months), the VAS score (4.5±0.8 vs. 0.9±0.2, t=12.6, p≤0.001) and HHS (79.5±5.8 vs. 60.5±4.6, t=14.3, p≤0.001) were significantly improved at the last follow-up, compared with preoperative value. The MRI results showed that the necrotic volume of the femoral heads was significantly reduced. CONCLUSIONS: Robot-assisted core decompression combined with hUC-MSC transplantation is a feasible and relatively safe method for the treatment of femoral head necrosis.

Targeting of GSK-3ß by miR-214 to facilitate gastric cancer cell proliferation and decrease of cell apoptosis.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Targeting of GSK-3ß by miR-214 to facilitate gastric cancer cell proliferation and decrease of cell apoptosis, by H.-L. Li, S. Liang, J.-H. Cui, G.-Y. Han, published in Eur Rev Med Pharmacol Sci 2018; 22 (1): 127-134-DOI: 10.26355/eurrev_201801_14109-PMID: 29364479" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/14109.

Increased miR-142 and decreased DJ-1 enhance the sensitivity of pancreatic cancer cell to adriamycin.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Increased miR-142 and decreased DJ-1 enhance the sensitivity of pancreatic cancer cell to adriamycin, by G.-Y. Han, J.-H. Cui, S. Liang, H.-L. Li, published in Eur Rev Med Pharmacol Sci 2018; 22 (22): 7696-7703-DOI: 10.26355/eurrev_201811_16390-PMID: 30536312" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/16390.

Long noncoding RNA DLX6-AS1 functions as a competing endogenous RNA for miR-577 to promote malignant development of colorectal cancer.

Since this article has been suspected of research misconduct and the corresponding authors did not respond to our request to prove originality of data and figures, "Long noncoding RNA DLX6-AS1 functions as a competing endogenous RNA for miR-577 to promote malignant development of colorectal cancer, by F.-R. Zhou, Z.-P. Pan, F. Shen, L.-Q. Huang, J.-H. Cui, K. Cai, X.-L. Guo, published in Eur Rev Med Pharmacol Sci 2019; 23 (9): 3742-3748-DOI: 10.26355/eurrev_201905_17800-PMID: 31115000" has been withdrawn. The Publisher apologizes for any inconvenience this may cause. https://www.europeanreview.org/article/17800.

Long noncoding RNA DLX6-AS1 functions as a competing endogenous RNA for miR-577 to promote malignant development of colorectal cancer.

OBJECTIVE: Recent researches have proved that long noncoding RNAs (lncRNAs) play essential roles in tumorigenesis. The aim of this study was to investigate the exact role of lncRNA DLX6-AS1 in the development of colorectal cancer (CRC), and to explore the possible mechanism. PATIENTS AND METHODS: DLX6-AS1 expression in CRC tissues was detected by Real Time-quantitative Polymerase Chain Reaction (RT-qPCR). Function assays were conducted to detect the effect of DLX6-AS1 on the proliferation and metastasis of CRC in vitro. Furthermore, luciferase reporter gene assay and RNA immunoprecipitation assay (RIP) were used to explore the underlying mechanism of DLX6-AS1. RESULTS: DLX6-AS1 expression in CRC samples was significantly higher than that of adjacent tissues. Loss of DLX6-AS1 markedly inhibited the proliferation, migration, and invasion of CRC cells. Furthermore, luciferase reporter gene assay and RIP assay showed that DLX6-AS1 acted as a competing endogenous RNA via sponging miR-577 in CRC. CONCLUSIONS: DLX6-AS1 could promote the proliferation, migration, and invasion of CRC by sponging miR-577, which might offer a potential therapeutic target for CRC.

Increased miR-142 and decreased DJ-1 enhance the sensitivity of pancreatic cancer cell to adriamycin.

OBJECTIVE: Phosphatidylinositol-3 kinase (PI3K)/protein kinase B (AKT) signaling pathway is related to tumorigenesis by up-regulating survivin. Phosphatase and tensin homologue deleted on chromosome ten (PTEN) can suppress PI3K/AKT signaling pathway, while DJ-1 is the negative regulator of PTEN. DJ-1 up-regulation is closely correlated with the occurrence, progression, and drug resistance of pancreatic cancer. MicroRNA-142 (MiR-142) is significantly declined in pancreatic cancer tissue. Bioinformatics analysis demonstrated that complementary binding site exists between miR-142 and DJ-1. This investigation, therefore, aimed to study the role of miR-142 in the regulation of DJ-1-PTEN/PI3K/AKT/Survivin signaling pathway as well as in pancreatic cancer cell proliferation, apoptosis, and adriamycin (ADM) resistance. MATERIALS AND METHODS: Dual luciferase assay was performed to assess the targeted relationship between miR-142 and DJ-1. MiR-142, DJ-1, and PTEN expressions in SW1990 cells and drug-resistant SW1990/ADM cells were compared. SW1990/ADM cells were divided into five groups, including mimic-NC, miR-142 mimic, small interfere normal control (si-NC), si-DJ-1, and miR-142 mimic + si-DJ-1 groups. DJ-1, PTEN, phosphorylated-AKT (p-AKT), and Survivin expressions were tested. Cell apoptosis was determined by flow cytometry. Cell proliferation was evaluated by EdU staining. RESULTS: MiR-142 targeted inhibited DJ-1 expression. MiR-142, PTEN, and cell apoptosis significantly down-regulated, while DJ-1, p-AKT, Survivin, and cell proliferation significantly elevated in SW1990/ADM cells compared with SW1990 cells. MiR-142 mimics and/or si-DJ-1 transfection markedly reduced DJ-1, p-AKT, and Survivin expressions enhanced PTEN level, attenuated cell proliferation, enhanced cell apoptosis, and weakened ADM resistance. CONCLUSIONS: MiR-142 over-expression weakened ADM resistance in pancreatic cancer cells by targeting DJ-1 to enhance PTEN expression and attenuate PI3K/AKT signaling pathway activity.

Targeting of GSK-3ß by miR-214 to facilitate gastric cancer cell proliferation and decrease of cell apoptosis.

OBJECTIVE: Wnt/ß-catenin pathway regulates cell proliferation and apoptosis. GSK-3ß degrades ß-catenin and negatively regulates Wnt/ß-catenin pathway. A previous study indicated that the GSK-3ß expression was significantly reduced in gastric cancer, along with the increase of miR-214 expression. Bioinformatics analysis revealed complementary binding sites between miR-214 and 3'-UTR of GSK-3ß mRNA. This study investigated the regulatory role and related mechanism of miR-214 in the proliferation and apoptosis of gastric cancer cells. PATIENTS AND METHODS: Gastric cancer tissues were collected from patients and the expressions of miR-214, GSK-3ß and ß-catenin were determined. Dual luciferase reporter gene assay was used to study the regulatory role between miR-214 and GSK-3ß. Expressions of miR-214, GSK-3ß, ß-catenin and survivin from GES-1 and MKN-28 cells were detected. Flow cytometry was used to measure cell proliferation and apoptosis. In vitro cultured MKN-28 cells were treated with miR-214 inhibitor and/or pSicoR-GSK-3ß. Levels of GSK-3ß, ß-catenin and survivin were detected, cell apoptosis was evaluated by flow cytometry and proliferation was tested by EdU staining. RESULTS: Compared to normal gastric mucosa, the levels of miR-214 and ß-catenin were elevated, and the expression of GSK-3ß was decreased in gastric cancer tissues. Compared to GES-1 cells, the expressions of miR-214, ß-catenin and survivin in MKN-28 cells were upregulated, along with downregulation of GSK-3ß expression. The proliferation was enhanced whilst apoptosis was suppressed. After the transfection of miR-214 inhibitor and/or pSicoR-GSK-3ß, GSK-3ß expression was induced in MKN-28 cells while ß-catenin and survivin expressions were inhibited, along with the increase of cell apoptosis. CONCLUSIONS: MiR-214 decreases GSK-3ß expression and promotes the pathogenesis of gastric cancer. The inhibition of miR-214 reduces the proliferation of gastric cancer cells via upregulation of GSK-3ß and suppression of Wnt/ß-catenin signal pathway, which provides fundamental support for the future therapy of gastric cancer.

CD44V6 regulates gastric carcinoma occurrence and development through up-regulating VEGF expression.

OBJECTIVE: Gastric carcinoma (GC) is one of the most common malignant tumors around the world. It is featured as high morbidity, poor prognosis, and short survival, thus seriously threats to the quality of life. The mechanism of GC is still unclear, leading to a difficulty in the treatment. CD44V6 plays an important role in tumorigenesis and progression, while its role in GC still needs further elucidation. PATIENTS AND METHODS: GC tissue and para-carcinoma tissue were collected from patients in different tumor-mode-metastasis (TNM) stages. CD44V6 and vascular endothelial growth factor (VEGF) expressions were detected by real-time PCR and Western blot. Their correlations with the clinicopathological characteristics of GC were analyzed. GC cell line SGC-7901 was cultured in vitro and divided into control, scramble group, and CD44V6 small interfering RNA (siRNA) group. Cell proliferation was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay. Cell apoptosis was evaluated by caspase 3 activity assay. RESULTS: CD44V6 and VEGF protein expressions significantly increased in GC tissue compared with adjacent normal control (p < 0.05). CD44V6 expression was correlated with differentiation, lymph node metastasis, and TNM staging (p < 0.05). CD44V6 was positively correlated with VEGF (p < 0.05). CD44V6 siRNA reduced CD44V6 and VEGF expressions in SGC-7901, inhibited cell proliferation, and enhanced caspase 3 activity compared with control (p < 0.05). CONCLUSIONS: CD44V6 participates in GC occurrence and development by up-regulating VEGF expression. Targeting CD44V6 regulates GC progression through inhibiting VEGF expression, promoting cell apoptosis, and restraining cell proliferation.

[Study on antibiotic resistance of Escherichia coli and Enterococcus colonized in intestine of neonates from neonatal intensive care unit].

Objective: To understand the antibiotic resistance of bacteria colonized in intestine of the neonates from neonatal intensive care unit (NICU) and provide evidence to guide clinical antibiotic treatment. Methods: From May, 2014 to May, 2015, a total of 572 stool samples were collected from the neonates of NICU in our hospital. Escherichia coli and Enterococcus were detected with VITEK-2 system. Results: A total of 328 strains of E. coli and 243 strains of Enterococcus were isolated respectively in this study. The 199 strains of E. coli selected for drug susceptibility test showed lower resistant rate to imipenem, ertapenem, amikacin, nitrofurantoin, ranging from 0.50% to 3.52% and showed higher resistant rate to ampicillin, tetracycline, trimethoprim/sulfamethoxazole and cefazolin, ranging from 54.27% to 84.92%. No meropenem resistant strainsere were found. The percentage of ESBLs production strains was 45%. The multi drug resistance test showed that 34.6% of the strains were resistant to four antibiotics. Three strains were resistant to seven antibiotics. The 243 strains of Enterococcus showed lower resistant rate to quinupristin/dalfopristin, nitrofurantoin, streptomycin, ranging from 0.41% to 4.53% and showed higher resistant rate to ampicillin, benzylpenicillin, ciprofloxacin, tetracycline, gentamicin and erythromycin, ranging from 70.78% to 91.77%. No strains which were resistant to tigecycline, vancomycin, rina thiazole amine/ketone were found. The multi drug-resistance test showed that 86.5% of the strains were resistant to five antibiotics. Conclusions: According to the analysis of the 199 strains of E. coli and 243 strains of Enterococcus isolated from the neonates, we found that the resistance of intestinal bacteria in the neonates was very serious, showing multi drug resistance. It is necessary to use antibiotics according to the drug susceptibility test results in clinical treatment.

[Study on identification of Cronobacter spp. species in 11 areas in China].

OBJECTIVE: To compare different methods on the identification of Cronobacter (C.) spp. species and to choose an optimum one. METHODS: Biochemical test, 16S rRNA and fusA sequencing methods were carried out. RESULTS: When using the biochemical test, 105 strains showed six different conditions but C. turicensis and C. universalis could not be effectively identified. Under the 16S rRNA sequencing analysis, all the strains were divided into 5 groups but C. sakazakii and C. malonaticus were tangled. Finally, all the strains were identified into 58 C. sakazakii, 30 C. malonaticus, 11 C. dublinensis, 5 C. turicensis, 1 C. muytjensii, under the fusA sequencing analysis. CONCLUSION: Currently, fusA sequencing analysis seemed an effective method for identifying the species of Cronobacter. Since fusA sequencing analysis method was less intuitive, another method for rapid testing should be developed.

Orthotopic transplantation model of human gastrointestinal cancer and detection of micrometastases.

AIM: To establish a relevant animal model of human gastrointestinal cancer, which can be used for repetitive investigations, so as to improve our understanding and management of carcinogenesis and cancer metastasis. METHODS: Intact tissues of human colorectal and pancreatic cancers were transplanted in nude mice. The biological characteristics of the original and the corresponding transplanted tumors were investigated by HE staining, PAS staining and immunostaining. The metastases in the livers and lungs of nude mice were investigated by immunostaining with biotinylated mab KL-1 and by RT-PCR using CK20 specific primers. RESULTS: There were totally 9 of 16 surgical specimens growing in nude mice subcutaneously and/or orthotopically (4 of 6 colorectal and 5 of 10 pancreatic cancer). Tumor cell content of the specimens and freezing of tissue specimens are important factors influencing the growth of transplanted tumor. In the group of fresh tumor tissues with greater than 50% tumor cell content, the success rate of the transplantation was 100% (3 cases of pancreatic cancer and 3 cases of colorectal cancer). The orthotopically trans-planted tumors resemble the original tumor morphologically and biologically, including TAA expression such as CEA by immunohistochemistry, and CEA level in the serum of mice. Ki-67 labeling index and the expression of TAA especially K-ras, 17-1A and RA-96, are associated with the potential of tumor growth in nude mice. Micrometastases in the lungs and livers of tumor bearing mice can be detected by immunostaining with biotinylated mab KL-1 and CK20-specific RT-PCR. CONCLUSION: An orthotopic transplantation model for human colon and pancreatic cancer in nude mice has been set up. We have also established sensitive detection methods with CK-immunohistochemistry and CK20-RT-PCR to study xenotransplanted human cancer and its metastatic cancer cells in the liver and lung of nude mice. This study may be helpful in understanding the mechanism of cancer metastasis and in developing new diagnostic methods and therapeutic strategies for metastases including micrometastases.

Controlled release of dual drug-loaded hydroxypropyl methylcellulose matrix tablet using drug-containing polymeric coatings.

A dual drug-loaded hydroxypropylmethylcellulose (HPMC) matrix tablet simultaneously containing drug in inner tablet core and outer coated layer was formulated using drug-containing aqueous-based polymeric Eudragit RS30D dispersions. Effects of coating levels, drug loadings in outer layers, amount and type of five plasticizers and talc concentration on the release characteristics were evaluated on the characteristics in simulated gastric fluid for 2 h followed by a study in intestinal fluids. Melatonin (MT) was selected as a model drug. The surface morphology of dual drug-loaded HPMC tablets using scanning electron microscope (SEM) was smooth, showing the distinct coated layer with about 75-microm coating thickness at the 15% coating level. Unlike the uncoated and conventionally coated HPMC tablet, the dual drug-loaded HPMC matrix tablet gave a biphasic linear release, showing a zero-order for 4 h (first) followed by another zero-order release when fitted using linear regression (r(2) = 0.99). As the coating levels (15, 25%) increased, the release rate was further decreased. The biphasic release profiles of dual drug-loaded HPMC matrix tablet was unchanged except when 25% coating level containing 0.5% drug concentration was applied. As the drug concentration in polymeric coating dispersion increased (0.25-1.0%), the amount of drug released increased. The time for the first linear release was also advanced. However, the biphasic release pattern was not changed. The biphasic release profiles of dual drug-loaded HPMC matrix tablet were highly modified, depending on the amount and type of five plasticizers. Talc (10-30%) in coating dispersion as an anti-sticking material did not affect the release profiles. The current dual drug-loaded HPMC matrix tablet, showing biphasic release profiles may provide an alternative to deliver drugs with circadian rhythmic behaviors in the body but needs to be further validated in future in human studies. The dual drug-loaded coating method is also interesting for the modified release of poorly water-soluble drugs because solubilizers and other additives can be added in drug-containing polymeric coating dispersions.