[Exploration of the clinical application of combined endoscopic and laparoscopic surgery in early gastric cancer: 15 cases].

Objective: To investigate the application of combined gastroscopy and laparoscopy (dual scope) in the treatment of early gastric cancer. Methods: In this descriptive case series study, we retrospectively collected data on 15 patients with cT1b stage gastric cancer who had undergone combined laparoscopic and endoscopic surgery in the 900th Hospital of the People's Liberation Army of China from May 2020 to October 2022. The study cohort comprised nine men and six women of median age 59 (range: 47-76) years and median body mass index 20.9 (range: 18.3-26.2) kg/m2. Seven of the lesions were located on the lesser curvature of the gastric antrum and eight in the gastric angle. All lesions were biopsied for pathological examination and evaluated by endoscopic ultrasonography, followed by endoscopic submucosal dissection (ESD) and laparoscopic regional lymph node dissection. Studied variables included surgical and pathological features, postoperative factors, and outcomes. Results: In this group of patients, the median (range) operative time for ESD was 45 (30-82) minutes, the duration of laparoscopic lymph node dissection (45.1±8.6) minutes, and the median (range) intraoperative blood loss during lymph node dissection 30 (10-80) mL. Of the 13 patients with negative postoperative horizontal margins, four were stage SM1 and had no lymph node metastases (Stage SM1) and nine were Stage SM2, of which had one positive regional lymph node and two received additional standard distal gastrectomy with D2 lymphadenectomy concurrently because of positive ESD specimens (lymph node negative). No lymph node metastases were found in the surgical specimens of these patients. The remaining two patients had positive vertical margins; both had undergone concurrent standard distal gastrectomy with D2 lymphadenectomy. One of them was found to be lymph node positive (No. 3, one node). Four patients had impaired gastric emptying after dual-scope treatment, all of whom recovered well with symptomatic management; one patient with a suspected lymphatic leak was also managed conservatively. There were no cases of postoperative bleeding, abdominal infection, or incisional infection. At a median follow-up of 14 (6-26) months, no tumor recurrence or metastasis had been identified in any of the patients. Three patients had a grade B nutrition score 3 to 6 months after surgery, all of whom had undergone major gastrectomy, and two patients who had undergone dual-scope surgery reported an increase in acid reflux and belching after surgery compared with the preoperative period. Conclusion: A combined technique is safe and feasible for the treatment of early gastric cancer and is worthy of further exploration.

[Factors affecting long-term survival of advanced high-grade serous ovarian cancer].

Objective: To identify the factors associated with long-term survival and guide the decision for primary surgery in patients with advanced high-grade serous ovarian cancer(HGSOC). Methods: In this case-control study, clinical parameters, including surgical and non-surgical associated factors, were collected and compared between the patients with short-term (<2 years) and long-term (>5 years) survival who all underwent primary debulking surgery (PDS) followed by carboplatin and paclitaxel chemotherapy from January 2004 to December 2016. Univariate analysis was examined by chi-square test and multivariate analysis was performed by logistic regression analysis. Results: There were 95 cases long-term survival (LTS group) and 77 cases short-term survival (STS group) in 698 newly diagnosed HGSOC patients with International Federation of Gynecology and Obstetrics (FIGO) stage Ⅲc and Ⅳ who met include and exclude criteria. (1) Univariate analysis showed that the proportion of complete cytoreduction with no visible residual disease (R0) at PDS and platinum sensitivity in LTS group were significantly higher than those in STS group (P<0.01). The surgical complexity score (SCS), the preoperative serum CA125 level and the ascites volume in the LTS group were significantly lower than those of the STS group (all P<0.05). In the LTS group, the preoperative incidence of lesions in retrograde peritoneum of the bladder, serosal and mesangial membrane of the small intestine, upper abdominal peritoneum and liver parenchyma were significantly lower than those in the STS group (all P<0.05). Multivariate logistic regression analysis showed that platinum sensitivity (OR=0.016, 95%CI: 0.004-0.063, P<0.01), ascites volume >500 ml (OR=3.193, 95%CI: 1.285-7.930, P=0.012), and SCS ≥8 (OR=17.433, 95%CI: 2.281-133.25, P=0.003) were independent factors affecting long-term survival (P>0.05). (2) Totally 37 of 95 in long-term survival and 16 of 77 in short-term survival achieved R0 cytoreduction at PDS. Univariate analysis showed that preoperative serum CA125 level, preoperative lesion score, preoperative lesion (DS) score, ascites volume, platinum sensitivity,and SCS were significantly correlated with the R0 PDS (all P<0.05). Multivariate analysis showed that ascites volume >500 ml (OR=5.199, 95%CI: 2.015-13.409, P=0.001), DS >2 (OR=15.264, 95%CI: 5.843-39.874, P<0.01) and SCS ≥4 (OR=4.176, 95%CI: 1.618-10.777, P=0.003) were independent factors associated with R0 cytoreduction. In patients with DS ≤2 or SCS <4, but not those with DS >2 or SCS ≥4, R0 cytoreduction was significantly associated with long-term survival. Conclusion: The intrinsic biology of tumor is the factor influencing long-term survival of advanced HGSOC patients, and those who present with wide intraperitoneal metastases and need to remove multiple organs may not benefit from R0 cytoreduction.

MIR22HG inhibits cell growth, migration and invasion through regulating the miR-24-3p/p27kip1 axis in thyroid papillary carcinomas.

OBJECTIVE: To explore the underlying mechanism of ncRNA (MIR22HG) in thyroid papillary carcinomas. PATIENTS AND METHODS: 40 pairs of thyroid papillary carcinomas tissues and adjacent normal tissues were collected from patients of the First Affiliated Hospital of Guangxi Medical University, who underwent oral surgery. qRT-PCR was applied to detect the expression of MIR22HG, miR-24-3p and p27kip1 in tissues and cells. Western blot was used to measure the protein level of p27kip1 in tissues and cells. Kaplan-Meier plot was used to analyze the overall survival rates in thyroid papillary carcinomas. Pearson's correlation analysis was used to analyze the correlation relationship among MIR22HG, miR-24-3p and p27kip1 expression. Flow cytometric assay was applied to measure cell apoptosis. Transwell assay was used to assess cell migration and invasion abilities. Luciferase reporter assay was applied to verify the molecular relationships among MIR22HG, miR-24-3p and p27kip1 in thyroid papillary carcinomas. RESULTS: LncRNA MIR22HG and p27kip expressed low while miR-24-3p expressed high in thyroid papillary carcinomas and cells. Overexpression of MIR22HG inhibited cell proliferation, migration and invasion, whereas promoted cell apoptosis in thyroid papillary carcinomas cells. However, these effects were reversed by upregulation of miR-24-3p. Further exploration showed that the promoted effects of miR-24-3p mimics on thyroid papillary carcinomas cells were suppressed by enhancing p27kip1 expression. Meanwhile, MIR22HG induced p27kip1 expression by binding miR-24-3p in thyroid papillary carcinomas. CONCLUSIONS: MIR22HG inhibited cell growth through modulating p27kip1 by decreasing miR-24-3p expression in thyroid papillary carcinomas, providing a new modulate mechanism and therapeutic targets in thyroid papillary carcinomas.

Genomic and transcriptional heterogeneity of multifocal hepatocellular carcinoma.

BACKGROUND: Hepatocellular carcinoma (HCC) often presents with multiple nodules within the liver, with limited effective interventions. The high genetic heterogeneity of HCC might be the major cause of treatment failure. We aimed to characterize genomic heterogeneity, infer clonal evolution, investigate RNA expression pattern and explore tumour immune microenvironment profile of multifocal HCC. PATIENTS AND METHODS: Whole-exome sequencing and RNA sequencing were carried out in 34 tumours and 6 adjacent normal liver tissue samples from 6 multifocal HCC patients. Protein expression of Ki67, AFP, P53, Survivin and CD8 was detected by immunohistochemistry. Fluorescence in situ hybridization was carried out to validate the amplification status of sorafenib-targeted genes. RESULTS: We deciphered genomic and transcriptional heterogeneity among tumours in each multifocal HCC patient including mutational profiles, copy number alterations, tumour evolutionary trajectory and tumour immune microenvironment profiles. Of note, sorafenib-targeted alterations were identified in the trunk of phylogenetic tree in only one out of the six patients, which may explain the relative low treatment response rate to sorafenib in clinical practice. Moreover, we demonstrated RNA expression patterns and tumour immune microenvironment profiles of all nodules. We found that RNA expression pattern was associated with Edmondson-Steiner grading. Based on the differential expression of 66 reported immune markers, unsupervised hierarchical clustering analysis of 34 nodules identified immune subsets: one low expression cluster with seven nodules and one high expression cluster with 11 nodules. CD8+ T cells were more enriched in nodules of the high expression cluster. CONCLUSIONS: Our study provided a detailed view of genomic and transcriptional heterogeneity, clonal evolution and immune infiltration of multifocal HCC. The heterogeneity of druggable targets and immune landscape might help interpret the clinical responsiveness to targeted drugs and immunotherapy for multifocal HCC patients.

[Screening of microRNAs targeting Notch signaling pathway implicated in inner ear development and the role of microRNA-384-5p].

Objective: To screen the key microRNAs targeting Notch signaling pathway in inner ear and investigate its potential regulating function. Methods: The interaction network and the Core-Notch network, involved with key genes in Notch signal pathway and differential-expressed microRNAs in inner ear, were constructed by bioinformatics methods. The important microRNAs in regulating Notch signaling pathway were screened via topological and GO analysis, followed by in vivo and in vitro investigation. Results: MiRNA-384-5p was identified as a key regulator specifically expressed in mouse brain and inner ear, which could down-regulate Notch1. The Notch1 expression was found significantly down-regulated in miRNA-384-5p-mimic-transfected HeLa cells. The dual-luciferase reporter gene assay further confirmed the effect of miRNA-384-5p on the down-regulation of Notch1 and Dll4 in Notch signaling pathway. Conclusions: The Core-Notch network is constructed to screen microRNAs implicated in inner ear development, and miRNA-384-5p is screened and verified to be target-regulating the Notch signaling pathway, which could be the potential target in the regeneration of impaired hair cells.

[A study of the diagnostic value of endobronchial ultrasound guide sheath transbronchial lung biopsy combined with virtual bronchoscopic navigation in peripheral pulmonary lesions].

OBJECTIVE: To evaluate the diagnostic value of endobronchial ultrasound guide sheath transbronchial lung biopsy (EBUS-GS-TBLB) combined with virtual bronchoscopic navigation (VBN) in peripheral pulmonary lesions (PPLs). METHODS: Cases with a PPL identified by computed tomography in Affiliated Hospital of Medical College of Ningbo University underwent EBUS-GS-TBLB with or without VBN randomly between Nov. 2014 to Aug. 2015. X-ray guidance was not performed in these cases. The sensitivity and the operation time were evaluated in the 2 groups. RESULTS: A total of 184 patients were enrolled and completed this study. Among them 117 were males and 67 were females. There were 93 cases in the group of EBUS-GS-TBLB with VBN, and 91 in the group without VBN. The diagnostic sensitivity of VBN group was 72.04%(67/93). Among these positive cases, 64.1% cases (43/67) were malignant tumors, and 35.9% cases (24/67) were benign lesions. The mean operation time was (45±10)min. In the group without VBN, the diagnostic sensitivity was 69.23%(63/91), including 33 malignant tumors(52.4%, 33/63), and 30 benign lesions(47.6%, 30/63). The mean operation time was (55±10)min. There was no significant difference between EBUS-GS-TBLB with VBN group and EBUS-GS-TBLB without VBN group in diagnostic sensitivity (χ(2)=0.175, P=0.747). But there was a significant difference in the mean operation time between the 2 groups (t=6.522, P<0.001). EBUS-GS-TBLB was well tolerated. No severe procedure-related complications such as pneumothorax and hemoptysis were observed. CONCLUSION: VBN cannot improve the diagnostic sensitivity, but it can clear the location of lesion, and shorten the operation time. This technique helps to abandon the X-ray guidance. EBUS-GS-TBLB combined with VBN is a safe and effective technique for PPLs.

[Value of the albumin to globulin ratio in predicting severity and prognosis in myasthenia gravis patients].

OBJECTIVE: To assess the predictive value of the albumin to globulin ratio (AGR) in evaluation of disease severity and prognosis in myasthenia gravis patients. METHODS: A total of 135 myasthenia gravis (MG) patients were enrolled between February 2009 and March 2015. The AGR was detected on the first day of hospitalization and ranked from lowest to highest, and the patients were divided into three equal tertiles according to the AGR values, which were T1 (AGR <1.34), T2 (1.34≤AGR≤1.53) and T3 (AGR>1.53). The Kaplan-Meier curve was used to evaluate the prognostic value of AGR. Cox model analysis was used to evaluate the relevant factors. Multivariate Logistic regression analysis was used to find the predictors of myasthenia crisis during hospitalization. RESULTS: The median length of hospital stay for each tertile was: for the T1 21 days (15-35.5), T2 18 days (14-27.5), and T3 16 days (12-22.5) (P<0.01), and Kaplan-Meier curves showed significant difference among the three groups. In the univariate model, serum albumin, creatinine, AGR and MGFA clinical classification were related to prognosis of myasthenia gravis. At the multivariate Cox regression analysis, the AGR (P<0.001) and MGFA clinical classification (P<0.001) were independent predictive factors of disease severity and prognosis in myasthenia gravis patients. Respectively, the hazard ratio (HR) were 4.655 (95% CI: 2.355-9.202) and 0.596 (95% CI: 0.492-0.723). Multivariate Logistic regression analysis showed the AGR (P<0.001) and MGFA clinical classification were related to myasthenia crisis. CONCLUSION: The AGR may represent a simple, potentially useful predictive biomarker for evaluating the disease severity and prognosis of patients with myasthenia gravis.

Medial plantar flap to repair defects of palm volar skin.

PURPOSE: The methods of repairing defects in fingers, volar skin of the palm and soft tissue were investigated. METHODS: From 2010 to the present, we examined 12 cases in which medial plantar skin flaps were used to repair defects in the fingers and palm. According to skin and soft tissue defects in the fingers and palm, a flap was designed using the medial plantar artery as the vessel pedicle. The flap was dissected and isolated between the abductor hallucis and flexor digitorum brevis, and transplanted to the hand. We then observed the skin colour, skin texture and tactile sensitivity of the hand, as well as the shape and function of the foot. RESULTS: Follow-up for 3-28 months showed that the flaps survived in all 12 cases, with soft skin, healthy appearance, colour consistent with the palm skin and no pigmentation. The two-point discrimination was 5-7 mm. The donor foot functioned well, and the scar at the donor site was slight and had an aesthetic appearance. CONCLUSION: The free medial plantar flap is an ideal flap for repairing soft tissue defects in the palm.

Pharmacokinetic of four probe drugs in adriamycin-induced nephropathy rat.

BACKGROUND AND AIM: Probe drugs have been widely used to assess the activities of various CYP450 (cytochromes P450) isoenzymes in many fields of drug metabolism and pharmacogenetics. The nephrotic syndrome characterized by massive proteinuria and hypoproteinemia, whether that would influence the pharmacokinetics of probe drugs or not is still unclear. The purpose of the study was to investigate the pharmacokinetic of four probe drugs in adriamycin (ADR)-induced nephropathy rat. MATERIALS AND METHODS: The rats were randomly divided into Control-group (n = 10) and ADR-group (n = 10). Nephrotic syndrome was established by weekly injections of ADR for 2 weeks. After dynamic monitoring of 24-h total urinary protein for 4 weeks, we confirmed that nephrotic syndrome had developed. The rats were administered intragastrically with phenacetin, tolbutamide, omeprazole and bupropion (15, 5, 15, and 15 mg/kg, respectively). The blood samples were determined by LC-MS (Liquid Chromatography-Mass Spectrometry) method. RESULTS: The pharmacokinetics parameter of tolbutamide in ADR-group and Control-group were AUC(0-t) 15.371 ± 4.107, 6.901 ± 5.738 (mg/L*h), MRT(0-t) 8.751 ± 0.754, 6.032 ± 0.63 (h), t1/2 3.88 ± 0.423, 3.602 ± 0.693 (h), Tmax 6.2 ± 3.768, 1.95 ± 0.798 (h), CL/F 0.038 ± 0.005, 0.107 ± 0.037 (L/h/kg), V/F 0.212 ± 0.043, 0.567 ± 0.258 (L/kg), Cmax 1.853 ± 0.384, 1.422 ± 1.312 (mg/L). There was statistical difference in AUC, MRT, CL, V and Tmax of tolbutamide between two groups (p < 0.05), but no pharmacokinetics difference for phenacetin, bupropion and omeprazole. CONCLUSIONS: The pharmacokinetics of tolbutamide was changed in ADR-induced nephropathy rat. It is not suitable for tolbutamide to evaluate the activity of CYP450 in nephrotic syndrome.

The application of ultrasonic harmonic scalpel in the radical surgery of gastric cancer.

PURPOSE: Ultrasonic harmonic scalpel has been widely applied to laparoscopic surgery of gastric cancer, but it has not been evaluated properly in open surgery. The objective of this study was to evaluate the value of the ultrasonic harmonic scalpel in the open radical surgery of gastric cancer. METHODS: 106 gastric cancer patients who had accepted distal D2 lymphadenectomy were included in this study. Patients were divided into ultrasonic harmonic scalpel (UHS) surgery group (50 cases) and conventional electric scalpel surgery group (56 cases). UHS surgery group patients were accepted surgery by ultrasonic harmonic scalpel. Instead, conventional electric scalpel surgery group patients were accepted surgery by monopolar electrocautery shovel and other traditional instruments. Then the average operation time, intra-operative blood loss, number of harvested lymph nodes, average post operative drainage within 3 days, and postoperative hospital stay were collected and compared between the two groups. RESULTS: The average operation time, blood loss, postoperative hospital stay in UHS group were significantly lower than traditional group (P < 0.05). The number of lymph node dissection was significantly higher than conventional surgery group (P < 0.05). There were no difference between two groups in average drainage within 3 days after surgery and the hospitalization costs (P > 0.05). In the presence of atherosclerotic patients, the average operation time, blood loss in UHS surgery group were significantly lower than the traditional group (P < 0.05). CONCLUSION: Ultrasonic harmonic scalpel may have better effect in the radical surgery of gastric cancer patients. It meets the requirements of the future development of precise surgical procedure.

Association analysis between the -2518MCP-1(A/G) polymorphism and generalized aggressive periodontitis in a Chinese population.

BACKGROUND AND OBJECTIVE: It has been suggested that aggressive periodontitis has a genetic basis. Monocyte chemoattractant protein 1 (MCP-1) plays a critical role in the recruitment of monocytes and the development of periodontitis. The -2518MCP-1(A/G) polymorphism has been implicated as a risk or susceptibility factor for a variety of autoimmune conditions and inflammatory diseases. The intent of this investigation was to study whether the -2518MCP-1(A/G) polymorphism is associated with generalized aggressive periodontitis in the Chinese population. MATERIAL AND METHODS: One hundred and twenty-four patients with generalized aggressive periodontitis and 94 healthy subjects were included in this case-control study. Genomic DNA was isolated from a peripheral blood sample obtained from each subject. Gene polymorphisms of -2518MCP-1(A/G) were analyzed by a standard polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. A logistic regression analysis was performed to test the association between the -2518MCP-1(A/G) genotype (alleles) and generalized aggressive periodontitis with adjustment of the major covariates (gender, age and smoking status). RESULTS: There was no significant association of the -2518MCP-1(A/G) polymorphism with generalized aggressive periodontitis in the unstratified subjects. However, when patients were stratified by gender, the frequency of the G(+) genotype was significantly lower in female patients with generalized aggressive periodontitis compared with female controls (p = 0.036, adjusted odds ratio = 0.3, 95% CI: 0.1-0.9). In female patients with generalized aggressive periodontitis, the probing pocket depth was larger in subjects with the AA genotype than in subjects with the G(+) genotype (5.07 mm vs. 4.30 mm; Z = -2.470, p = 0.014). CONCLUSION: The polymorphisms of -2518MCP-1 may play an important role in determining generalized aggressive periodontitis susceptibility in this cohort of Chinese women.

Family-based association analysis of S100A8 genetic polymorphisms with aggressive periodontitis.

BACKGROUND AND OBJECTIVE: It is known that S100A8, a member of the S100 calcium-binding protein family, is associated with inflammatory diseases, including periodontitis. Our previous population-based study found an association between two polymorphisms, rs3795391 (A > G) and rs3806232 (A > G), in the upstream region of the S100A8 gene and aggressive periodontitis (AgP) in Chinese people. Based on those results, this investigation set out to analyze and corroborate whether the association also exists within families. MATERIAL AND METHODS: Two hundred and four subjects from 73 nuclear families were recruited. All probands and their relatives were diagnosed according to the 1999 classification of periodontal diseases. Anticoagulated peripheral blood samples were collected from all the subjects, and DNA was extracted. The two single nucleotide polymorphisms (SNPs; rs3795391 and rs3806232) were detected and analyzed by standard polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay. Analysis of genotype/allele was performed by Family-Based Association Test (FBAT) software ( http://www.biostat.harvard.edu/~fbat/default.html). RESULTS: There was a statistically significant association of the SNP rs3795391 with AgP in the additive genetic model (chi(2) = 3.9836, d.f. = 1, p = 0.0459). Allele A showed significantly preferential transmission to the AgP affected individuals (Z = 1.996, p = 0.0459). The other SNP, rs3806232, showed no significant results in all models. CONCLUSIONS: This family-based association study supports the previous findings that SNP rs3795391 (A > G) of the S100A8 gene might contribute to AgP susceptibility. This is, to our knowledge, the first investigation about AgP using FBAT in genetic analysis.

Failure behavior of coated nickel-based superalloy under thermomechanical fatigue.

Thermomechanical fatigue (TMF) is a major life limiting factor for gas turbine blades. In this study, the failure behavior of NiCrAlY overlay coated nickel-based superalloy M963 was investigated under out of phase (OP) TMF. All tests were carried out under mechanical strain control with a cyclic period of 200 s. Results revealed that the fatigue life of high velocity oxygen fuel spraying (HVOF) coated specimen was longer than that of air plasma spraying (APS) coated one, but shorter than that of bare superalloy M963 at a given strain range. It was found that cracking process in the APS coating was different from that in the HVOF coating, which was shown by the sketches to understand the crack initiation and propagation behavior.

Elevation of C-reactive protein and interleukin-6 in plasma of patients with aggressive periodontitis.

BACKGROUND AND OBJECTIVE: Systemic levels of C-reactive protein and interleukin-6 have been reported to be elevated in patients with periodontitis compared with periodontally healthy individuals. Most studies included patients with chronic periodontitis and comprised predominantly Caucasians. The aim of this study was to determine the relative levels of C-reactive protein and interleukin-6 in plasma of patients with aggressive periodontitis in China and to examine the relationships between these two inflammatory mediators and clinical parameters, peripheral blood cells and protein variables. MATERIAL AND METHODS: Plasma samples were collected from 84 patients with aggressive periodontitis and from 65 control subjects. Periodontal examination consisted of taking probing depth and attachment loss measurements. The levels of plasma C-reactive protein and interleukin-6 were determined using enzyme-linked immunosorbent assays. RESULTS: The levels of plasma C-reactive protein in patients with aggressive periodontitis were significantly higher than those in control subjects (1.87 vs. 0.52 mg/L). The level of plasma interleukin-6 in patients with aggressive periodontitis was 1.20 pg/mL, higher than that in control subjects (0.08 pg/mL). Multiple linear regression analysis showed that log C-reactive protein was significantly related to severe sites percentage and albumin following correction for age, gender, body mass index and smoking (p = 0.000, p = 0.008, respectively). Log interleukin-6 was found to be significantly correlated with periodontal diagnosis, leukocyte count and level of fasting blood glucose after adjusting for the confounders (p = 0.000, p = 0.009 and p = 0.013, respectively). CONCLUSION: Patients with aggressive periodontitis have significantly elevated levels of plasma C-reactive protein and interleukin-6. These elevated inflammatory factors might potentially increase the risk for cardiovascular events and glucose dysregulation in relatively young individuals.

WITHDRAWN: Adjuvant portal vein chemotherapy improves the efficacy of hepatectomy for hepatocellular carcinoma with portal vein tumor thrombus.

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Telomerase antisense inhibition for the proliferation of endometrial cancer in vitro and in vivo.

The objective of this study was to investigate the antitumor effect of antisense telomerase oligodeoxynucleotides to endometrial cancer cells in vitro and in vivo. Antisense oligodeoxynucleotides (ODNs) against the human telomerase transcripatse (hTERT) synthesized to serve as telomerase inhibitors. Reverse transcription-polymerase chain reaction and 3-[4,5-dimethylthiazol-2yl]-2,5-diphenyltetrazolium bromide (MTT) assay were used to test the expression of hTERT messengerRNA (mRNA) and inhibition of cell proliferation in vitro. In vivo, antitumor effects of ODNs or combined with cisplatin were evaluated in endometrial cancer xenograft. Telomerase activity was tested by telomeric repeat amplification protocol. Antisense ODNs could inhibit proliferation of human endometrial cancer cells (HEC-1-A) in vitro, and downregulate the expression hTRET mRNA in a dose- and period-dependent manner. The tumor growth inhibitory rate of low- and high-dose ODNs were 34.20% and 89.21%, and combined group was 75.30%. Telomerase activity was downregulated to 87.32% compared to the control in the ODNs-treated xenograft tumors. Antisense oligonucleotides of hTERT effectively inhibit the growth of endometrial cancer cell line. Telomerase inhibitor might be a new strategy for chemotherapy or chemoprevention in endometrial cancer.

Induction of cytotoxic T-lymphocyte responses using dendritic cells transfected with hepatocellular carcinoma mRNA.

This study aims to induce an efficient expansion of cytotoxic T-lymphocytes (CTL) from peripheral blood mononuclear cells (PBMCs) using dendritic cells (DC) transfected with hepatocellular carcinoma (HCC) messenger RNA (mRNA) for adoptive immunotherapy of HCC. Dendritic cells are generated from PBMCs. HCC mRNA is isolated either from HepG-2 cells or from tumour tissue from three HCC patients, and then amplified using the polymerase chain reaction (PCR). Expansion of CTLs is achieved from PBMCs induced by DCs transfected with HCC mRNA and cytotoxicity is measured using a crystal violet staining assay. The proportion of CD3+, CD4+ and CD8+ cells is determined using flow cytometry. Dendritic cells transfected with the total HCC mRNA stimulated antigen-specific cytotoxic T-cell responses that are capable of recognising and killing autologous tumour cells in vitro. The cytotoxic activity was inhibited by treatment with anti-CD3, anti-CD8 and anti-MHC class I monoclonal antibodies, but not with anti-CD4 and MHC class II antibodies. In conclusion, HCC mRNA-transfected DCs may represent a broadly applicable vaccine strategy to induce potentially therapeutic CTL responses in HCC.

[Protective effect of tumor necrosis factor-alpha on motor neurons after peripheral nerve injury].

OBJECTIVE: To investigate the protective effect of tumor necrosis factor-alpha(TNF-alpha) on spinal motor neurons after peripheral nerve injury. METHODS: Twenty Wistar rats were divided into two groups, the right sciatic nerves of 20 Wistar rats were transected, the proximal stumps were inserted into a single blind silicone tube. 16 microliters of normal saline(NS) and TNF-alpha(30 U/ml) were injected into the silicone tubes. After 2 weeks, the 4th, 5th lumbar spinal cord were taken for examination. Enzyme histochemical technique and image analysis were used to show acetylcholinesterase(AChE) and nitric oxide synthase(NOS) activity of spinal motor neurons. RESULTS: The number of AChE and NOS staining neurons were 8.65 +/- 1.98 and 5.92 +/- 1.36 in the experimental group and 6.37 +/- 1.42 and 8.67 +/- 1.45 in the control group respectively, there were significant difference between the two groups(P < 0.01). CONCLUSION: It suggests that TNF-alpha has protective effect on motor neurons after peripheral nerve injury.

Development of nonionic surfactant/phospholipid o/w emulsion as a paclitaxel delivery system.

Paclitaxel is an anticancer agent with low aqueous solubility. More extensive clinical use of this drug is somewhat delayed due to lack of appropriate delivery vehicles. An attempt was made to adopt an o/w emulsion as the drug carrier which incorporated paclitaxel in the triacylglycerol stabilized by a mixed-emulsifier system. A suitable formulation was found in this study: 0.75 mg/ml paclitaxel, 10% (w/v) oil blend, 4% (w/v) EPC, 3% (w/v) Tween 80 in 2.25% (w/v) glycerol solution. The formulated emulsion has very good stability when stored at 4 degrees C, and the paclitaxel containment efficiency can be maintained above 95% and the mean emulsion diameter around 150 nm for at least 3 months. Paclitaxel-emulsion displayed cytotoxicity against HeLa cells with IC50 at 30 nM. The average life span of ascitic-tumor-bearing mice was prolonged significantly by the treatment of paclitaxel-emulsion (P<0.05). The formulated emulsion is a promising carrier for paclitaxel and other lipophilic drugs.