Percutaneous Image-Guided Microwave Ablation for Treating Postsurgical Intrapulmonary Oligometastases or Oligorecurrence.

OBJECTIVE: To assess the clinical efficacy of microwave ablation (MWA) in treating tumour patients with postsurgical intrapulmonary oligometastases or oligorecurrence (PIORO). STUDY DESIGN: Descriptive study. Place and Duration of the Study: Departments of Thoracic Surgery and Oncology, Jinan Central Hospital and Qilu Hospital, Jinan, China, from January 2014 to June 2023. METHODOLOGY: Clinical data of 31 patients with PIORO receiving treatment with MWA were retrospectively analysed. After undergoing MWA, the patients were followed up for computed tomography (CT) examination on the 7th day, 1st month, and every 3 months, respectively. The Kaplan-Meier method was conducted to evaluate the clinical outcomes; overall survival (OS), progression-free survival (PFS), and time to local progression (TTLP). RESULTS: All patients with PIORO were successfully treated with MWA. The 3-year survival rate of patients was 35.5%. The median OS was 26.0 months, the median PFS was 11.1 months, and the median TTLP was 14.4 months. Patients with oligometastatic or oligorecurrent tumours ≤3 cm in diameter showed better PFS (≤3 cm, 14.261 m vs. >3 cm, 7.786 m; p <0.01) and TTLP (≤3 cm, 19.522 m vs. >3 cm, 12.214 m; p <0.05) than those with tumours >3 cm in diameter. Clinical characteristics of the patients were not significantly correlated with OS. CONCLUSION: MWA, as a topically therapeutic method, is an effective procedure for tumour patients with PIORO, especially in cases of oligometastatic or oligorecurrent tumours ≤3 cm in diameter. KEY WORDS: Microwave ablation, Thermal ablation, Oligometastases, Oligorecurrence, Progression-free survival, Survival.

Effect of dental plaque control on infection of Helicobacter pylori in gastric mucosa.

BACKGROUND: Data on the role of dental plaque in the transmission of Helicobactor pylori have varied. Furthermore, there has been few reports on the relationship between dental plaque control and H. pylori infection of gastric mucosa. The purpose of this study was to elucidate this potential relationship. METHODS: The 13C urea breath test was conducted on 56 subjects who received dental plaque control and 51 subjects who did not. RESULTS: The prevalence of H. pylori in the gastric mucosa was 19.64% in patients who received dental plaque control, which was significantly lower than in those without dental plaque control (84.31%). CONCLUSION: Long-term professional dental plaque control was associated with less gastric reinfection by H. pylori, suggesting that dental plaque control may help to prevent H. pylori-induced gastric disease or reinfection.

Automatic phantom-less QCT system with high precision of BMD measurement for osteoporosis screening: Technique optimisation and clinical validation.

BACKGROUND: Currently dual-energy X-ray absorptiometry (DXA) and phantom-based quantitative computed tomography (PB-QCT) have been utilized to diagnose osteoporosis widely in clinical practice. While traditional phantom-less QCT (PL-QCT) is limited by the precision of manual calibration using body tissues, such as fat and muscle. OBJECTIVE: The aim of this study is to validate the accuracy and precision of one newly-developed automatic PL-QCT system to measure spinal bone mineral density (BMD) and diagnose osteoporosis. METHODS: A total of 36 patients were enrolled for comparison of BMD measurement between DXA and QCT. CT images of 63 patients were analyzed by both PB-QCT and newly developed automatic PL-QCT system, then the BMD results generated by the automatic PL-QCT were utilized to diagnose osteoporosis. The diagnostic outcomes were compared with that of DXA and PB-QCT to assess the performance of the new system. RESULTS: BMD test results showed that the automatic PL-QCT system had higher precision than previous studies performed with QCT, while maintaining similar capability to diagnose osteoporosis as DXA and PB-QCT. Area under curve (AUC) result of PL-QCT was larger than 0.8 for predicting spine DXA T-score in receiver operating characteristic (ROC) analysis. Pearson correlation analysis (r â€‹= â€‹0.99) showed strong linear correlation and Bland-Altman analysis (bias â€‹= â€‹3.0mg/cc) indicated little difference between the two methods. The precision result (CV â€‹= â€‹0.89%) represented good reproducibility of the new system. CONCLUSION: The traditional PL-QCT system has relatively low reproducibility due to the manual selection of the region of interest (ROI) of body tissues. Automatic selection of ROI in this new system makes the BMD testing more convenient and improves precision significantly. Compared with traditional BMD measurement methods, the automatic PL-QCT system had higher precision in accurate diagnosis of osteoporosis with great potential in translational research and wide clinical application. TRANSLATIONAL POTENTIAL STATEMENT: With high accuracy and precision, the automatic PL-QCT system could serve as an opportunistic screening tool for osteoporosis patients in the future. It could also facilitate related researches by providing more reliable data collection, both retrospectively and longitudinally.

The susceptibility of three strains of Chinese minipigs to diet-induced type 2 diabetes mellitus.

Researchers have extensively used animal models to study diabetes mellitus. In this study, the authors determined the susceptibility of three strains of Chinese minipigs to diet-induced type 2 diabetes mellitus. For 8 months, the researchers fed Nongda control minipigs (n=4) a normal diet and fed Bama, Wuzhishan and Nongda minipigs (n=6 per group) a high-sucrose, high-fat diet. They measured the minipigs' body weights, fasting serum glucose concentrations and insulin concentrations each month. Every 2 months, they measured serum triglyceride, cholesterol, high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) levels and carried out intravenous glucose tolerance tests (IVGTTs). The Bama and Wuzhishan minipigs were relatively susceptible to diabetes induced by the high-sucrose, high-fat diet, though susceptibility differed among individual animals in the same strain. On the other hand, Nongda minipigs were relatively resistant to diet-induced diabetes. These results provide a foundation for diabetes-related genetic analyses in minipigs with high and low susceptibility to diet-induced type 2 diabetes.

Coconut-based activated carbon fibers for efficient adsorption of various organic dyes.

In this study, using coconut fibers as raw material, activated carbon fibers were prepared via carbonization and KOH activation processes. The morphology, composition, specific surface area, pore structure and thermal stability of the resulting activated carbon fibers were systematically characterized. It was found that the activation process increases the specific surface area of carbon fibers to a greater extent via formation of a large number of micropores (0.7-1.8 nm) and a certain amount of slit-shaped mesopores (2-9 nm). The specific surface area and the pore volume of the activated carbon fibers reach 1556 m2 g-1 and 0.72 cm3 g-1, respectively. The activation process can also decompose the tar deposits formed after the carbonization process by pyrolysis, making the surface of the activated carbon fibers smoother. To study the adsorption properties of the as-prepared activated carbon fibers, the adsorption capacities and adsorption kinetics of various organic dyes including methylene blue, Congo red and neutral red were investigated. The adsorption capacities of the dyes increased with the increasing initial dye concentrations, and varied greatly with the pH value of the system. In methylene blue and neutral red systems, the adsorption capacities reach the maximum at pH 9, and in the Congo red system, it reaches the maximum at pH 3. The adsorption capacities of the activated carbon fibers in methylene blue, Congo red and neutral red systems reached equilibrium at 150, 120, and 120 min, and the maximum adsorption capacities were 21.3, 22.1, and 20.7 mg g-1, respectively. The kinetics of the adsorption process was investigated using three models including pseudo-first-order, pseudo-second-order and intraparticle diffusion models. The results indicated that the dynamic adsorption processes of coconut-based activated carbon fibers to methylene blue, Congo red and neutral red were all in accordance with the second-order kinetic model, and the equations are as follows: t/Q t = 0.1028 + t/21.3220, t/Q t = 0.1128 + t/21.5982 and t/Q t = 0.0210 + t/20.6612.

Trans-arterial gene therapy for hepatocellular carcinoma in a rabbit model.

AIM: To study the effect of adenovirus (Ad)-p53 gene therapy on hepatocellular carcinoma (HCC) in a rabbit model. METHODS: VX2 tumor was grown in the liver of 24 rabbits. Animals were divided into four groups: group A receiving trans-arterial gene therapy (Ad-p53) only, group B receiving combined Ad-p53 therapy and trans-arterial embolization (lipiodol), group C receiving trans-arterial chemoembolization (lipiodol + mitomycin C), control group (D) receiving sodium chloride. Tumor volume (V1) was measured by using MRI (d 13). Interventional procedure was applied (d 14). Tumor volume (V2) was assessed by MRI (d 21) and the mean ratio (V2/V1) was calculated. After the second MRI, specimens of the liver were abstained and examined immunohistochemically using mutant-type p53 antibody. The positive expression was scored. RESULTS: Compared with control group (chi= 3.14 +/- 0.64), therapeutic groups all showed a significant decrease in the tumor growth ratio (P<0.05). A slight difference was found between group A (chi = 2.35 +/- 0.59) and group B (chi = 1.75 +/- 0.28) (P=0.048). No statistically significant difference was observed between group B and group C (chi = 2.00 +/- 0.44). The positive expression rate of mutant-type p53 was the lowest in group B and significantly different between group A and group C (P<0.05). Compared to the control subjects, groups A and C both showed a decrease in the expression of mutant-type p53, but there was no significant difference between them. CONCLUSION: Trans-arterial Ad-p53 gene therapy can reduce tumor growth of HCC in rabbit model.

[The application of METRx microendoscopes diskectomy system in the treatment of far lateral lumbar disc herniation].

OBJECTIVE: To introduce the METRx microendoscopes diskectomy system in the treatment of far lateral lumbar disc herniation. METHODS: Fourteen cases of far lateral lumbar disc herniation were operated with METRx from February 1999 to December 2002. Among them, the average age was 49 years old (range 41 - 55 years old), male in 10 cases, female in 4 cases. All cases were single disc herniation; L(4), 5 herniation in 6 discs, L(5)-S(1) herniation in 8 discs; foraminal disc herniation in 6 cases, extra-foraminal disc herniation in 8 cases. RESULTS: All the cases were followed up from 12 to 46 months (average 26.5 months) with the results of excellence in 10 cases, good in 3 cases, fair in 1 case and no failure case. There were no disc infection, dura laceration, nerve root injury and herniation recurrence. CONCLUSIONS: METRx is suitable for far lateral lumbar disc herniation with the advantages of minimal invasive, complete decompression of nerve root and rapid recovery. The correct approach and precise surgical technique are the key points for this operation.

[The role of artificial disc replacement in restoring lumbar segment motion and intervertebral height].

OBJECTIVE: To investigate the role of artificial disc replacement (ADR) in restoring lumbar segment motion and intervertebral height (IH). METHODS: Seventy-five patients with intervertebral disc diseases underwent artificial disc replacement and were divided into 3 groups to be followed up for 2 years (group A, n = 32), 3 years (group B, n = 27), and 5 years (group C, n = 19) respectively. Before the operation and during the follow-up the range of motion (ROM) of the involved lumbar segments and IH were assessed. RESULTS: The mean ROM and IH of the group A were 15 degrees and 14.1 mm respectively during the follow-up 2 years after operation. The mean ROM and IH of the group B were 14.2 degrees and 13.8 mm respectively during the follow-up 2 years after the operation and 13.7 degrees and 13.4 mm respectively during the follow-up 3 years after operation. The mean ROM and IH of the group C were 9.1 degrees and 11.5 mm respectively during the follow-up 2 years after the operation and 8.8 degrees and 11.0 mm respectively during the follow-up 3 years after operation, and 8.0 degrees and 10.3 mm respectively during the follow-up 5 years after operation. No significant differences in ROM and IH were found among different groups at any time point (all P > 0.05). Malposition was found in immediately after operation in 11 patients, all of the group C. During the follow-up, 3 cases of prostheses subsidence and 3 cases of annular ossification were found. No severe complications appeared. CONCLUSION: ADR is effective in restoring the ROM and IH of vertebrae suffering from intervertebral disc diseases.

[Design and experiment of the pedicle aiming device for the cervical spine].

OBJECTIVE: To probe the accuracy and safety of using an aiming device in the transpedicular fixation of cervical spine. METHODS: Eight cervical specimens were obtained. We used the computed tomography (CT) to scan C3 to C7, measured the medial angle of the pedicle, and determined the location of the pedicle projecting on the articular process. Then we took the oblique X-ray film, measured the cephalic/caudal angle of the pedicle, and determined the location of the pedicle projecting on the articular process. All the specimens were equally divided into two groups. Screws of 2.8 mm x 30 mm, were used. Specimens in one group were inserted with the transpedicular screw manually, while specimens in the other one inserted with the transpedicular screw using a self-designed aiming device that can be modulate at the three dimensions according to the angles of the pedicles. RESULTS: The first group totally had 40 screws from C3 to C7. There were 13 screws in the pedicle, 9 violated the walls of the pedicle but not involved the adjunct structure, and 18 injured the important structure such as spinal cord, verteberal artery, or nerve root. In the other group, only 4 screws violated the walls of the pedicle but not involved the adjunct structure, and the others all in the pedicles. The difference was of statistical significance (P < 0.01). CONCLUSION: In the cervical spine, transpedicular fixation using an aiming device can improve the accuracy and safety during operation.

[Pharmacodynamic tests of wulong kangai on transplantable tumors in mice].

OBJECTIVE: To observe the pharmacodynamic and side effects of Wulong Kangai, a new drug of Chinese traditional herbal medicine, on 4 strains of mice transplantable tumors. METHOD: Mice transplantable tumors S180, H22, P388 and Lewis were used in the pharmacodynamic test on the granules of Wulong Kangai. The test on each tumor strain was repeated three times. In each test, 50 mice were used and divided into 5 groups. They were negative control group treated by physiological saline, cyclophosphamide control group and 3 test groups treated respectively with Wulong Kangai at deferent dosages of 10, 25, 40 g x kg(-1) x d(-1) in the treatment of Lewis and P388 and 15, 30, 50 g x kg(-1) x d(-1) in the treatment of S180 and H22. RESULT: The tumor weight were inhibited at the rates of 90.1%, 30.8%, 49.8% and 52. 3% in the mice with tumors of Lewis, P388, S180, and H22 by high dosage of Wulong Kangai as compared with negative control group. The inhibitory rates in cyclophosphamide groups were 90.6%, 77.2%, 79.6% and 60.3% respectively. The mice body weights grew slower in high dose groups treated by Wulong Kangai granule. CONCLUSION: Wulong Kangai was effective in treating mice transplantable tumors of Lewis, P388, S180 and H22 with a dose-dependent manner. The Lewis was the most sensitive strain to the drug among the 4 kinds of tested tumors. Side effects appeared during 9-11 days of uninterrupted treatment with high dose Wulong Kangai.

[Study on the mitochondrion DNA mutation in tumor tissues of gynecologic oncology patients].

OBJECTIVE: To investigate the mitochondrion DNA (mtDNA) mutation in tumor tissues of gynecologic oncology patients and their relationship to tumorigenesis and tumor development. METHODS: The samples of tumor tissue and their proximal normal tissue of 32 gynecological malignant tumor patients as well as 8 gynecological benign tumor patients were taken. The malignants were 5 squamous cervical carcinomas, 10 endometrial carcinomas and 17 epithelial ovarian cancers (EOC). The benign tumors were 4 ovarian epithelial tumors and 4 uterine myomas. Polymerase chain reaction-single strain conformation polymorphism (PCR-SSCP) and DNA sequencing were done to examine mtDNA mutation. RESULTS: The mtDNA mutation rate and polymorphism rate were 68.8% and 56.3% respectively in 32 cases of malignants. The mtDNA mutation rate and polymorphism rate were 2/8 and 4/8 respectively in 8 cases of benigns. The difference between malignants and benigns was significant (P < 0.05). The hot point of mtDNA mutation were located in Cytb gene region. The characteristic mtDNA mutation is multigene and multisites mutation. The mojarity (63.6%) of patients involved in 2 more gene mutation. CONCLUSIONS: There is high frequent mtDNA coding area mutation in gynecological malignant tumors, indicating that mtDNA coding region mutation is closely related to gynecological malignants development and progression. It may be an important extra nuclear molecular genetic alteration in mechanisms of tumorigenesis.

The clinical value of combined use of MR imaging and multi-slice spiral CT in limb salvage surgery for orthopaedic oncology patients: initial experience in nine patients.

BACKGROUND: The purpose of this prospective study was to evaluate the value of the combined use of MR imaging and multi-slice spiral CT for limb salvage surgery in orthopaedic oncology patients. PATIENTS AND METHODS: Nine consecutive patients with lower/upper limb malignant bone tumours (7 osteosarcomas and 2 chondrosarcomas) were treated with limb-salvaging procedures. Preoperative planning including determination of the osteotomy plane and diameters of the prosthesis was performed basing on the preoperative CT and MR images. The histopathology was performed as golden diagnostic criteria to evaluate the accuracy of CT and MR-based determination for tumour's boundary. RESULTS: The tumour extension measured on MRI was consistent with the actual extension (P>0.05, paired Student's t test), while the extension measured on CT imaging was less than the actual extension. The length, offset and alignment of the affected limb were reconstructed accurately after the operation. An excellent functional outcome was achieved in all patients. CONCLUSIONS: In the present study, MRI was found to be superior to CT for determining the tumour extension, combined use of MRI and CT measurement provided high precision for the fit of the prosthesis and excellent functional results.

OPG knockout mouse teeth display reduced alveolar bone mass and hypermineralization in enamel and dentin.

Recent studies showed that local injection or upregulation of OPG gene would result in early temporal retardation of tooth development. It was assumed that this retardation might cause defective tooth mineralization and pulp formation as the long-term effects. However, since those OPG treatments were transient, any possible long-term effects of OPG addition could not be assessed previously. In the present study, a high-resolution microCT was used to evaluate the long-term effect of OPG gene deprivation on the mineralization and morphology of mouse tooth. Our results showed that the mineralization of alveolar bone in OPG(-/-) mouse tooth was decreased while those of enamel and dentin were increased, compared with the wild-type (WT) group. The labial and lingual dentin thicknesses of OPG(-/-) group were significantly higher and with larger area in enamel and dentin than those of WT group. The size of pulp chamber was also substantially decreased in OPG(-/-) mouse incisor. Different responses in mineralization and morphogenesis to OPG gene deprivation were found between bone and tooth. These effects may be independent of the early odontogenesis, and further studies are warranted to investigate the molecular mechanism of the effect of OPG gene expression on bone formation and later tooth development.

Effect of dental plaque control on infection of Helicobacter pylori in gastric mucosa.

BACKGROUND: Data on the role of dental plaque in the transmission of Helicobacter pylori have varied. Furthermore, there has been few reports on the relationship between dental plaque control and H. pylori infection of gastric mucosa. The purpose of this study was to elucidate this potential relationship. METHODS: The (13)C urea breath test was conducted on 56 subjects who received dental plaque control and 51 subjects who did not. RESULTS: The prevalence of H. pylori in the gastric mucosa was 19.64% in patients who received dental plaque control, which was significantly lower than in those without dental plaque control (84.31%). CONCLUSION: Long-term professional dental plaque control was associated with less gastric reinfection by H. pylori, suggesting that dental plaque control may help to prevent H. pylori-induced gastric disease or reinfection.

[Clinical observation on injection of small dose of morphine into Neiguan (PC 6) for treatment of chest pain in the patient with acute myocardial infarction].

OBJECTIVE: To probe into an effective injection way of Morphine for treatment of chest pain of acute cardiac infarction. METHODS: Ninety cases of myocardial infarction were randomly divided into 3 groups, an acupoint-injection group, an intravenous injection group and a hypodermic injection group, 30 cases in each group. The acupoint-injection group were treated with injection of 2 mg Morphine into bilateral Neiguan (PC 6) respectively, and the intravenous injection group with intravenous injection of 5 mg Morphine and the hypodermic injection group with hypodermic injection of 5 mg Morphine, and other treatments were same in the 3 groups. The analgesic effects were assessed with visual analogue scale (VAS) 5, 10, 30, 60 and 180 minutes after treatment and the complications were observed. RESULTS: There were no significant differences among the 3 groups before treatment in the VAS score (P > 0.05). The analgesic effect in the acupoint-injection group was better than those in other two groups 5 min, 30 min and 180 min after treatment (all P < 0.01). The incidence rate of nausea and vomiting of 0.3% in the acupoint-injection group was significantly lower than 40.0% in the intravenous injection group and 20.0% in the hy podermic injection group (all P < 0.01). CONCLUSION: Injection of small dose of Morphine into Neiguan (PC 6) has a definite therapeutic effect on chest pain of acute myocardial infarction with earlier analgesic effect, smaller dose of Morphine, longer analgesic duration and less complications.

[Association of 86 bp variable number tandem repeat polymorphism of interleukin-1 receptor antagonist gene with lumbar disc disease].

OBJECTIVE: To investigate the association of 86 bp variable number tandem repeat (VNTR) polymorphism of interleukin-1 receptor antagonist (IL-1Ra) gene with lumbar disc disease and intervertebral disc degeneration. METHODS: The 86 bp VNTR polymorphism of IL-1Ra gene was analyzed with PCR and electrophoresis for 81 patients with lumbar disc disease and 101 volunteers without sciatica (control). The grade of intervertebral disc degeneration was determined with magnetic resonance imaging, and the association of 86 bp VNTR polymorphisms with lumbar disc disease and intervertebral disc degeneration in those younger than 45 years was assessed. RESULTS: The presence of 86bp VNTR polymorphisms of IL-1Ra gene was detected in both patients with lumbar disc disease and the control subjects. The distribution of 86 bp VNTR polymorphisms of IL-1Ra gene showed no significant difference between the two groups, but the distributions of 1/1, 1/2 and 2/2 or 1, 2 genotypes differed significantly. The current data did not support a significant association between the distribution of IL-1Ra gene 86bp VNTR polymorphism and lumbar disc degeneration. CONCLUSIONS: IL-1Ra gene 86bp VNTR polymorphism is present among Chinese population in association with lumbar disc disease, but not with lumbar disc degeneration.

[The post-injury regenerating ability of temporomandibular joint of rats exposed to the repeated + Gz forces].

OBJECTIVE: To observe the post-injury regenerating ability of temporomandibular joint of rats exposed to the repeated + Gz forces. METHODS: One hundred and eight male rats were randomly assigned to 9 groups, with 12 rats in each group. Group A was normally fed serving as the blank control group. Group B was only fixed with rat-kept devices for 5 minutes as the fixed control group. Group C was borne +1 Gz for 5 minutes. In the same position as group C, group D, E, F, G, H and I were repeatedly exposed to +10 Gz for 30 seconds each time with 5 times a day and +1 Gz for 1 minutes as a interval. The 6 groups were treated in the condition above for 4 days a week and 4 weeks in total. The rats in group A, B, C and D were killed on the next day after the centrifuge. The rats in group E, F, G, H and I were killed at 1, 2, 4, 8 and 12 weeks after the centrifuge, respectively. After the rats were killed, temporomandibular joint were taken for observation under optical microscope, scanning electron-microscope (SEM) and transmission electron-microscope (TEM). RESULTS: There was no significant difference observed in group A, B and C. But in group D, E, F, G, H and I pathologic injuries and self-regeneration were seen in TMJ in different degrees. CONCLUSIONS: The injury of TMJ could regenerate gradually with time.

[Screening and identification of ovarian carcinomas related genes].

BACKGROUND & OBJECTIVE: Ovarian cancer is the leading cause of death among gynecological malignancies. Up to now, little is known about specific tumor-suppressor genes or oncogenes involved in the ovarian cancer genesis. Thus isolation of new candidate genes and characterization of their role in ovarian cancer genesis will be helpful for understanding the molecular mechanisms and developing protocols for early diagnosis and therapy of ovarian carcinomas. This study was developed to screen and identify genes related to ovarian carcinomas. METHODS: Modified mRNA differential display PCR and reverse Northern dot blot analysis were used to screen and identify different displayed genes between ovarian carcinoma tissue and normal ovarian tissue. The genes were sequenced and analyzed by bioinformatics software. With these gene fragments as probes, in situ hybridization was used to characterize specific gene expression difference between ovarian cancer tissues and normal ovarian tissues. RESULTS: Of 12 differentially expressed genes, 5 were novel genes, 3 were chromosome genomic repeat sequences and the other 4 were known human genes. Furthermore, in situ hybridization analysis indicated that four genes, ZNF361, PSMA2, OCRC13 (a novel gene on chromosome 1) and OCRC4 (a novel gene on chromosome 9) were highly expressed in 36 samples of ovarian cancer tissues but not in 16 samples of normal ovarian tissues (P< 0.05). More interestingly, the gene OCRC4, with 99% sequence homology to mouse spindlin, a member of gene family specially expressed during gametogenesis, was highly expressed in 19 samples of ovarian cancer from total 36 cases (53%), but not expressed in control samples. CONCLUSION: Our observations indicate that the four genes ZNF361, PSMA2, OCRC4, and OCRC13 may be ovarian cancer related genes. It is speculated that novel gene OCRC4 might also be a member of specific gene family, which plays roles during early period of embryo development and tumorigenesis.

Aberrant expression and deletion of FHIT gene in leukemias.

FHIT (fragile histidine triad) gene at chromosome 3p14.2 usually expresses at a very low level in human tissue and cells. A high frequency of abnormalities in FHIT gene has been demonstrated in various cancers. FHIT is proposed as a putative tumor-suppressor gene. To evaluate the expression of the FHIT gene in various leukemias, bone marrow or peripheral blood samples from 98 leukemia patients were tested by RT-PCR: 38 from patients with AML-[M(2)(9), M(3)(12), M(4)(8), M(5)(9)], 16 with ALL, and 34 with CML-[CP(20), AP(4), BC(10)] of various FAB types, as well as 10 patients with other hematological malignancies. To detect a deletion in sequencing the FHIT gene, the representative aberrant PCR products were cloned and then sequenced. The results showed that 22/38 (58%) patients with AML, 9/16 (56%) patients with ALL and 19/34 (56%) patients with CML were detectable of aberrant FHIT mRNA transcripts or deletion of FHIT. In 6 (16%) AML patients, 3 (19%) ALL patients, and 5 (15%) CML patients, the wild-type product was absent. Some patient's samples - 6 (42%) AML, 6 (38%) ALL, and 14 (41%) CML revealed aberrant FHIT transcripts in addition to a normal-sized band. Samples from healthy donors (PB, n = 12; BM, n = 5) did not indicate any abnormal expression. Eleven isolated fragments from various patterns of FHIT gene expression were investigated using cDNA sequencing. Sequence analysis revealed deletion of exon 4-8, exon 5-8, and exon 5-6 in various leukemias, as well as the deletion of the full FHIT gene sequence. The fused transcripts included: exon 3 and exon 9, exon 3 and exon 7, exon 4 and exon 9, exon 5 and exon 7. Sequence analysis of aberrant fragments present in samples from an AML and a CML patients was detected for point mutations and insert mutations located in exons 2, 8 and 10, plus a variety of aberrant transcripts. Deletion or aberrant FHIT mRNA transcripts in 50/98 (51%) leukemia patients were found. All samples with aberrant FHIT lacked gene product. A Kaplan-Meier plot of survival in patients with AML in relation to FHIT expression revealed that aberrance or loss of FHIT gene significantly correlated with a low clinical remission rate and poor overall survival.

[Soluble expression of peptide containing MUC1/Y-specific epitope in Escherichia coli and preparation of the antibody].

MUC1 mucin is a high molecular weight, type I transmembrane glycoprotein. High and aberrant expression of MUC1 is observed in various types of tumors, which make it an ideal target for tumor biotherapy as well as a biomarker for tumor diagnosis and prognosis. MUC1/Y is an isoform of MUC1 generated by alternative splicing. Specific expression of MUC1/Y in breast cancer as well as its involvement in tumor cell signal transduction have been reported. In order to purify peptides containing MUC1/Y-specific epitope in E. coli and prepare MUC1/Y-specific antibody, DNA fragment encoding the MUC1/Y-specific peptide was amplified by PCR using MUC1/Y full length cDNA as the template and cloned into fusion expression vector pGEX-2T, resulting pGEX-Y30. DNA sequencing was performed to confirm the correct amplification and orientation of the target sequence. Competent E. coli DH5alpha was transformed with pGEX-Y30 and the expression was induced for 4-5 hours in 0.2 mmol/L IPTG at 30 degrees C and 37 degrees C. Expressed proteins were released from the cells by ultrasonication or B-PER II reagent treatments. The fusion protein GST-Y30 were purified by affinity and anion exchange columns and identified by SDS-PAGE and Western-blotting. Polyclonal antibody was prepared by immunizing rabbits with the GST-Y30 protein for 4 times with intervals of 3 weeks and purified by GST column. Western blotting, ELISA and immunohistochemistry analysis were carried out using the purified antibody to confirm its MUC1/Y-binding capacity and specificity. The expressed fusion protein GST-Y30 is about 31 kD in size and represented about 20% of total cellular proteins. The majority of the GST-Y30 protein existed as soluble form when the induction was carried out at both 30 degrees C and 37 degrees C. After the two-step purification, the purity of GST-Y30 was about 94%. The titer of polyserum generated by GST-Y30 immunization was 1:320,000 by ELISA. The antiserum showed MUC1/Y specificity and can recognize MUC1/Y on MCF7 cell. The MUC1/Y-specific polyclonal antibody can be used for studying the role of MUC1/Y in carcinogenesis.