Neuroprotection by intravenous transplantation of bone marrow mononuclear cells from 5-fluorouracil pre-treated rats in a model of ischemic stroke.

Our previous studies showed that bone marrow mononuclear cells (BMMNCs) from 5-fluorouracil (5-FU) pre-treated rats (named BMRMNCs) had a better therapeutic efficacy in ischemia/reperfusion rats as compared to BMMNCs from untreated rats. This study was undertaken to further explore the potential mechanisms underlying the neuroprotective effects of BMRMNCs in the same model. Rats were intravenously pre-treated with 5-FU, and BMRMNCs were collected 7 days later and subjected to flow cytometry for detection of CD34, CD45 and CD90. Middle cerebral artery occlusion (MCAO) was induced in rats, and BMMNCs and BMRMNCs were independently transplanted via the tail vein at 24 h after MCAO. NISSL staining was performed 14 days after cell transplantation and the viable cells in the hippocampus were counted. Stromal cell-derived factor 1 (SDF-1) mRNA expression was detected in the penumbra at 7 and 14 days after treatment. The contents of pro-inflammatory cytokines and growth factors as well as microvessel density (MVD) were determined at 14 days. Results showed more BMRMNCs were positive for CD34, CD45 and CD90. After transplantation, more viable cells were observed in the hippocampus of BMRMNCs treated rats. In addition, BMRMNCs transplantation significantly increased MVD, reduced pro-inflammatory cytokines and raised growth factors in the penumbra. However, the SDF-1 mRNA expression was comparable between BMRMNCs group and BMMNCs group. Our results indicate that BMRMNCs are likely to more effectively improve the local microenvironment to increase viable cells and elevate angiogenesis, exerting neuroprotective effects on cerebral ischemia in rats.

Total flavonoid aglycones extract in Radix scutellariae inhibits lung carcinoma and lung metastasis by affecting cell cycle and DNA synthesis.

ETHNOPHARMACOLOGICAL RELEVANCE: Radix Scutellariae (Scutellaria baicalensis Georgi, RS), a traditional herbal medicine commonly used to treat inflammation, hypertension, cardiovascular disease, bacterial and viral infections, is reported to treat lung cancer by supplements of modern medicine. The total flavonoid aglycones extract (TFAE) from RS is the most important composition for the pharmacodynamic effects. The present study was designed to evaluate the anti-lung tumor effect of TFAE on A549 cells and A549 cell nude mice xenografts. The aim of the study is to investigate the effect and mechanism of TFAE treating non-small cell lung cancer both in vitro and in vivo. MATERIALS AND METHODS: The anti-tumor activity of TFAE in vitro was investigated using the MTT assay. The changes of cell invasion and migration were detected by Transwell assay and tube formation experiments were used to detect the anti-angiogenic effect. The anti-tumor effects of TFAE in vivo were evaluated in A549 cell nude mice xenografts. The mechanism of TFAE was detected by flow cytometry technology, western blot assay and immuno-histochemistry assay. RESULTS: In vitro, TFAE inhibited the proliferation, invasion and migration of A549 cells in a dose- and time-dependent manner. In vivo, TFAE by oral administration at 100mg/kg for 30 days decreased the tumor volume and tumor weight in A549 cell xenograft by 25.5% with no statistical significance (P<0.05) compared to the cis-platinum positive control group (30.0%). The cell cycle and DNA synthesis experiment illustrated that TFAE could induce A549 cell cycle to arreste in S phase and DNA synthesis in A549 cells be inhibited, while TFAE had no influence on apoptosis of A549 cells. Western Blot assay demonstrated that the treatment of TFAE could make Cyclin D1 decrease and p53 increase both in vitro and in vivo. CONCLUSION: TFAE displayed the inhibition effects of non-small cell lung cancer both in vitro and in vivo and the underlying mechanism might be related to the increased p53 protein expression and decreased Cyclin D1 expression, leading to cell cycle arrested in S phase and the decrease of DNA synthesis.

Overexpression of 5-lipoxygenase increases the neuronal vulnerability of PC12 cells to Aß42.

5-Lipoxygenase (5-LOX), which is believed to be a major source of oxidative stress, participates in somatostatin-receptor transmembrane signaling in the central nervous system. We used the Tet-On inducible expression system in PC12 cells to obtain cell lines with reproducible, stable 5-LOX expression levels to study its function. Cell apoptosis rates induced by Aß(42) were determined using an apo-BrdDU kit. Lipid peroxide, antioxidant enzyme, and caspase-3 activities were evaluated with respective commercial kits. The expression of 5-LOX, bcl-2, and bax were detected by immunoblotting. A subclone of PC18 with Tet-On inducible expression of 5-LOX was selected from PC12 transfectants. Expression of 5-LOX had no significant inhibitory effect on the cell viability of the PC18 clone. In contrast, compared with the control group, the cell viability of clone PC18 was significantly reduced after the induction of 5-LOX during Aß exposure. The differences in cell viability before and after the induction of 5-LOX during Aß insult were significantly offset by AA861. Overexpression of 5-LOX only slightly improved the activities of superoxide dismutase (SOD). The levels of intracellular peroxides, SOD and caspase-3 activity, and Bax expression were significantly upregulated, and the levels of glutathione peroxidase and catalase were downregulated correspondingly in clone PC18 during Aß exposure. These results indicate that constitutive expression of 5-LOX is not detrimental per se, but overexpression of 5-LOX may become problematic during Aß exposure.

[Early morphological changes in the mouse testis induced by 5-fluorouracil].

OBJECTIVE: To propose a simple and practical method for preparing a mouse model of oligo-astheno-terato-spermia/azoospermia, and to offer a methodological suggestion for the studies on the related mechanism of spermatogenesis and evaluation of medication efficacy by observing the early changes of testis morphology after 5-fluorouracil treatment. METHODS: Mice were injected with a single dose of 5-flourouracil at 250 mg/kg via the tail vein, and their testes were harvested and paraffin sections prepared before and 3, 7, 11 and 14 d after the injection to be observed for the morphological changes by hematoxylin and eosin staining. RESULTS: The numbers of spermatocytes/spermatids were progressively reduced inside the testis seminiferous tubules of the mice at 3, 7 and 11 d after the 5-fluorouracil injection, and the tubule walls became thinner, which reached the nadir at 11 d, with evident swelling and crazing of the seminiferous tubules. At 14 d, the swelling almost disappeared and spermatocytes became repopulated, while the flaws still existed in the seminiferous tubules and no mature sperm were seen. CONCLUSION: One-dose injection of 5-fluorouracil via the tail vein might be a simple and effective method for preparing the animal model of reproductive function damage induced by chemotherapeutic medication.

[Interrelationship of abnormal family history in the first degree relatives and clinical phenotype of patients with polycystic ovary syndrome].

OBJECTIVE: To study the relationship of abnormal family history in the first degree relatives and the clinical phenotype of patients with polycystic ovary syndrome (PCOS). METHODS: Clinical data of first degree relatives of 139 women with PCOS were collected by questionnaires, including body mass index (BMI), waist hip ratio (WHR), and hursutism score. Follicle stimulating hormone (FSH), luteinizing hormone (LH), prolactin (PRL), testosterone (T), androstenedione (A), oral glucose tolerance test (OGTT) and insulin releasing test were measured. RESULTS: (1) Compared with patients with a negative family history of diabetes mellitus, for women with a positive family history, WHR (0.99 +/- 0.10 vs 0.79 +/- 0.08) and score of hirsutism (1.9 +/- 1.2 vs 1.8 +/- 1.2) were increased, the duration of menstruation was longer [(108 +/- 10) vs (92 +/- 19) days]; A [(11 +/- 6) vs (8 +/- 5) nmol/L], homeostasis model assessment of insulin resistance (HOMA-IR, 3.5 +/- 2.0 vs 2.7 +/- 1.6), area under curve (AUC) glucose [(836 +/- 245) vs (748 +/- 139) nmol.L(-1).min(-1)], AUC insulin [(9670 +/- 4582) vs (7330 +/- 4311) mIU.L(-1).min(-1)], fasting glucose [(5.0 +/- 1.1) vs (4.8 +/- 0.5) mmol/L] and fasting insulin [(15 +/- 8) vs (11 +/- 8) mIU/L] were increased, while early insulin secretion function index (DeltaI60/DeltaG60, 32 +/- 22 vs 52 +/- 30), insulin sensitive index (ISI, 0.019 +/- 0.011 vs 0.033 +/- 0.014) and disposition index (DI, 18 +/- 10 vs 30 +/- 22; P < 0.05) were decreased. (2) For women with a positive family history of menstrual disorder, WHR and score of hirsutism (0.99 +/- 0.09 vs 0.80 +/- 0.10 and 1.9 +/- 1.0 vs 1.6 +/- 1.1) were increased respectively, the duration of menstruation [(105 +/- 28) vs (84 +/- 31) days] was longer, HOMA-IR (3.6 +/- 2.4 vs 2.5 +/- 1.7) and fasting insulin level [(15 +/- 14) vs (12 +/- 11) mIU/L] were increased, while HOMA-beta (178 +/- 134 vs 207 +/- 175), ISI (0.017 +/- 0.009 vs 0.033 +/- 0.012) and DI (23 +/- 18 vs 28 +/- 19, P < 0.05) were decreased. (3) For women with a positive family history of premature balding, BMI and the score of hirsutism [(26 +/- 4) vs (23 +/- 5) kg/m(2) and 2.1 +/- 1.1 vs 1.7 +/- 1.3] were increased respectively, while DI (20 +/- 11 vs 30 +/- 23, P < 0.05) was decreased. (4) DeltaI60/DeltaG60 (34 +/- 27 vs 50 +/- 30) was decreased and fasting insulin [FINS, (13 +/- 10) vs (10 +/- 9) mIU/L] was increased in PCOS women with a family history of hypertension (P < 0.05). (5) For women with or without a family history of coronary heart disease, they did not have any difference in every parameter mentioned before. CONCLUSIONS: The family history of diabetes mellitus has the most effect on the clinical phenotype in women with PCOS. The family history of other diseases such as menstrual disorder, premature balding and hypertension play less significant roles. A family history of positive coronary heart disease does not affect the clinical phenotype of such patients.

Roles of single photo emission computed tomography-CT in detecting residual tumor from patients with thyroid carcinoma / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>As newly, functional, diagnostic measure-18F-deoxyglucose single photo emission computed tomography (18F-FDG SPECT-CT) has well sensitivity and specialty , it was proved availability in differentiating benign and malignant tumor. In this study, it was compared the efficiency of detecting residual tumor of thyroid cancer between 18F-FDG SPECT-CT and CT-TWIN.</p><p><b>METHODS</b>During May 2004 to August 2004, 56 patients with thyroid disease were divided into two groups, one group was 32 cases suffered thyroid tumor, another was 24 cases suffered improper operation. All cases must take 18F-FDG SPECT-CT and CT-TWIN.</p><p><b>RESULTS</b>Forty eight cases had been performed operation, and 8 cases waited and watched because they didn't be found residual tumor by 18F-FDG SPECT-CT and CT-TWIN. The results in detecting thyroid tumor showed that the same rate of diagnostic correction of SPECT-CT was 87.5% (28/32), and the Youden index of SPECT-CT was 0.667, higher than that of CT which was 87.5% (28/32) and 0.633 respectively. When the results two methods were accordant, the rate of diagnostic correction was 100% (24/24), and Youden index was 1, and the result had statistic signification. The rate of diagnostic correction of SPECT-CT in detecting residual tumor was 84.6%, and its Youden index was 0.675. The rate of diagnostic correction of CT in detecting residual tumor was 75.0%, and its Youden index was 0.492; and the result had statistics signification by being compared two different measures.</p><p><b>CONCLUSIONS</b>SPECT-CT and CT all had higher diagnostic efficacy, and they could promote the diagnostic efficacy when they have the same diagnosis in detecting thyroid tumor. The rate of detecting residual tumor of SPECT-CT was 81.3%, compared with that of CT the diagnostic efficacy has greater raise.</p>

Pathological character and treatment of epithelial-myoepithelial carcinoma of salivary gland / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To review and evaluate pathologic features and treatment of epithelial-myoepithelial.</p><p><b>METHODS</b>Retrospectively reviewed 14 cases' pathological and clinical materials of epithelial-myoepithelial carcinoma of salivary gland. Eight cases origine from parotid gland, 2 cases from hard palate, 3 cases from submandibular gland and 1 case from nasal cavity. Three cases were performed induction chemotherapy preoperation. One case had palliative radiotherapy. Thirteen cases were performed radical surgery and 6 cases had radiotherapy postoperation.</p><p><b>RESULTS</b>Tumor arisen mostly from parotid gland and neck lymph node metastasis rate was 14.28% (2/14). The survival rate was calculated with Kaplan-Meier method. The overall 3-, 5- and 10-year survival rate were 67.20%, 45.49% and 17.06%. Its histological characteristics were inner layer composed by adenoid cells and outer layer composed by myoepithelial cells. Immunohistochemical exam show cytokeratin, S-100 and actin reaction positive.</p><p><b>CONCLUSIONS</b>Epithelial-myoepithelial carcinoma easily develops recurrence. It is sensitivity to radiotherapy and chemotherapy to some extent. It is suitable to adopt surgical treatment as primary modality combined with other therapies.</p>

Value of elective neck dissection in patients with clinically stage I squamous cell carcinoma of the tongue / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To determine the value of elective neck dissection in patients with clinically stage I (cT1N0M0) squamous cell carcinoma of the tongue.</p><p><b>METHODS</b>This was a retrospective study of patients with surgical treatment between November 1984 and November 1999. A total of 130 patients were included in the study, all of whom received operation of the primary site, meanwhile, 99 of whom underwent elective neck dissection simultaneously including level I -III or level I -IV neck dissection in 20 patients and level I - V neck dissection in 79 patients. Results Among all these patients, the rate of occult metastasis to the neck were 12. 0%. Local failure rate in patients with only local treatment, level I II, II, III/IV neck dissection and level I - V neck dissection were 25. 8%, 15. 0% and 7. 6% respectively. There were significant difference in regional failure between patients with only local treatment and patients with elective neck dissection (P < 0.05). Also, no significant differences were noted in the survival rate between patients with only local treatment, elective neck dissection (level I -III or level I -IV) and level I -V neck dissection (P > 0.05).</p><p><b>CONCLUSIONS</b>Elective neck dissection significantly reduced regional control failure but was not able to reduce distant metastasis or increase the overall survival. A prospective randomized study is worthwhile to further evaluate the benefit of elective neck dissection in the treatment of clinically stage I squamous cell carcinoma of the tongue.</p>

Differentiated thyroid carcinoma in young people / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To investigate the factors that influence survival of the patients with differentiated thyroid carcinoma in young people and evaluate the efficiency of unilateral lobectomy plus isthmectomy with therapeutic cervical lymph node dissection and postoperative TSH (thyroid stimulating hormone) suppressive therapy.</p><p><b>METHODS</b>One hundred and thirty-one patients under 30 years old with differentiated thyroid carcinoma treated in this hospital (14 cases no more than and 117 cases more than 16 years) from Jan. 1st, 1985 to Dec. 31st, 1997 were retrospectively reviewed. One hundred and twenty-eight patients were received only surgery and TSH suppressive therapy, and 3 patients received chemotherapy or radiotherapy because of the progressive metastasis in necks or mediastina. A multivariate analysis was performed in these patients by the Cox proportional hazard model.</p><p><b>RESULTS</b>The mean follow-time (x +/- s) of all patients were (140.86 +/- 43.76) months, with range from 20 to 229 months; Ninety-eight patients followed more than 10 years. Ten patients died of thyroid cancer. The overall 10-year survival rate was 97.18%. The 10-year survival rate for patients < or = 16 years of age and > 16 years were 75.97% and 96.57% respectively (P = 0. 0006). The 10-year survival rate for women and men were 94.91% and 93.69% respectively (P = 0.5261). The 10-year survival rates of patients with papillary thyroid carcinoma and follicular thyroid carcinoma were 93.77% and 96. 55% respectively (P = 0.8137). For patients with tumor size of < or = 1 cm, 1-4 cm and >4 cm the survival rate was 100.0%, 96.40%, and 80.67% respectively (P = 0. 0589). The 10-year survival rates of patients with or without lymph node metastasis were 88.37% and 100. 0% respectively (P = 0.0313). For patients of with or without distant metastasis, The survival rate was 96.64% or 60.00% (P = 0.0000). The 10-year survival rates with or without recurrence were 86. 67% and 95.48% respectively (P = 0. 5681). Using multivariate analysis, risk factors that independently influence survival were distant metastasis, tumor size and age.</p><p><b>CONCLUSIONS</b>The distant metastasis, tumor size and age at diagnosis were the independent factors influencing survival significantly. The status of lymph node metastasis may have certain effect on the prognosis. Unilateral lobectomy plus isthmectomy with a therapeutic cervical lymph node dissection followed by postoperative TSH suppressive therapy is a favourable model to children and young adults with DTC without distant metastasis, but to the patients with distant metastasis, their prognosis of this therapy model is disappointing.</p>

Management option for cervical metastases in tongue squamous cell carcinoma with clinically N0 neck / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>This study was designed to explore the optimal management option for cervical metastases in tongue squamous cell carcinoma (SCC) with clinically N0 neck in order to avoid excessive or inadequate treatment in clinical practice.</p><p><b>METHODS</b>Clinical data of 327 cases of tongue SCC with cN0 neck were retrospectively analyzed. Neck control rates affected by different pathoclinical parameters were compared. Prognosis analysis and death analysis were also performed.</p><p><b>RESULTS</b>Overall 3-year survival was 69.7% (228/327), 3-year survival of neck recurrent group and non-recurrent group was 39.1% (25/64) and 77.2% (203/263), and 51.5% (51/99) of the death related to neck failure. Overall neck control rate was 80.4% (263/327); neck control rate of wait and watch group, level I neck dissection, level I + II neck dissection, supraomohyoid neck dissection, radical neck dissection, functional neck dissection, was 67.5% (27/40), 72.7% (24/33), 60.0% (15/25), 84.9% (45/55), 86.8% (131/151), 84.0% (21/25), respectively. Treatment modality and cervical lymph node involvement were independent factors for neck control.</p><p><b>CONCLUSIONS</b>Neck control is a key for prognosis of tongue SCC with cN0 neck. Supraomohyoid neck dissection is the first choice in management of cervical metastases in tongue SCC with cN0 neck, during which the suspected involved lymph nodes should be sent for frozen section to determine whether comprehensive neck dissection required. Multimodal metastasis and/or capsular spread are the indications for postoperative irradiation.</p>

Quantitative analysis of mitochondrial DNAs in macaque embryos reprogrammed by rabbit oocytes.

In cloned animals where somatic cell nuclei and oocytes are from the same or closely related species, the mitochondrial DNA (mtDNA) of the oocyte is dominantly inherited. However, in nuclear transfer (NT) embryos where nuclear donor and oocyte are from two distantly related species, the distribution of the mtDNA species is not known. Here we determined the levels of macaque and rabbit mtDNAs in macaque embryos reprogrammed by rabbit oocytes. Quantification using a real-time PCR method showed that both macaque and rabbit mtDNAs coexist in NT embryos at all preimplantation stages, with maternal mtDNA being dominant. Single NT embryos at the 1-cell stage immediately after fusion contained 2.6 x 10(4) copies of macaque mtDNA and 1.3 x 10(6) copies of rabbit mtDNA. Copy numbers of both mtDNA species did not change significantly from the 1-cell to the morula stages. In the single blastocyst, however, the number of rabbit mtDNA increased dramatically while macaque mtDNA decreased. The ratio of nuclear donor mtDNA to oocyte mtDNA dropped sharply from 2% at the 1-cell stage to 0.011% at the blastocyst stage. These results suggest that maternal mtDNA replicates after the morula stage.

Clinical research of sentinel node biopsy in oral tongue carcinoma / 中华口腔医学杂志

<p><b>OBJECTIVE</b>This study was designed to evaluate whether sentinel node (SN) biopsy can accurately assess the cervical lymph node status of oral tongue carcinoma, as well to research the best method and indications of SN biopsy.</p><p><b>METHODS</b>Preoperative lymphoscintigraphy with (99m)Tc-SC and intraoperative sentinel node mapping with methylene blue dye were administered on 20 cases of oral tongue carcinoma with cN(0) neck and 5 cases with cN(+) neck; routine pathological examination was used to assess the status of SNs. The results of routine pathological examination of cervical specimen were set as golden standard to assess the efficacy of SN biopsy in evaluating the cervical lymph node status.</p><p><b>RESULTS</b>53 SNs were detected in 24 cases out of the total 25 cases (96%), averaging 2.2 SNs per case. SNs were detected in all 20 cases with cN(0) neck, in which 4 cases with occult cervical metastasis were detected by SN diopsy, without false negative case found in the procedure. In 5 cases with cN(+) neck, SNs were detected in 4 cases. In 4 cases whose SNs were detected, there were 5 cN(+) necks, out of which SNs were detected in 4 cN(+) necks but failed to predicted the cervical lymph node status in 2 necks. However, SNs were detected in 2 out of the other 3 cN(0) necks, both of which were diagnosed as SN(+)pN(+).</p><p><b>CONCLUSIONS</b>Nuclear lymphoscintigraphy and blue dye mapping can be used to trace the SNs in cases with oral tongue carcinoma, with satisfactory detective rate. SN biopsy can accurately evaluate the cervical lymph node status in cases of oral tongue carcinoma with cN(0) neck. Whether it can be used to evaluate the lymph node status of the cN(0) neck in case with a contralateral cN(+) neck is worthy of further research.</p>

Embryonic stem cells generated by nuclear transfer of human somatic nuclei into rabbit oocytes.

To solve the problem of immune incompatibility, nuclear transplantation has been envisaged as a means to produce cells or tissues for human autologous transplantation. Here we have derived embryonic stem cells by the transfer of human somatic nuclei into rabbit oocytes. The number of blastocysts that developed from the fused nuclear transfer was comparable among nuclear donors at ages of 5, 42, 52 and 60 years, and nuclear transfer (NT) embryonic stem cells (ntES cells) were subsequently derived from each of the four age groups. These results suggest that human somatic nuclei can form ntES cells independent of the age of the donor. The derived ntES cells are human based on karyotype, isogenicity, in situ hybridization, PCR and immunocytochemistry with probes that distinguish between the various species. The ntES cells maintain the capability of sustained growth in an undifferentiated state, and form embryoid bodies, which, on further induction, give rise to cell types such as neuron and muscle, as well as mixed cell populations that express markers representative of all three germ layers. Thus, ntES cells derived from human somatic cells by NT to rabbit eggs retain phenotypes similar to those of conventional human ES cells, including the ability to undergo multilineage cellular differentiation.

Treatment of second primary malignant tumor induced by radiotherapy / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To study the diagnosis and treatment of a second primary malignant tumor induced by previous radiotherapy.</p><p><b>METHODS</b>From March 1970 to March 1997, 108 nasopharyngeal cancer (NPC) patients who developed a second primary malignant tumor induced by radiotherapy were treated. There were squamous carcinoma 43 (39.8%), sarcoma 26 (24.1%), malignant fibrous histiocytoma 14 (13.0%), adenoid cystic carcinoma 12 (11.1%), thyroid papillary adenocarcinoma 8 (7.4%) and malignant melanoma 5 (4.6%). Fifty patients underwent operation, 32 received radiotherapy, 18 received chemotherapy and 8 received operation combined with chemotherapy.</p><p><b>RESULTS</b>The 3- and 5-year tumor-free survival rates were 64.0% and 36.0% in the operation group. They were 34.4% and 18.8% in the radiotherapy group.</p><p><b>CONCLUSION</b>Surgery, if not contra-indicated, is the first choice for the second primary malignant tumor induced by radiotherapy. Aggressive treatment for these patients is, hence, indicated clinically.</p>