Complement components of nerve regeneration conditioned fluid influence the microenvironment of nerve regeneration.

Nerve regeneration conditioned fluid is secreted by nerve stumps inside a nerve regeneration chamber. A better understanding of the proteinogram of nerve regeneration conditioned fluid can provide evidence for studying the role of the microenvironment in peripheral nerve regeneration. In this study, we used cylindrical silicone tubes as the nerve regeneration chamber model for the repair of injured rat sciatic nerve. Isobaric tags for relative and absolute quantitation proteomics technology and western blot analysis confirmed that there were more than 10 complement components (complement factor I, C1q-A, C1q-B, C2, C3, C4, C5, C7, C8ß and complement factor D) in the nerve regeneration conditioned fluid and each varied at different time points. These findings suggest that all these complement components have a functional role in nerve regeneration.

Effects of different combinations of growth factors on the differentiation of neural stem cells into hair-like cells.

The differentiation of neural stem cells (NSCs) is influenced by a variety of factors. Therefore, it is important to explore the best external conditions that will induce NSCs to differentiate into hair cells. In this study, we investigated the best in vitro conditions for differentiation of NSCs derived from the hippocampus of newborn guinea pigs into hair-like cells. NSCs were separated and induced in different combinations of growth factors-in a control group and 7 combinations. Myosin VIIa-positive cells were detected to compare the effects of various combinations of growth factors on the differentiation of NSCs into hair-like cells. NSCs were differentiated into hair-like cells in all groups, but cell growth was best in the basic fibroblast growth factor (bFGF) + epidermal growth factor (EGF) group and the bFGF + EGF + brain-derived neurotrophic factor (BDNF) group. The rates of myosin VIIa-positive cells in the 8 groups studied ranged from 13.53 to 22.71%, but the results in the bFGF+EGF and bFGF+EGF+BDNF groups had statistical significance compared with other groups (p < 0.05). While bFGF, EGF, and BDNF all can induce the differentiation of NSCs into hair-like cells, the synergies of bFGF+EGF and bFGF+EGF+BDNF are the best.

Construction and identification of multiple genes Co silence of plasmid shRNA.

Objective To construct and identify the eukaryotic vector expressing shRNA (Plasmid-1), which expressed the VEGF, C-myc, Survivin and hTERT gene at the same time. To detect its interference effects on the nasopharyngeal carcinoma cell line (CNE-2Z) compared with single gene plasmid VEFG (Plasmid-2). Methods According to the sequence of VEGF, C-myc, Survivin and hTERT gene, we designed 2 oligonucleotide sequences and synthesized a complementary DNA chain, then inserted it into the eukaryotic vector expressing pGenesil 1. The cell proliferation activity was detected by MTT method. The interference efficacy on human nasopharyngeal carcinoma cell line (CNE-2Z) in the level of mRNA and protein were detected by RT-PCR and Western-bolt. The inhibitory effect of plasmid on tumor in nude mice was also observed in vivo. Results The restriction enzyme digestion and sequencing technologies confirmed the construction of recombinant eukaryotic vector expressing was correct. The plasmid was transfected into CNE-2Z cells, green fluorescence can be seen clearly in the single gene and multi gene transfected cells under fluorescent microscope. MTT showed that the proliferation of cell was inhibited, the invasive ability was decreased in vitro, and the inhibition effects of single gene plasmid on the growth and proliferation of cells were lower than multi gene. Real-time-PCR and Western-bolt confirmed that the expression of target gene was decreased in the level of mRNA and protein, and the interference effect of multi gene was better than the single gene. The nude mice experiment showed that the interference effect of shRNA plasmid on the growth of tumor cell was better than single gene plasmid Conclusion We constructed a shRNA plasmid encoded four different genes successfully. After transfected with nasopharyngeal carcinoma cells, it can interfere the expression of VEGF, C-myc, Survivin and hTERT gene at the same time. And the interference effect was better than silence VEGF alone. Out results may provide experimental basis for multi gene therapy in the treatment of nasopharyngeal carcinoma.

Internal Mammary Artery Perforator-Supercharged Prefabricated Cervicothoracic Flap for Face and Neck Reconstruction.

BACKGROUND: Prefabricated flap is an important technique to reconstruct massive face and neck skin defects. But its vascularization remains unpredictable and often leads to abnormal blood supply of the harvested flap, even necrosis. Flap supercharging and turbo supercharging techniques are effectively used to improve flap blood supply. However, few studies have been reported on the application of these techniques in prefabricated induced expanded flaps. METHODS: From March 2008 to September 2012, 13 patients who have face and neck soft tissue defects were treated with prefabricated cervicothoracic flap. To overcome insufficient blood supply, 5 of them received additional microvascular augmentation in which the second or third perforator of the internal mammary artery (IMAP) and its venae comitantes were anastomosed to facial or superficial temporal vessels, contrary to the remaining 8 patients. The following results were compared: flap viability, hospital stay, complications, frequency of dressing change, reoperation rate, and remaining scars. RESULTS: No flap necrosis was observed in patients who received the supercharging procedure. By contrast, of the 8 patients who were not treated with supercharging technique, various degrees of flap necrosis occurred in 3 patients, 2 of whom received secondary operations. The frequency of dressing changes, the hospital stay, and hospital cost were reduced. Postoperative view showed better aesthetic restoration. CONCLUSIONS: The IMAP-supercharged cervicothoracic flap technique offers a reliable method for massive face and neck reconstruction. We recommended that the IMAP should always be preserved in the flap as a saving option for potential flap congestion or arterial insufficiency.

Ethical issues in Chinese aesthetic surgery.

Guided by the medical ethics principles of "four principles plus scope," Chinese aesthetic medical practitioners have proposed some extremely valuable ethical principles combined with the construction of aesthetic medicine and the requirements of clinical practice such as the principle of general nonmaleficence, the principle of local minimal invasiveness, the principle of informed consent, and the principle of respect and confidentiality. Chinese aesthetic surgical ethics provide valuable guidance for the practice of aesthetic medicine. Adherence to the ethics of Chinese aesthetic surgery provides an essential guide for the practice of aesthetic medicine in China. These principles protect both the medical practitioner and the patient, helping them to avoid unnecessary risks and disputes and ultimately promoting the sustainable development of aesthetic medicine.

Effect of Han-uvulopalatopharyngoplasty on flow-mediated dilatation in patients with moderate or severe obstructive sleep apnea syndrome.

CONCLUSION: Flow-mediated dilatation (FMD) is decreased in patients with moderate or severe obstructive sleep apnea syndrome (OSAS), and Han-uvulopalatopharyngoplasty (H-UPPP) can improve FMD. OBJECTIVE: To evaluate FMD in patients with moderate or severe OSAS and observe the effect of H-UPPP on FMD in these patients. METHODS: Forty-nine patients who were first diagnosed with moderate or severe OSAS by polysomnography (PSG) and had no other diseases served as the experimental group, and 35 individuals with normal PSG as the control group. FMD was measured with high-resolution B-mode ultrasonography in the two groups. PSG and FMD were again performed in the experimental group 6 months after H-UPPP. RESULTS: FMD was significantly lower in the experimental group than in the control group (6.5 ± 2.1% vs 11.2 ± 2.9%, p < 0.01). FMD was significantly improved 6 months after H-UPPP compared with preoperative FMD (9.7 ± 2.7% vs 6.5 ± 2.1%, p < 0.01).

Cancer stem-like side population cells in the human nasopharyngeal carcinoma cell line cne-2 possess epithelial mesenchymal transition properties in association with metastasis.

It has been recently reported that side population (SP) cells in nasopharyngeal carcinoma (NPC) cell lines display characteristics of cancer stem-like cells. However, the biological behavior and the significance of these cells for NPC progression remain unclear. In this study, we isolated SP cells from the NPC cell line CNE-2 by flow cytometry and investigated their biological characteristics. We discovered that SP cells had stronger colony forming abilities compared to the non-side population (NSP) cells, and observed that some SP cells looked more like the shape of mesenchymal cells when cultured in the common polyHEMA-coated flask. When checked by quantitative real-time PCR, the SP cells expressed higher levels of stemness-related genes Oct4, Sox2 and Nanog, and mesenchymal cell-related genes N-cadherin, vimentin and Snail, while they expressed lower levels of the epithelial cell-related gene, E-cadherin. Western blot and immunofluorescence staining methods further verified that SP cells expressed higher vimentin and expressed lower E-cadherin levels. Finally, Transwell invasion assay results indicated that the SP cells had higher invasive potential compared to NSP cells. Collectively, our data reveal that SP cells in the CNE-2 cell line not only possess the properties of cancer stem cells, but also have more mesenchymal cell characteristics which are associated with epithelial mesenchymal transition (EMT) and cancer cell invasion and metastasis. These findings are helpful for developing novel targets for effective clinical treatment of NPC.

[Effects of RNA interference targeting four different genes on the growth and proliferation of nasopharyngeal carcinoma CNE-2Z cells].

OBJECTIVE: To explore the effects of RNA interference (RNAi) targeting four different genes (VEGF, c-myc, survivin, hTERT) on the growth and proliferation of nasopharyngeal carcinoma (NPC) CNE-2Z cells. METHODS: Plasmid-1 targeted all four genes, plasmid-2, 3, 4 and 5 targeted VEGF, c-myc, survivin and hTERT respectively. These plasmids were transfected separately into human NPC CNE-2Z cells and xenograft tumors in nude mice. The expressions of plasmids in NPC CNE-2Z cells and xenograft tumors were observed. Cell proliferation was detected with MTT assay. The inhibitory effects on target genes were evaluated with RT-PCR and Western blot respectively. The effects of the plasmids on the biological behavior of CNE-2Z cells were observed with Transwell invasion chamber model. Apoptosis was determined with flow cytometer. The inhibitory effect of the plasmids on xenograft tumors were observed in nude mice. RESULTS: CNE-2Z cell proliferation was significantly inhibited and in vitro invasion ability was significantly decreased in the plasmid-1 group compared with those in the plasmid 2 - 5 groups (all P < 0.05). mRNA and protein expressions of all four genes decreased in the plasmid-1 group. The apoptosis rate in the plasmid-1 group was higher than that in the plasmid 2 - 5 groups (all P < 0.05). Growth of xenograft tumors in nude mice were significantly inhibited in the plasmid 1 - 5 groups, particularly in the plasmid-1 group. CONCLUSION: RNA interference targeting multiple genes can effectively inhibit NPC proliferation and induce apoptosis, which provides an experiment basis for NPC gene therapy.

Stem cell and peripheral nerve injury and repair.

Peripheral motor nerve injuries are a significant source of morbidity. Neural stem cells (NSCs), a group of relatively primitive cells, possess self-renewal ability and multidifferentiation potential. NSCs may be successfully separated from the human embryo and central nervous system (CNS) and differentiated into mature neurons and gliacytes by in vitro induction or transplantation into the body and may be differentiated into Schwann-like cells under specific conditions. It has been demonstrated that the ability of peripheral nerves to regenerate is mainly attributable to Schwann cells. NSC transplantation can promote peripheral nerve regeneration and provide a new means for treatment of peripheral nerve injury. In recent years, the study of NSCs has become a focus of many laboratories, but the biological characteristics and differentiation regulation mechanisms are not fully clear. In this article, we provide a brief review of NSC characteristics, cultivation, oriented differentiation, and clinical application.

[Effect of silencing VEGF, c-myc, survivin and hTERT by multiple shRNA expression vectors on the xenografted human nasopharyngeal carcinoma in nude mice].

OBJECTIVE: To investigate the effects of multiple short hairpin (shRNA) expression vectors, targeting VEGF, c-myc, survivin and hTERT, genes on the xenografted human nasopharyngeal carcinoma (CNE-2Z) in nude mice. METHODS: The shRNA expression vectors were constructed and subsequently transfected by direct injections into the tumors formed by CNE-2Z cells implanted in nude mice. The expressions of the targeted genes in tumor tissues and the apoptosis of tumor cells were evaluated. RESULTS: NPC CNE-2Z cells were successfully inoculated and subcutaneous tumor was formed in all nude mice. Under fluorescence microscope, tumor tissues showed the expression of each vector with green fluorescence. The expression of multiple shRNAs led to the decreases in the expressions of VEGF, c-myc, survivin, hTERT mRNA and proteins. Multi-gene silencing was better than single gene silencing in inducing the apoptosis of tumor cells. Tumor growth curves showed that the tumors treated with the shRNAs, including VEGF, c-myc, survivin, hTERT or the combination of 4 shRNAs, grower slowly obviously compared with control tumors. Inhibited rates of tumor growth by VEGF-, c-myc-, survivin- and hTERT-shRNA were 46.2%, 48.5%, 51.9% and 46.8% respectively. The combined application of 4 shRNA produced the more significant inhibitory rate (82.4%) than single shRNA application. CONCLUSIONS: The application of vector-based RNAi targeting multiple genes is a promising therapeutic modality in the gene therapy of nasopharyngeal carcinoma and multi-gene silencing is a new strategy for tumor therapy.

[Clinical features of parathyroid space-occupying lesions].

OBJECTIVE: To investigate the clinical manifestation, diagnosis, treatment of parathyroid occupying lesions. METHODS: The clinical data of 42 patients with parathyroid occupying lesions were retrospectively analyzed, including the clinical symptoms and signs, laboratory results, pathologic and imaging results and treatment. RESULTS: The number of males and females were 8 and 34, with females: males ratio being 1:5.25. The median age was 39 years. There were 2 cases of parathyroid cancer, 29 cases of parathyroid adenoma, 11 cases of parathyroid cysts in this study. The symptoms were as follows: 40 cases of neck lump, 34 cases of osteoporosis/osteitis fibrosa cystica, 29 cases of urinary symptom, 7 cases of voice hoarseness, 4 cases of peptic ulcer, 3 cases of dyspnoea and dysphagia, 3 cases of thoracic cavity lump, 2 cases of enhanced amylase activity. Serum calcium ion level and serum parathyroid hormone (PTH) level were examined qualitatively before operation. Ultrasonography, ECT-99mTc-MIBI, CT, MRI were used in diagnosing and locating parathyroid occupying lesions before operation. Twenty nine cases of parathyroid adenoma were treated with operation, 28 patients achieved complete remission, 1 suffered relapse after 23 months postoperative follow up. Eleven cases of parathyroid cysts were treated with operation and the outcome was no recurrence. Two cases of parathyroid cancer survived with out recurrence during follow up for 28 months and 50 months after operation. CONCLUSIONS: Examination of serum calcium and PTH level together with ultrasonography, ECT-99mTc-MIBI, CT, MRI is helpful to diagnose parathyroid occupying lesions. Surgery should be done as primary treatment. Tumor resection can be performed for parathyroid cysts, intraoperative exploration of bilateral neck is indicated for parathyroid adenoma, and a radical resection should be performed primarily for the parathyroid cancer.

Application of neural stem cells in tissue-engineered artificial nerve.

OBJECTIVE: To observe the curative effect of neural stem cells (NSCs), which are used in tissue-engineered artificial nerve, on repairing rabbit 10-mm facial nerve defects. METHODS: Thirty-six Oryctolagus cuniculi were randomly divided into three groups (each group with 12 Oryctolagus cuniculi). In group A, chitosan conduit, collagen protein sponge, nerve growth factor (NGF), and NSCs were used. In group B, chitosan conduit, collagen sponge, and NGF were used. In group C, nerve autograft was performed. Electrophysiologic detection, histologic observation, and BrdU and S100 immunohistochemical examination were performed 12 weeks after operation. RESULTS: All observation items in group A were better than those in group B (P < 0.01), and there were no significant differences between group A and group C (P > 0.05). CONCLUSION: NSCs may be served as seed cells of peripheral nerve tissue engineering and be used in artificial nerve to repair facial nerve defects.

[Effects of sodium butyrate on growth, apoptosis and telomerase activity in Hep-2 cells].

OBJECTIVE: To study the effects of sodium butyrate (SB) on growth, apoptosis and telomerase activity in Hep-2 cells. METHODS: Growth inhibition effect of SB on Hep-2 cells was assessed by methyl thiazolyl tetrazolium (MTT) assay. Morphological alterations were observed by electronic microscope. Cell apoptosis was confirmed by terminal deoxynucleotidyl transferase-mediated dUTP nick-end-labeling (TUNEL) method, DNA fragmentation and flow cytometry (FCM). Cell cycle was analyzed by FCM. Telomerase activity was examined by telomeric repeat amplification protocol (TRAP)-silver staining. The expression status of telomerase subunits was analyzed by reverse transcription-polymerase chain reaction (RT-PCR). RESULTS: A time-and dose-dependent inhibition was detected in cells treated with SB. Typical morphological changes of apoptotic cells were observed under electronic microscopy. The characteristic DNA fragmentation of apoptotic cells was detected by agarose gel electrophoresis. Apoptosis and the changes of cell cycle were confirmed by TUNEL method and FCM. The apoptosis indexes of the cells before treatment and at 72 h after SB (2.5 mmol/L) treatment were 2.27 +/- 1.18 and 33.50 +/- 2.75 respectively, the apoptosis rates were 2. 86% and 31. 28% respectively, the proportion of the cells at G0/G1 stage were 50.38% and 70.88% respectively, the proportion of the cells at S stage were 27.40% and 8.20% respectively, and the proliferation indexes of the cells were 49.62% and 29.12% respectively. Telomerase activity and expression level of human telomerase reverse transcriptase (hTERT), the key subunit of telomerase, decreased after SB treatment. No significant changes were observed in the expression of human telomerase RNA (hTR) and human telomerase associated protein (hTP1), the other two subunit of telomerase. CONCLUSION: SB could inhibit growth of Hep-2 cells and induce apoptosis in the cells, and inhibit telomerase activity by decrease expression level of hTERT.

[Observation of auditory brainstem response and distortion product otoacoustic emission on the animal model of autoimmune auditory neuropathy].

OBJECTIVE: To set up an animal model of autoimmune auditory neuropathy and to observe the auditory brainstem response (ABR) and distortion product otoacoustic emission (DPOAE) in guinea pigs. METHODS: The spiral ganglion and the cochlear nerve were obtained and purified by electrophoresis from 250 normal guinea pigs. The purified cochlear nerve antigen was mixed with an equal volume of complete Freunds adjuvant for immunization. Seventy guinea pigs were divided into three groups: experiment group (50 guinea pigs), control group (10 guinea pigs), normal group (10 guinea pigs). ABR, DPOAE, serum IgG levels, and morphological changes of spiral ganglion cells and the cochlear nucleus were observed. The protein expressions of the antigen were examined by immunohistochemistry and the super-structure of the auditory nerve were observed. RESULTS: The threshold of ABR response increased ranged from 10 to 25 dB in 32% (32/100 ears) of the guinea pigs. The peak latencies of waves I , III and the interpeak latency I approximately III were prolonged in the hearing loss group of guinea pigs. Prolonged peak latency of wave III was noted in hearing loss group at 2 and 3 weeks post immunization and slowly decreased to normal peak latency. The amplitude of DPOAE was no difference in the guinea pigs. The levels of serum IgG increased significantly compared with those of the control group. Inflammatory cell infiltration was observed in the cochlear nerve and the number of spiral ganglion cells detected. On the contrary, inflammatory cell infiltration was not observed in the cochlear nucleus. The cell densities and the across-sectional areas of neurons in anteroventral cochlear nucleus and posteroventral cochlear nucleus were no difference in the guinea pigs. The antigen protein distributed strictly in cochlear nerve and the spiral ganglion. Some demyelinated areas in cochlear nerve was observed in this group. The threshold of ABR response in 68% guinea pigs (68/100 ears) did not increase. The data of DPOAE and the serum IgG levels show no difference compared with the control group. There were not pathological observation in spiral ganglion cells, cochlear nucleus and cochlear nerve. CONCLUSION: An animal model of autoimmune auditory neuropathy has been set up successfully and the character of the ABR and DPOAE was observed.

Dexamethasone pharmacokinetics in Guinea pig inner ear perilymph.

AIM: To study the dexamethasone pharmacokinetics in the inner ear perilymph of guinea pigs using high-pressure liquid chromatography. METHODS: Sixty-five guinea pigs were divided into three groups. In the first group, the drug application protocol used an intra-abdominal dose of 0.5% dexamethasone 4 mg x kg(-1). In the second group, an intratympanic application dose of 0.5% dexamethasone 150 microl was used. The third group was the control group. The concentrations of dexamethasone in inner ear perilymph were determined by high-pressure liquid chromatography. RESULTS: The perilymph concentration-time curves of dexamethasone conformed to a one-compartment open model after an intra-abdominal application. The Cmax was 0.927 +/- 0.008 mg x l(-1), the Tmax 1.47 +/- 0.04 h, the T(1/2K) 2.92 +/- 0.056 h, the AUC 5.533 +/- 0.05 mg x h x l(-1), the T(1/2Ka) 0.47 +/- 0.024 h. After an intratympanic application, the perilymph concentration-time curves of dexamethasone also conformed to a one-compartment open model. The Cmax was 0.201 +/- 0.006 mg x l(-1), the Tmax 0.117 +/- 0.06 h, the AUC 0.868 +/- 0.004 mg x h x l(-1), the T(1/2K) 2.918 +/- 0.089 h, the T(1/2Ka) 0.161 +/- 0.009 h. Compared to the intra-abdominal application, the intratympanic application resulted in similar levels of inner ear perilymph drug concentrations in 30 min. CONCLUSION: Dexamethasone can penetrate the blood-labyrinthine barrier after intra-abdominal application. Dexamethasone can enter into perilymph after intratympanic application. Under the condition of the study, the intratympanic application resulted in a similar level of inner ear perilymph drug concentrations compared to the intra-abdominal application in 30 min.

[Structure and function influence of cochlear after dexamethasone intratympanic application].

OBJECTIVE: To study the changes of the structure and function of cochlear after dexamethasone intratympanic application. METHODS: Forty-five guinea pigs were divided into three groups as normal group, 0.9% sodium chloride injection group and 0.5% dexamethasone group. By using the auditory brainstem response (ABR), the superoxide dismutase (SOD) activity, the nitric oxide (NO) value and cochlear surface preparation technique, the changes of structure and function of cochlear after intratympanic Dexamethasone application has been investigated in this study. RESULTS: There was not significant difference among three groups in ABR thresholds of wave III (F = 0.5, P = 0.5). There was not significant difference among three groups in SOD activity (F = 2.45, P = 0.96). There was not significant difference among three groups in NO value (F = 3.1, P = 0.3). There was not significant difference among three groups in necrotic values of hair cells on the cochlear surface specimens in 15 mm from basal turn (F = 0.93, P = 0.45). CONCLUSION: There is no change of cochlear construction and function after intratympanic Dexamethasone application.

Functional remobilization evaluation of the paralyzed vocal cord by end-to-side neurorrhaphy in rats.

OBJECTIVE: To investigate the value of end-to-side neurorrhaphy to treat vocal cord paralysis. STUDY DESIGN: A prospective study evaluating the effects of end-to-side neurorrhaphy to treat vocal cord paralysis by means of fiberoptic laryngoscopy and nerve electromyography. METHODS: Thirty Sprague-Dawley rats were divided into experimental group 1, experimental group 2, and a control group randomly. Right recurrent laryngeal nerve (RLN) was incised, and the distal end of the RLN was anastomosed to the right phrenic nerve by end-to-side neurorrhaphy in experimental group 1 or by end-to-end nerve anastomosis in experimental group 2, respectively. The adductor nerve branch of the right RLN was incised and anastomosed to the proximal end of the right ansa cervicalis nerve by end-to-end nerve anastomosis. Fiberoptic laryngoscopy and nerve electromyography were used to examine the vocal cord movement and nerve regeneration. RESULTS: Three months after operation, this effect of end-to-side neurorrhaphy created a significant difference compared with the end-to-end nerve anastomosis (P < .05). The end-to-side neurorrhaphy did not lead to vocal cord movement compared with end-to-end nerve anastomosis. CONCLUSION: Vocal cord paralysis cannot be treated by this microsurgical technique.

[Microsatellite instability and loss of heterozygosity of fragile histidine triad gene in laryngeal squamous cell carcinoma].

OBJECTIVE: To investigate the loss of heterozygosity (LOH) and microsatellite instability (MSI) of fragile histidine triad (FHIT) gene in laryngeal squamous cell carcinoma (LSCC). METHODS: Using polymerase chain reaction-simple sequence length polymorphism (SSLP) -silver staining technique, 41 samples of LSCC were detected for LOH and MSI with microsatellite sites D3S1234 and D3S1300. RESULTS: LOH was found in 44.4% (16/36) and 36.4% (12/33) of informative cases at D3S1234 and D3S1300 respectively, while MSI at the two loci was found in 19.4% (7/36) and 24.2% (8/33) of informative cases respectively. In 41 cases of LSCC, 38 cases were informative at either locus. 52.6% (20/38) of cases had LOH while 28.9% (11/38) had MSI. The rate of LOH was related to the tumor TNM stage, pathological grade, lymph node metastasis and tumor relapse (P < 0.05). The rate of MSI was related to the tumor lymph node metastasis (P < 0.05 ). CONCLUSION: The frequencies of LOH and MSI in the two microsatellite sites, D3S1234 and D3S1300, of FHIT gene might play a role in carcinogenesis and development of LSCC and might provide some new approaches for early gene detection for LSCC.

[Distribution of basic fibroblast growth factor in inner ear of guinea pigs injected by different drug-approaches].

OBJECTIVE: To investigate the approach of basic fibroblast growth factor(bFGF) entering inner ear, as well as its the protective mechanism to inner ear and nerve tissue in pathological situation. METHODS: 125I-bFGF was injected into guinea pigs body via the lateral ventricle and muscle under physical situation as well as pathological situation. Then the per minute gamma-radioactive in blood, liver, thyroid gland, brain, cochlear and perilymph fluid was counted, and the distribution and metabolism of bFGF in the inner ear and autoradiography of the cochlea were also observed. RESULTS: Gamma-radioactive cpm of blood and liver increased significantly, while it did not change in brain, cochlea and perilymph after 125I-bFGF intramuscular injections. Gamma-radioactive cpm in blood, liver, brain, perilymph and cochlea had increased and autoradiography granules was found in the cochlea in 30 min after 125I-bFGF injected into CSF. In brain, perilymph and cochlea, a maximal value of gamma-radioactive cpm was obtained between 2 h and 4 h, while that in 8 h decreased significantly. Autoradiography granules still were seen in 8 h. gamma-radioactive cpm in 12 h was still higher than that in control group, but autoradiography granules can't be seen. The result in 24 h was similar to that in control group. The time course of cpm in the blood, cochlea and perilymph always parallel changed. CONCLUSIONS: bFGF has some difficulties in getting across blood-labyrinth barrier (BLB) and blood-brain barrier (BBB) under physical and pathological situation, such as acute anoxia, aminoglycoside-induced deafness. bFGF can reach inner ear, perilymph fluid, brain tissue and blood rapidly when it is injected into CSF and excreted slowly in those tissues. Permeability of BBB and BLB to bFGF is similar and has orientation.