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.

Idiopathic recurrent facial palsy: Facial nerve decompression via middle cranial fossa approach.

OBJECTIVE: To introduce our experience of preventing further episodes of idiopathic recurrent facial palsy by facial nerve decompression via middle cranial fossa approach. METHODS: Twelve cases (surgery group) who had idiopathic recurrent facial palsy underwent facial nerve decompression via middle cranial fossa approach, and 6 cases (control group) who declined surgery accepted conservative treatment. Further episodes of facial palsy and final outcomes of facial nerve were recorded. Facial nerve function was assessed by House-Brackmann facial nerve grade system. RESULTS: Only 8.3% of the patients (one case) in the surgery group had further episodes of facial palsy on the surgical side, but up to 64.7% of the patients (4 cases) in the control group suffered further episodes (p<0.05). 11 patients (91.7%) in the surgery group recovered to Grade I or Grade II in contrast to 3 cases (50%) in the control group. CONCLUSIONS: Facial nerve decompression via middle cranial fossa approach was able to prevent further episodes of idiopathic recurrent facial palsy, and surgical decompression seemed to achieve better outcomes of facial nerve than conservative treatment.

[Analyses prognostic factors relevant to sudden sensorineural hearing loss].

OBJECTIVE: To investigate the prognostic factors relevant to sudden sensorineural hearing loss. METHODS: The internationally accepted standardized clinical research methods, unified design, and unified program were adopted to conduct the prospective clinical multi-center study. The sudden deafness patients between 18 to 65 years old, with the course of this disorder less than two weeks, and without any medical treatments were collected, and then, divided into four types according to the hearing curve: type A, acute sensorineural hearing loss in low tone frequencies; type B, acute sensorineural hearing loss in high tone frequencies; type C, acute sensorineural hearing loss in all frequencies; and type D, total deafness. The factors, in terms of age, gender, type of initial audiogram, time delay before the first visit, and severity of hearing loss, were included in the analyses. RESULTS: A total of 1 024 cases with single side sudden deafness were collected in the study from 33 hospitals in China from August 2007 to October 2011, inclusive of for 492 males (48.05%) and 532 females (51.95%). The average age was (41.2 ± 12.8) years old. There were 553 cases (54.00%) in left ear, and 471 cases (46.00%) in right ear. The curative effects of different types were shown as follows: the type in low tone frequencies had the highest rate of 90.73%, the type in all frequencies was 82.59%; the type of total deafness was 70.29%; and the type in high tone frequencies had the lowest rate of 65.96%. It had significant difference of the effective rate between different types (χ(2) = 231.58, P = 0.000). Age, time delay before first visit, and severity of initial hearing loss were significantly correlated with hearing improvement. CONCLUSIONS: Initial audiogram of SSNHL might predict hearing recovery. The young in age and a short time delay before starting treatment are positive prognostic factors for hearing recovery in SSNHL. The initial severity of hearing loss is negative prognostic factor of hearing recovery.

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.

Lipopolysaccharide-induced middle ear inflammation disrupts the cochlear intra-strial fluid-blood barrier through down-regulation of tight junction proteins.

Middle ear infection (or inflammation) is the most common pathological condition that causes fluid to accumulate in the middle ear, disrupting cochlear homeostasis. Lipopolysaccharide, a product of bacteriolysis, activates macrophages and causes release of inflammatory cytokines. Many studies have shown that lipopolysaccharides cause functional and structural changes in the inner ear similar to that of inflammation. However, it is specifically not known how lipopolysaccharides affect the blood-labyrinth barrier in the stria vascularis (intra-strial fluid-blood barrier), nor what the underlying mechanisms are. In this study, we used a cell culture-based in vitro model and animal-based in vivo model, combined with immunohistochemistry and a vascular leakage assay, to investigate lipopolysaccharide effects on the integrity of the mouse intra-strial fluid-blood barrier. Our results show lipopolysaccharide-induced local infection significantly affects intra-strial fluid-blood barrier component cells. Pericytes and perivascular-resident macrophage-like melanocytes are particularly affected, and the morphological and functional changes in these cells are accompanied by substantial changes in barrier integrity. Significant vascular leakage is found in the lipopolysaccharide treated-animals. Consistent with the findings from the in vivo animal model, the permeability of the endothelial cell monolayer to FITC-albumin was significantly higher in the lipopolysaccharide-treated monolayer than in an untreated endothelial cell monolayer. Further study has shown the lipopolysaccharide-induced inflammation to have a major effect on the expression of tight junctions in the blood barrier. Lipopolysaccharide was also shown to cause high frequency hearing loss, corroborated by previous reports from other laboratories. Our findings show lipopolysaccharide-evoked middle ear infection disrupts inner ear fluid balance, and its particular effects on the intra-strial fluid-blood barrier, essential for cochlear homeostasis. The barrier is degraded as the expression of tight junction-associated proteins such as zona occludens 1, occludin, and vascular endothelial cadherin are down-regulated.

Structural changes in thestrial blood-labyrinth barrier of aged C57BL/6 mice.

Tight control over cochlear blood flow (CoBF) and the blood-labyrinth barrier (BLB) in the striavascularis is critical for maintaining the ionic, fluid and energy balance necessary for hearing function. Inefficient CoBF and disruption of BLB integrity have long been considered major etiologic factors in a variety of hearing disorders. In this study, we investigate structural changes in the BLB of the striavascularis in age-graded C57BL/6 mice (1 to 21 months) with a focus on changes in two blood barrier accessory cells, namely pericytes (PCs) and perivascular-resident macrophage-like melanocytes (PVM/Ms). Decreased capillary density was detectable at 6 months, with significant capillary degeneration seen in 9- to 21-month-old mice. Reduced capillary density was highly correlated with lower numbers of PCs and PVM/Ms. "Drop-out" of PCs and "activation" of PVM/Ms were seen at 6 months, with drastic changes being observed by 21 months. With newly established in vitro three-dimensional cell-based co-culture models, we demonstrate that PCs and PVM/Ms are essential for maintaining cochlear vascular architecture and stability.

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.

[Clinical analysis of 26 patients with relapsing polychondritis].

OBJECTIVE: To discuss the clinical characteristics and early diagnosis and treatment of relapsing polychondritis(RP). METHOD: Twenty-six RP patients received the therapies of antibiotics, glucocorticosteroid, immunosuppressive agent,tracheotomy, tracheal sent implantation or non-invasive ventilation. RESULT: Twenty-six cases (88.5%) had auricle involvement, 20 cases (76.9%) had respiratory tract involvement, 18 cases (69.2%) had joints involvement, 10 cases (38.5%) had nose involvement, 6 cases (23.1%) had eyes involvement, 4 cases (15.4%) had cochlea and (or) vestibular involvement. Some patients with cardiac, vascular, kidney damage as well. One case died of severe pulmonary infection. One case died of respiratory failure. The symptoms of other 24 cases were stable or improved in different degrees. CONCLUSION: The clinical manifestations are diverse in RP patients, mainly in department of Otolaryngology, the prognoses of patients with airway involvement are worse, and it may improve the curative effect by an early diagnosis and a timely treatment.

Suppression of T-type Ca2+ channels inhibited human laryngeal squamous cell carcinoma cell proliferation running title: roles of T-type Ca2+ channels in LSCC cell proliferation.

BACKGROUND: Despite the tight correlation between T-type Ca2+ channels and a great variety of tumors, the roles of alpha1G subunit of T-type Ca2+ channels in laryngeal squamous cell carcinoma (LSCC) have not yet been investigated. METHODS: In the present study, we examined the expression of alpha1G subunit of T-type Ca2+ channel in human LSCC tissues and cell lines. One human laryngeal squamous cell carcinoma cell line, Hep-2, was also examined for T-type channels using voltage-clamp recordings. Cell proliferation assays were performed in the presence or absence of T-type channel blocker mibefradil and alpha1G subunit sepcific siRNA. The cell cycle was determined by flow cytometry. RESULTS: Our results indicated that the a1G subunit of T-type Ca2+ channel is highly expressed in human laryngeal squamous cell carcinoma tissues and cell lines. alpha1G siRNA significantly down-regulated the protein expression of the alpha1G subunit. Both alpha1G siRNA and mibefradil inhibited Hep-2 cell proliferation and arrested cell cycle progression. CONCLUSIONS: Together, these findings suggest a functional role of T-type channels in certain laryngeal carcinomas, and that inhibition of T-type channels reduces cell proliferation via cell cycle arrest, suggesting that the alpha1G subunit of T-type Ca2+ channel may be used as a therapeutic target for treating LSCC.

[Effect of H-uvulopalatopharyngoplasty on endothelium dependent flow-mediated dilation in patients with moderate to severe obstructive sleep apnea syndrome].

OBJECTIVE: This study evaluates the effects of H-uvulopalatopharyngoplasty (H-UPPP) on endothelial function by endothelium dependent flow-mediated dilation(FMD) in patients with moderate to severe obstructive sleep apnea syndrome (OSAHS). METHOD: Forty-five healthy subjects and 49 patients with moderately to severe OSAHS who desired H-UPPP were prospectively enrolled. FMD was measured by high-resolution B-mode ultrasonography. All subjects participated in sleep studies. These studies were repeated 6 months after H-UPPP in patients with moderately to severe OSAHS. RESULT: There was significant difference in FMD between patients with moderately to severe OSAHS and healthy subjects at baseline (6.5 +/- 2.1)% vs (11.2 +/- 3.1)% (P < 0.01). FMD increased from (6.5 +/- 2.1)% to (9.7 +/- 2.7)% 6 months after H-UPPP in patients with moderately to severe OSAHS (P < 0.01). CONCLUSION: FMD is decreased in patients with moderately to severe OSAHS, H-UPPP treatment of these patients leads to improvement of FMD.

[The observation on curative effect of 90 cases of sudden hearing loss].

OBJECTIVE: To evaluate the efficiency between two treatments of sudden hearing loss. METHOD: All patients were divided into two groups randomly, basic drug group was treated with ganglioside and vinpocetine injection, combined therapy group was treated with intratympanic dexamethasone and what was used in basic drug group. RESULT: The effective rate of combined therapy group (73.53%) was significantly higher than that of basic drug group (37.78%) (P < 0.05). CONCLUSION: The comprehensive therapy of intratympanic dexamethasone injection, ganglioside and vinpocetine injection have excellent efficiency for sudden hearing loss.

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.

[The effects of the plastic operation on upper respiratory tract at one-stage for patients with OSAHS through the assessment of 3-D reconstruction of CT images].

OBJECTIVE: The method of three-dimensional reconstruction of CT images is adopted to evaluate the effects of the plastic operation on upper respiratory tract at one-stage (stage one) for patients with multi-level obstructive sleep apnea hypopnea syndrome. METHOD: Polysomnography is used to diagnose patients with OSAHS. The obstruction parts of patients with OSAHS are determined by CT and endoscopy examinations in the upper air way. To reconstruct the tri-dimensional cavity of upper respiratory tract from data, the CT examinations on the sinuses and nasopharynx are rescanned to obtain the CT data after surgery. The changed situations before and after surgery are compared, and the narrow parts of the upper respiratory tract can be identified and measured intuitively. RESULT: Symptoms and PSG test results show that the effects of the surgical results are very good. and the cross sections of upper respiratory tract narrow parts were enlarged by more than 100%. Hydromechanical evidence approved the same results. CONCLUSION: Calculating the increment of the cross sections of the upper respiratory tract through tri-dimensional reconstruction can not only find the preoperative stenosis, but also assess the surgical effects objectively. Since the body of endoscope has a certain volume, surface anesthesia is needed to enter the nasal cavity and nasopharyngeal which will impact the objectivity of the assessment of the upper respiratory tract stenosis.

[Computerized three-dimensional reconstruction of cochlear microstructures].

OBJECTIVE: To explore an useful and practical method for three-dimensional reconstruction of cochlear microstructure. METHOD: A digital image dataset of magnified forty times of human cochlear tissue can be get from the serial sections of transverse cedukol images of male cadavers temporal bone. Photoshop 6.0 and Amira 3.0 software were used to deal with the digital image dataset, such as image-connecting and image inner-location and three-dimensional reconstruction of cochlear and scala media. RESULT: The result of three-dimensional reconstruction of the cedukol images represents the anatomy of human cochlear and scala media transparently and objectively. CONCLUSION: The three-dimensional reconstruction technique used in the study is practical for three-dimensional of inner ear microstructure.

[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.