Preparation and performance of a pH-sensitive cisplatin-loaded magnetic nanomedicine that targets tumor cells via folate receptor mediation.

The present study aimed to prepare cisplatin (CDDP)-loaded magnetic nanoparticles (MNPs), which target folate receptors via a pH-sensitive release system (FA­PEG­NH­N=MNPs­CDDP). This is of interest for the development of intelligent drug delivery systems that target tumors of the head and neck. The chemical coprecipitation method was used to prepare ferroferric oxide MNPs. These were modified with aldehyde sodium alginate complexed with the chemotherapeutic agent, CDDP on the surface of the nanoparticles. Double hydrazine­poly(ethylene glycol; PEG) was also prepared by attaching the carboxyl group of hydrazine­folate on one side of the double hydrazine­PEG, obtaining folate­hydrazine­PEG­diazenyl. This binds the aldehyde group of sodium alginic acid on the MNP to enclose CDDP, in order that it is sequestered within the carrier. This method obtained a pH­sensitive, FA­modified CDDP­loaded MNP (FA­PEG­NH­N=MNPs­CDDP), which acts as an intelligent tumor targeting drug delivery system. The mean size of the MNPs was ~10.2±1.5 nm, the mean hydrodynamic diameter detected by laser particle sizing instruments was 176.6±1.1 nm, and the ζ­potential was ­20.91±1.76 mV. The CDDP content was 0.773 mg/ml, the iron content was ~1.908 mg/ml and the maximum saturation magnetization was 16.3±0.2 emu/g. The current study produced a pH­sensitive FA­modified CDDP­loaded MNP that is stable and exhibits magnetic responsiveness, which releases CDDP in a low pH environment.

Star-shaped cyclodextrin-poly(l-lysine) derivative co-delivering docetaxel and MMP-9 siRNA plasmid in cancer therapy.

A new cyclodextrin derivative (CD-PLLD) consisting of a ß-cyclodextrin core and poly(l-lysine) dendron arms was prepared by the click conjugation of per-6-azido-b-cyclodextrin with propargyl focal point poly(l-lysine) dendron of third generation, and then used for docetaxel (DOC) and the best siRNA plasmid targeting MMP-9 (pMR3) co-delivery. Different from commonly used amphiphilic copolymers with cationic character, the as obtained cyclodextrin derivative may be used directly for the combinatorial delivery of nucleic acid and lipophilic anticancer drugs without a complicated micellization process. It was found that CD-PLLD/pMR3 nanocomplex showed a good gene transfection efficiency in vitro, and could mediate the reduce of MMP-9 protein in HNE-1 cells. For co-delivery analysis, the obtained CD-PLLD/DOC/pMR3 complexes could induce a more significant apoptosis than DOC or pMR3 used only, and decreased invasive capacity of HNE-1 cells. Moreover, the star-shaped copolymer exhibited better blood compatibility and lower cytotoxicity compared to PEI-25k in the hemolysis and MTT assays, which should be encouraged in nasopharyngeal cancer therapy.

Staphylococcal enterotoxin B-derived haptens promote sensitization.

T helper 2 (Th2) polarization is a major pathological feature in allergic diseases; its etiology is not fully understood. This study aims to elucidate the adjuvant effect of the microbial product-derived small peptides in the initiation of antigen-specific Th2 polarization. In this study, a clinical survey of patients with chronic rhinosinusitis (CRS) and food allergy (FA) was carried out. The Staphylococcal enterotoxin B (SEB)-derived small peptides (Ssps) were examined in the human stool extracts. The formation of Ssp/antigen adducts was tested in a protein-protein combination assay. The bone marrow-derived dendritic cells (BMDCs) were employed to test the role of Ssp/ovalbumin (OVA) adducts in the dendritic cell (DC) maturation. A mouse model was developed to test the role of Ssp/OVA adducts in the initiation of Th2 polarization in the intestine. The results showed that 54 (18.2%) patients with FA were diagnosed among 296 patients with SEB(+) CRS; only eight (2.9%) FA patients were identified among 272 patients with SEB(-) CRS. Ssps were detected in the stool protein extracts from FA patients with SEB(+) CRS, but not in those with SEB(-) CRS. Ssp/OVA adducts induced DC maturation, speeded up DC migration, activated CD4(+) T cells in the regional lymph nodes and induced skewed Th2 polarization in the local tissue. We conclude that patients with SEB(+) CRS are prone to suffering from FA. SEB can be degraded to Ssps in the gastrointestinal tract. The Ssps can bind macromolecular antigens to form adducts to promote the antigenicity of the antigens and induction of the antigen-specific Th2 polarization and inflammation in the local tissue.

[Development of a hydrophilic anti-fogging agent for nasal endoscopy].

OBJECTIVE: To develop a transparent, non-toxic, non-irritating anti-fogging agent with long-lasting effect for nasal endoscopy. METHODS: The anti-fogging agent was prepared by mixing ethanol, propylene glycol, polyoxyethylene lauryl ether, sodium dodecyl sulfate, polyethylene glycol 400 and deionized water at different proportions based on an orthogonal test design. Twenty-seven test samples of the anti-fogging agents were obtained, which were colorless, transparent, and non-irritating, with a pH value of 7-8. Storz00 nasal endoscopy and its imaging system were used to test the anti-fogging time of the 27 samples, and each agent was tested for 3 times with medical Seoul iodine and 95% ethanol as control. RESULTS: The optimal composition of the anti-fogging agent was 20% ethanol, 10% propylene glycol, 20% polyoxyethylene lauryl ether, 4% sodium dodecyl sulfate, 4% polyethylene glycol, 42% deionized water. The anti-fogging time of this agent reached 15 min, significantly longer than that of medical Seoul iodine (4 min) and 95% ethanol (18 s). CONCLUSION: This anti-fogging agent for nasal endoscopes is colorless and safe and has a long anti-fogging time by forming a homogenous transparent membrane over the endoscopic lens.

Tunable supramolecular hydrogel for in situ encapsulation and sustained release of bioactive lysozyme.

To develop new matrices for the entrapment and sustained release of bioactive lysozyme, a series of supramolecular hydrogels based on α-cyclodextrin (α-CD) and water-soluble poly(ε-caprolactone)-poly(ethylene glycol) block copolymer (PCL-b-PEG) were prepared in the presence of chicken egg lysozyme. Different from commonly used polymeric microspheres and chemically crosslinked hydrogels for lysozyme encapsulation, such hydrogel matrices could be formed under mild conditions without high temperature and the use of chemical emulsifiers or crosslinkers. Their gelation rate, mechanical strength and shear viscosity as well as the release behavior for the encapsulated lysozyme could be tuned easily by the change of α-CD or PCL-b-PEG amount. For the encapsulated lysozyme, its conformation and biological activity could be well maintained when compared to native lysozyme. For the resultant supramolecular hydrogels, they were also confirmed to have a good biocompatibility by MTT assay using mice skin fibroblast (L929).

Modulation of immune tolerance with a Chinese traditional prescription inhibits allergic rhinitis in mice.

BACKGROUND: Allergic diseases substantially affect human health and social economy. The pathogenesis is to be further understood. The effect of current therapeutic remedies on allergic diseases is not satisfactory. AIMS: This study aimed to inhibit allergic rhinitis in a mouse model with a Chinese traditional medical prescription, Bu-Zhong-Yi-Qi-Tang. MATERIAL AND METHODS: A mouse AR model was developed with ovalbumin (OVA) plus adjuvant alum. The AR clinical symptoms and immune pathology in the nasal mucosa were assessed with the AR mouse model. Some mice were treated with Bu-Zhong-Yi-Qi-Tang via gavage-fed. The immune tolerance status in the nasal mucosa was evaluated by counting the numbers of tolerogenic dendritic cells (DC) and regulatory T cells (Treg). RESULTS: After exposure to the specific antigen, OVA, the sensitized mice had AR-like symptoms including nasal itch and sneeze. The frequency of mast cells, levels of IgE/IL-4 in nasal mucosa was markedly higher in sensitized mice than naïve controls; while the levels of integration alphavbeta6 (avb6), the number of tolerogenic DCs and Tregs in nasal mucosa were significantly lower than naïve control mice. The AR-like symptoms and immune pathology and immune tolerance status in the AR nasal mucosa were substantially improved by administration with Bu-Zhong-Yi-Qi-Tang. CONCLUSIONS: The immune tolerance status is impaired in the AR nasal mucosa that can be improved by administering with Bu-Zhong-Yi-Qi-Tang.

[Timing of operation and therapeutic effect of endoscopic optic nerve decompression for traumatic optic neuropathy].

OBJECTIVE: To observe the optimal timing of operation and the therapeutic effect of endoscopic optic nerve decompression for traumatic optic neuropathy (TON). METHODS: The clinical records of 90 consecutive patients with TON (93 eyes) after head and/or maxillofacial trauma from April 1998 to March 2007 were reviewed and analyzed. All patients were either unresponsive or intolerant to medication before they underwent intranasal endoscopic optic nerve decompression. The time interval between the injury and operation ranged from one day to 97 days (median 5.5 days). Among the 93 eyes, there were 71 eyes with no visual acuity before operation and 22 eyes with residue visual acuity, including light perception in 1 eye, hand movement in 5 eyes, counting fingers in 13 eyes, 0.04 in 1 eye, and 0.1 in 2 eyes. Duration of follow-up ranged from 6 days to two years (median 8 days). RESULTS: After decompression, 35 patients (36/93 eyes, 38.7%) showed improvement of visual acuity, 53 patients (55 eyes, 59.1%) remained the same as before operation, while 2 patients (2 eyes, 2.2%) showed decreased visual acuity. Among patients with visual acuity beyond light perception before decompression, 68.2% of them (15/22 eyes) experienced visual improvement, whereas only 22.9% (8/35 eyes, 0.02 in two eyes) among patients who lost visual acuity immediately after injury, and 36.1% (13/36 eyes, 0.02 in five eyes) among those who lost visual acuity gradually after injury. There was a significant difference in visual improvement between group with visual acuity and group with no visual acuity (chi(2) = 11.864, P < 0.01). Among patients with no visual acuity, 41.2% of those (7/17 eyes) who underwent operation within 3 days of injury, experienced improvement in visual acuity, compared with 25.9% (14/54 eyes) for those who underwent the operation more than 3 days after injury. It was indicated that no significant difference in visual improvement between these two groups (chi(2) = 1.46, P > 0.05). When comparing different sites of fracture, the effect of surgery was the most desirable (55.6%, 10/18 eyes improved) if the fracture occurred simultaneously in both exterior and interior walls of optic canal, followed by the interior wall fracture (45.7%, 21/46 eyes). The operation was less effective if there was no fraction (20%, 4/20 eyes) or if the fracture occurred in exterior wall alone (11.1%, 1/9 eyes). CONCLUSIONS: Endoscopic optic nerve decompression is a minimally invasive procedure with no adverse cosmetic effects. Early operation is recommended for saving vision, even though visual acuity is lost immediately after injury. However, the satisfactory clinical effects of endoscopic optic nerve decompression require further study.

[Retrospective analysis of the effect of intranasal endoscopic resection of olfactory neuroblastoma].

OBJECTIVE: To investigate the effectiveness of intranasal endoscopic resection of olfactory neuroblastoma and the significance of assistant radiotherapy. METHODS: Six patients (4 men and 2 women) ranging in age from 9-68 years (median age, 48 years) with olfactory neuroblastoma treated endoscopically at the Third Affiliated Hospital of SUN Yat-sen University between August 2001 and September 2005 were retrospectively analyzed. The Kadish clinical stage was determined for all six cases, two cases with stage B, and 4 with stage C. The duration of endoscopic follow-up ranged from fourteen months to sixty three months. RESULTS: The tumor in five patients was completely excised including one case underwent craniotomy firstly and one case underwent neck dissection and the another case had her tumor subtotally resected. Five cases underwent assistant radiotherapy after operation without any evidence of recurrence so far, while the other one (Kadish stage B, tumor totally resection), who had no radiotherapy after first procedure, died from local recurrence and cerebellar metastasis 31 months after operation. The medium survival duration was 44.5 months. CONCLUSIONS: With simple approach, excellent visualization, a less operative invasiveness and no scar on face, the endoscopic approach appears to be the method of choice for the treatment of olfactory neuroblastoma. But it was necessary for good prognosis to combine with radiotherapy after operation.

[Morphological study of adenoid by endoscopy and its clinical significance].

OBJECTIVE: To observe the natural process of adenoid growth and degeneration as the age grows, to investigate the related clinical significance and pathologic characteristics of hypertrophied adenoid. METHODS: Totally 2650 (age 2 to 87) cases with nasal obstruction or/and other symptoms were included in the patients group, and 810 (age 3 to 85) subjects without symptoms were included as the control group. Morphological characteristics examined with nasal endoscope. Biopsy was performed for 39 cases. The adenoid was calcified as 4 degrees according to the size. RESULTS: In the patient group, age 2 to 9, degree III and degree II adenoid were 81.1% (198/244) and 18.9% (46/244) respectively. And adenoid of children whose age 2 to 5 was 100.0% in degree III; In above 10 years old group, the adenoid was mostly degree II. In age 60 to 69 group, degree 0 was (66.5%), and in age 81 or above, degree 0 reaches 100%. And 19 years old was the youngest age at which adenoid of degree 0 started to be found and 21 was the oldest age at which there is no adenoid of degree III. In the control group, compared with the patient group, no statistical significant difference found in all other groups except in age 2 to 9 (degree III 57.9%, 22/38, degree II 42.1%, 16/38). Shapes of adenoids at degree II varied while degree I were almost like peeled orange. Pathologically, among children there are abundant of adenoidal lymph tissue, while in adults the lymph tissue getting less as age grows but with evident inflammation reaction. Among patients, the incidence of sinusitis and snoring was higher in degree III group compared with others, 47.4% and 18.7% respectively, and the differences is statistically significant (chi2 = 51.28, P < 0.01; chi2 = 40.26, P < 0.01). CONCLUSIONS: Adenoid volume of children (age < 10) is the biggest, especially of children under 5 years old.

[Transnasal endoscopic repair of nasal-skull base defect: 69 cases].

OBJECTIVE: To summarize the experience of transnasal endoscopic repair of nasal-skull base defect. METHODS: The clinical data of 69 patients with transnasal endoscopic repair of skull base defect from January 1998 to June 2006 were analyzed retrospectively. The causes, sites of defect and the effect and key technique of transnasal endoscopic repair were investigated. RESULTS: All patients were followed-up for 6 months to 11 years. The causes were mainly traumatic fracture, occupying lesions and iatrogenic trauma. The sites were mainly in ethmoid sinus and sphenoid sinus. The success rate of one attempt was 91.3% (63/69), the ultimate success rate was 95.7% (66/69). CONCLUSIONS: Transnasal endoscopic repair of skull base defect may be the best approach. Debriding and raw edge of defect area and "inlay-plug" method are of key technique. The liber muscle and fascia are ideal repair material.

[Glucocorticoid receptor subunit expression in adenoid tissue of children with obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: The expression of glucocorticoid receptor (GR) expression in obstructive sleep apnea hypopnea syndrome (OSAHS) in children is currently unknown. The aim of this study was to determine the GR-alpha and GR-beta status in the adenoidal tissues in children with OSAHS. METHODS: Thirty-four pediatric patients (aged 3-14 years, median 7.8 years) had sleep study with polysomnography before adenoidectomy. According to the criteria of apnea hypopnea index (AHI) > or = 5 /h or/and apnea index (AI) > or = 1/h, they were divided into OSAHS and non-OSAHS sub groups. The study was based on fluorescent quantitative PCR (FQ-RT-PCR) for the mRNA expression of GR-alpha and GR-beta in the adenoidal tissues in children. RESULTS: GR isoforms mRNA encoding for expression of both GR-alpha and GR-beta were detected in the adenoids of all children. GR-alpha mRNA level [(9.40 +/- 3.06) x 10(5) cDNA copies/microg total RNA] in the adenoidal tissues in OSAHS was lower than those in the non-OSAHS [(1.60 +/- 0.26) x 10(6) cDNA copies/microg total RNA] (F = 40.285, P < 0.001), whereas no differences found for GR-beta [(1.57 +/- 0.35) x 10(4) cDNA copies/microg total RNA, (1.52 +/- 0.18) x 10(4) cDNA copies/microg total RNA]. GR-alpha/GR-beta ratio was 62.3 +/- 20. 3 in OSAHS and 107.4 +/- 24.4 in non-OSAHS. AHI or AI was not related to the mRNA levels of GR-alpha and GR-beta in OSAHS or non-OSAHS. CONCLUSIONS: GR-alpha and GR-beta were detectable in the adenoidal tissues in children. These data indicated that the relationship between the expressions of GR and the clinical significance in OSAHS need further and profound investigation.

[Effect of specific immunoglobulin Y in the treatment of acute and chronic pharyngitis].

OBJECTIVE: To evaluate the effect and safety of the specific immunoglobulin Y (IgY) for treatment of acute and chronic pharyngitis. METHODS: Double-blind, randomized, placebo-controlled trial was conducted on 50 adults with acute pharyngitis. Experimental group received a 6 times-daily total 30 doses of IgY stomat-spray which contained specific immunoglobulin Y (titer = 512) prepared from the egg yolk of hens immunized with a variety of bacteria. Another open label trial included 50 patients, whose ages ranged from 21-69 years, including 25 cases of acute pharyngitis and 25 cases of chronic pharyngitis were also treated using IgY stomat-spray. The therapeutic effect were objectively evaluated 7 days later by the decreased scores based on both the symptoms and physical signs. If the symptom did not improve or became severe three days later, these patients with acute pharyngitis was inefficiency and antibiotic medicine would be added to them. RESULTS: In Double-blind trial, 8 cases (32%) received IgY had apparent effect with the decreased scores 5 or more than 5, 13 cases (52%) had effective with the decreased scores 3-4, and other 4 cases (16%) had inefficacy with the decreased scores only 2 or no more than 2. While in placebo-controlled group, only 2 (8%) cases had apparent effect, 5 (20%) cases showed effective and 18 (72%) cases had non-effect. The difference between the two groups was significant (chi 2 = 16.06, P < 0.01). In open label trial, 19 cases (38%) showed apparent effect, in which 14 cases were acute pharygitis. 23 cases (46%) had effective, in which 10 cases were acute pharyngitis. The left 8 cases (16%) had ineffective, in which one case was acute pharyngitis. There was significantly difference (chi 2 = 8.90, P < 0.05) between acute pharyngitis and chronic pharyngitis. An average of three months followup showed that there were no side effect or toxic effect and no allergic reaction. CONCLUSION: The IgY stomat-spray is a safe and effective agent in treating acute and chronic pharyngitis, especially for acute pharyngitis.

[Relationship of matrilysin with invasion, metastasis, and prognosis in laryngeal cancer].

BACKGROUND & OBJECTIVE: Matrilysin (matrix metalloproteinases-7, MMP-7) might play an important role in invasion and metastasis of tumor and is thought to be related to prognosis of many kinds of carcinomas. This study was designed to investigate the significance of MMP-7 expression in the invasion and metastasis of laryngeal cancer and the relationship of MMP-7 expression with the prognosis of laryngeal cancer. METHODS: The expression of MMP-7 in 70 samples with paraffin-embedded laryngeal cancer and corresponding para-tumor normal mucosas were using immunohistochemical technique. The mRNA expression of MMP-7 were examined by reverse transcription-polymerase chain reaction (RT-PCR) in 24 frozen specimens of laryngeal cancer and corresponding para-tumor normal mucosas. RESULTS: (1) The immunohistochemical staining of MMP-7 was observed in 77.1% (54/70) of cancer tissues, which was significantly higher than that (5.7%, 4/70) of para-tumor normal mucosas (P< 0.01). The mRNA expression of MMP-7 was detected in 75.0% (18/24) of cancer tissues, which was also significantly higher than that (4.2%, 1/24) of para-tumor normal mucosas (P< 0.01). (2) The expression of MMP-7 in protein level was hot only higher in T3-T4 group than that in T1-T2 group (P< 0.01) but also higher in nodal metastases group than that in group without nodal metastases (P< 0.01). (3) The mRNA expression of MMP-7 was significantly higher in T3-T4 group than that in T1-T2 group (2.908 +/- 0.891 vs.1.662 +/- 0.508, P< 0.01) and was significantly higher in nodal metastases group than that in group without nodal metastases (2.958 +/- 0.827 vs. 1.887 +/- 0.769,P< 0.01). (4) Kaplan-Meier survival curve showed that the prognosis of group with MMP-7 protein high expression was poorer than that in group of MMP-7 protein weak or negative expression (log-rank=4.7559, 8.9513; P=0.0292, 0.0028). No significant difference was found between group with MMP-7 protein weak expression and group with that negative expression (log-rank=0.0314, P= 0.8593). CONCLUSION: MMP-7 possesses close relationship with the invasion, metastasis, and prognosis of laryngeal cancer, and it may be served as a marker in estimating the invasive and metastatic potency and prognosis of laryngeal cancer.

[Detection of chromosomal aberrations in nasopharyngeal carcinoma by rapid primed in situ labeling].

BACKGROUND & OBJECTIVE: The chromosomal aberration is common in nasopharyngeal carcinoma (NPC). The currently-used methods for detecting chromosomal abnormalities are complicated and of limited clinical value. The purpose of the present study was to explore the feasibility of detecting chromosomal abnormalities in NPC by rapid primed in situ labeling (RPRINS). METHODS: Using RPRINS technique with specific oligonucleotide primers of chromosome 3 and 7, the abnormalities of chromosome 3 and 7 in the frozen section tissues of 15 cases of NPC and 5 cases of normal nasopharyngeal mucosa (NNM) were detected. Loss of chromosome was defined when the percentage of cells with labeling signals /=65%, and the increase in chromosomal copy number was defined when the percentage of cells with labeling signals >/=3 was >/=6.5%. RESULTS: In 15 cases of NPC tissues, the chromosome 3 had a labeling rate of 88.6% and increasing copy numbers in ten cases (66.7%), and the chromosome 7 had a labeling rate of 87.4% and loss of chromosome in five cases (33.3%). Four cases coexisted with increasing chromosome 3 copy numbers and loss of chromosome 7. In contrast, the labeling rates of chromosome 3 and 7 in NNM were 92.0% and 91.8%, respectively, and the percentage of diploid cells were 43.2% and 43.6%, respectively, with absence of triploid. There was a significant difference in the percentage of diploid cells between NPC and NNM (P< 0.05). CONCLUSION: The technique of rapid PRINS could be used to detect chromosomes in frozen section tissues, and the chromosomal abnormalities would be helpful in diagnosis of NPC.

[Treatment of endonasal-encephalomeningocele by transnasal endoscopic surgery].

OBJECTIVE: To inquire into the feasibility of transnasal endoscopic surgery for endonasal encephalomeningocele, and to put forward the basic management guidelines. METHOD: Five cases of endonasal encephalomeningocele were managed by transnasal endoscopic surgery. Different surgical techniques were applied according to the size and shape of skull base defect. At the same time, one-stage repair of skull base defect was achieved by applying muscle, fascia, cartilage and bone. RESULTS: All 5 patients were successfully treated without complication and relapse during 1-5 year follow-up. CONCLUSION: Endonasal type encephalomeningocele can be managed by transnasal endoscopic surgery. If the skull base defect is less than 0.5 cm, it can be repaired by packing muscle and fascia. If the defect is larger than 0.5 cm, the cartilage or bone should be used to repair the skull base.