Healing of the Torn Anterior Horn of Rabbit Medial Meniscus to Bone after Transtibial Pull-Out Repair and Autologous Platelet-Rich Plasma Gel Injection.

OBJECTIVES: The transtibial pull-out repair (TP) is a relatively new method for treating meniscal root tear; however, the clinical evaluation of its healing effect remains controversial. Due to ethical constraints and limitations of imaging techniques in humans, here we dynamically observe the healing effects of TP and TP with platelet-rich plasma gel (PRG) at the histological level using an animal model. METHODS: Platelet-rich plasma (PRP) and PRG of rabbits were prepared. Platelet-derived growth factor (PDGF) and transforming growth factor-ß1 (TGF-ß1) levels in PRP and PRG were determined using an enzyme-linked immunosorbent assay. A rabbit model of anterior horn tear of the medial meniscus and TP surgery were created. PRG was injected between the anterior horn of the medial meniscus and the tibial tunnel. Rabbits were divided into three groups: the anterior horn tear group (Tear group), the anterior horn tear + TP group (TP group), and the anterior horn tear + TP + PRG group (TP + PRG group). The healing effect was observed dynamically using histopathological studies and biomechanical experiments. RESULTS: The platelet content in PRP significantly increased to approximately 4.57 times that of whole blood. PDGF and TGF-ß1 concentrations in PRG increased to 2.46 and 4.15 times those in PRP, respectively. Hematoxylin and eosin (H&E) and Masson staining showed that the number of inflammatory cells in healing tissue decreased and the collagen fibers significantly increased in TP and TP + PRG groups at 4, 8, and 12 weeks postoperatively compared to those in Tear group. Neatly arranged, interlaced, and dense collagen fibers were found between the anterior horn and bone at 12 weeks. H&E and toluidine blue staining showed that the injury to the femoral condyle cartilage was alleviated. The healing performance in TP + PRG group was better and faster than that in TP group. The maximum tensile fracture strength of the meniscus progressively increased at 8 and 12 weeks postoperatively. CONCLUSIONS: Anterior horn injury of the medial meniscus in rabbits can be repaired using the TP technique, and the addition of autologous PRG to the bone tunnel promotes early healing of the meniscus and bone postoperatively. Meanwhile, both treatments can reduce the secondary damage to the cartilage due to osteoarthritis.

SHR-A1403, a novel c-Met antibody-drug conjugate, exerts encouraging anti-tumor activity in c-Met-overexpressing models.

Emerging evidence demonstrates that a c-Met antibody-drug conjugate (ADC) has superior efficacy and safety profiles compared with those of currently available small molecules or antibody inhibitors for the treatment of c-Met-overexpressing cancers. Here we described both the in vitro and in vivo efficacies of SHR-A1403, a novel c-Met ADC composed of a humanized IgG2 monoclonal antibody against c-Met conjugated to a novel cytotoxic microtubule inhibitor. SHR-A1403 showed high affinity to c-Met proteins derived from human or monkey and potent inhibitory effects in cancer cell lines with high c-Met protein expression. In mice bearing tumors derived from cancer cell lines or patient HCC tissues with confirmed c-Met overexpression, SHR-A1403 showed excellent anti-tumor efficacy. Antibody binding with c-Met contributed to SHR-A1403 endocytosis; the subsequent translocation to lysosomes and cytotoxicity of the released toxin are speculated to be predominant mechanisms underlying the anti-tumor activity of SHR-A1403. In conclusion, SHR-A1403 showed significant anti-tumor activity in cancer cell lines, xenograft mouse models and an HCC PDX model, which all have high c-Met levels. These data provide references for SHR-A1403 as a potential therapy for the treatment of cancers with c-Met overexpression.

Discovery and Modification of in Vivo Active Nrf2 Activators with 1,2,4-Oxadiazole Core: Hits Identification and Structure-Activity Relationship Study.

Induction of phase II antioxidant enzymes by activation of Nrf2/ARE pathway has been recognized as a promising strategy for the regulation of oxidative stress-related diseases. Herein we report our effort on the discovery and optimization of Nrf2 activators with 1,2,4-oxadiazole core. Screening of an in-house collection containing 7500 compounds by ARE-luciferase reporter assay revealed a moderate Nrf2 activator, 1. Aimed at obtaining more derivatives efficiently, molecular similarity search by the combination of 2D fingerprint-based and 3D shape-based search was applied to virtually screening the Chemdiv collection. Three derivatives with the same core were identified to have better inductivity of Nrf2 than 1. The best hit 4 was selected as starting point for structurally optimization, leading to a much more potent derivative 32. It in vitro upregulated gene and protein level of Nrf2 as well as its downstream markers such as NQO1, GCLM, and HO-1. It remarkably suppressed inflammation in the in vivo LPS-challenged mouse model. Our results provide a new chemotype as Nrf2-ARE activators which deserve further optimization with the aim to obtain active anti-inflammatory agents through Nrf2-ARE pathway.

SHR3824, a novel selective inhibitor of renal sodium glucose cotransporter 2, exhibits antidiabetic efficacy in rodent models.

AIM: The sodium glucose cotransporter 2 (SGLT2) plays an important role in renal glucose reabsorption, thus serves as a new target for the treatment of diabetes. The purpose of this study was to evaluate SHR3824 as a novel selective SGLT2 inhibitor and to characterize its in vivo effects on glucose homeostasis. The effects of chronic administration of SHR3824 on peripheral insulin sensitivity and pancreatic ß-cell function were also investigated. METHODS: The in vitro potency and selectivity of SHR3824 were assessed in HEK293 cells transfected with human SGLT2 or SGLT1. Acute and multi-dose studies were performed on ICR mice, GK rats and db/db mice to assess the ability of SHR3824 to enhance urinary glucose excretion and improve blood glucose levels. 2-Deoxyglucose uptake and insulin immunohistochemical staining were performed in the soleus muscle and pancreas, respectively, of db/db mice. A selective SGLT2 inhibitor BMS512148 (dapagliflozin) was taken as positive control. RESULTS: SHR3824 potently inhibited human SGLT2 in vitro, but exerted much weak inhibition on human SGLT1 (the IC50 values of SHR3824 against human SGLT2 and SGLT1 were 2.38 and 4324 nmol/L, respectively). Acute oral administration of SHR3824 (0.3, 1.0, 3.0 mg/kg) dose-dependently improved glucose tolerance in ICR mice, and reduced hyperglycemia by increasing urinary glucose excretion in GK rats and db/db mice. Chronic oral administration of SHR3824 (0.3, 1.0, 3.0 mg·kg(-1)·d(-1)) dose-dependently reduced blood glucose and HbA1c levels in GK rats and db/db mice, and significantly increased insulin-stimulated glucose uptake in the soleus muscles and enhanced insulin staining in the islet cells of db/db mice. CONCLUSION: SHR3824 is a potent and selective SGLT2 inhibitor and exhibits antidiabetic efficacy in several rodent models, suggesting its potential as a new therapeutic agent for the treatment of type 2 diabetes.

Clinical outcomes of endoscopic and open resection of recurrent sinonasal inverted papilloma.

PURPOSE: The aim of this study was to investigate the effectiveness of endoscopic surgery for recurrent sinonasal inverted papilloma and evaluate the recurrence rates of endoscopic and open resection at a single institution. METHODS: This retrospective study was performed on 21 patients with histopathologically confirmed recurrent sinonasal inverted papilloma who underwent an operation in our department from January 1990 to January 2005. The 21 recurrent cases were categorized according to the Krouse staging system into 1 case of stage 1 (5%), 18 cases of stage 2 (85%), 2 cases of stage 3 (10%), and 0 cases of stage 4 (0%). There were 7 patients (33%) who underwent endoscopic resection and 14 patients (67%) who underwent open resection. RESULTS: Of the 21 recurrent patients who underwent resection, 4 patients (19%) were found to have recurrence. The mean time to recurrence was 28 months. The recurrence rates of the patients with stage 1, stage 2, and stage 3 were 0%, 17%, and 50%, respectively (P < 0.05). Recurrence was observed in 1 patient (14%) in the endoscopic group and 3 patients (21%) in the open group (P > 0.05). There was statistical difference in recurrence rates between stage 1 and stage 2 in the endoscopic group (P < 0.05) and between stage 2 and stage 3 in the open group (P < 0.05). There was no significant difference in recurrence rates between the endoscopic and the open group in stage 2 (P > 0.05). CONCLUSIONS: Endoscopic and open approaches are both available to achieve radical excision of the recurrent sinonasal inverted papilloma, with similar rates of recurrence. The risk for recurrence is likely related to the Krouse stage of the tumor, with more aggressive tumors having a higher propensity for recurrence. Endoscopic surgery is an effective treatment of recurrent sinonasal inverted papilloma.

[Effect of vasopressin on aquaporin 7 expression in rat inner ear].

OBJECTIVE: To study the effect of vasopressin on aquaporin 7 (AQP7) expression in rat inner ear and reveal the possible role of aquaporins in the formation of endolymphatic hydrops induced by vasopressin. METHODS: Wistar rats were intraperitoneally injected with 50 microg/kg arginine vasopressin once a day for one week. Differentially expressed genes of aquaporins induced by vasopressin injection in rat inner ear were filtered by cDNA microarray. The changes of mRNA expression level of AQP7 in inner ear of rats treated with vasopressin injection were measured by RT-PCR. RESULTS: Differentially expressed gene AQP7 of aquaporins induced by vasopressin injection was screened out in rat inner ear. The expression level of AQP7 mRNA in inner ear of rats treated with vasopressin injection was significantly lower. CONCLUSION: Vasopressin may down-regulate the expression of AQP7 mRNA in the endolymphatic sac and induce a decreased absorption of endolymph, which decreases the water permeability in the potassium ions recycle pathway in the organ of Corti and disturbs the circulation of endolymph, resulting in endolymphatic hydrops.

[Clinical analysis of normal subjects and dizzy patients with postural sway test].

OBJECTIVE: To study the characteristics of the normal subjects with postural sway test and discuss the value of postural sway test in the diagnosis of dizzy patients. METHODS: Totally 112 normal subjects, 72 patients with peripheral vertigo, and 30 patients with central vertigo were examined using a stabilometer (EAB-100, Anima Co., Japan). Items include patterns, length of locus, envelope area, deflection average center displacement, and romberg rate were recorded and compared. RESULTS: The postural sway was classified into the following five patterns: centripetal, forward and backward, right and left, diffuse, and multicentric. Centripetal pattern was the most prominent figure pattern in the normal subjects, while diffuse pattern was the most prominent figure pattern in the patients with central vertigo. When the normal subjects closed their eyes, age was linearly correlated with the length of locus, envelope area, rectangel area, and unit area path length (P < 0.05). Sex was not correlated to all of these parameters. The length of locus and envelope area in the patient groups were significantly increased when compared with the normal subjects (P < 0.0167). Romberg rate was not significantly different between the patients and the normal subjects. CONCLUSIONS: The pattern of postural sway is useful for the diagnosis of vertigo. Age is an important factor that affects the balance function. Sex seems has no influence. Visual input plays an important role in maintaining postural balance. Stabilometer is a useful tool for evaluating balance function, and postural sway test may be a valuable assistant examination.

Effects of vasopressin on gene expression in rat inner ear.

Vasopressin regulates water excretion from the kidney by increasing water permeability of the collecting duct as a hormone secreted from the posterior pituitary. A clinical study reported that plasma levels of arginine vasopressin were significantly higher in patients suffering from Meniere's disease. It was histologically confirmed that chronic administration of vasopressin induced endolymphatic hydrops in guinea pigs. However, the mechanism of endolymphatic hydrops induced by vasopressin is still unclear. We use cDNA microarray to study the effects of vasopressin on gene expression profiles in rat inner ear to elucidate the possible mechanism of the induced hydrolabyrinth. Wistar rats were intraperitoneally injected with 50 microg/kg arginine vasopressin once a day for one week. Hydrolabyrinth in rat inner ear induced by administration of vasopressin was detected by HE stain. The bullae were dissected out for total RNA extraction. cDNAs were synthesized by reverse transcription and labeled with Cyanine3 (Cy3) or Cyanine5 (Cy5). The BiostarR-40s cDNA microarray was hybridized with the above cDNAs and the changes of mRNA expression intensity were showed by data analysis. Furthermore, the changes of aquaporins expression level were measured by reverse transcription polymerase chain (RT-PCR). Endolymphatic hydrops were present in rats intraperitoneally injected with vasopressin. 226 known differentially expressed genes were screened out in rat inner ear induced by vasopressin injection. Of the 226 genes, 18 transcripts were increased by 5-fold or more, and 7 transcripts were decreased to 0.2-fold or less. Ten differentially expressed genes were identified that associate with cell signal transduction, 14 differentially expressed genes were identified that relate to ion transport, 7 differentially expressed genes were involved in vesicle-mediated transport, and 2 differentially expressed genes were aquaporin 2 (AQP2) and aquaporin 7 (AQP7). The expression level of AQP2 was significantly higher and AQP7 was significantly lower. These results suggest that there are obvious differences in gene expression profiles in inner ear between vasopressin injected rats and normal control rats. Vasopressin may disturb fluid homeostasis in inner ear by way of signal transduction, ion transport, vesicle-mediated transport and aquaporins. It is likely that up-regulated expression of AQP2 mRNA and down-regulated expression of AQP7 mRNA in the rat inner ear caused by vasopressin induce an increased production and a decreased absorption of endolymph, resulting in endolymphatic hydrops.

[Isolation and identification of arboviruses in Hebei Province].

BACKGROUND: To study the arboviruses carried by mosquitoes collected in Hebei Province. METHODS: Samples were collected from mosquito active sites and stored in liquid nitrogen till use. Pools of 20 to 30 mosquitoes were ground after sterilization, centrifugal supernant was inoculated onto C6/36 cell, cytopathic effect was observed for three sequential passages. Positive isolates were identified by IFA and RT-PCR. RESULTS: Totally 1310 mosquitoes were collected from two villages of She county, Hebei province. They were divided into 46 pools and ground respectively. Thirteen positive isolates were obtained. Two isolates reacted with alphaviral antibodies and were amplified by alphaviral primers, nucleotide sequence showed the highest homology (98%) to Getah virus (AY702913.1), so the two isolates were identified as Getah virus. CONCLUSION: Getah virus was isolated from mosquitoes in Hebei Province. This is the first report of isolating Getah virus from inland of China.

Post-inflammatory glomerular remodeling is influenced by transformed mesangial cells.

To identify the role of transformed mesangial cells (MC) during glomerular remodeling, anti-thymocyte-1 (Thy1) nephritis; modified Thy1 nephritis (injections of anti-Thy1 antibody four times, weekly); and Thy1 nephritis treated with signal transduction inhibitor 571 (Thy1 + STI); were analyzed. At week 1 the index of MC proliferation in modified Thy1 nephritis and in mesangiolysis in Thy1 + STI nephritis was highest among the three models. From week 4, the index of alpha-smooth muscle actin (alpha-SMA) was significantly higher in modified Thy1 nephritis than the other two models. Production of the mesangial matrix including type IV collagen was increased in modified Thy1 but inhibited in Thy1 + STI nephritis. In contrast to modified Thy1 nephritis, the capillary numbers in glomeruli recovered to normal at week 4 in Thy1, and at week 8 in Thy1 + STI nephritis. At week 12, both the adhesive and sclerotic index was significantly higher in modified Thy1 than in the other two models. Data suggest that a moderate amount of mesangial matrix results in a complete repair of capillary loops. Overproduction of the mesangial matrix retards capillary remodeling and finally induces glomerulosclerosis. Insufficient mesangial matrix delays the repair of capillary loops. In conclusion, transformed MC may influence glomerular remodeling by changing the amount of mesangial matrix.

[Effect of nitric oxide in vestibular compensation].

OBJECTIVE: To study the effect of nitric oxide (NO) in vestibular compensation after unilateral vestibular deafferentation. METHODS: Eighteen animals were divided into two groups, 6 of group a as control, 12 of group b received gentamicin intratympanic injection in the left ear. Half of the animals were killed respectively after 5 days and 10 days. Vestibular endorgan and brainstem tissue sections were subjected to NADPH-d reactive test of NOS for histochemical examination. RESULTS: In group a, NOS-like reactivity in both sides of vestibular endorgan and nucli. In group b during 5 days, NOS-like reactivity in right side of vestibular endorgan and nucli, those of the left side were negative. During 10 days, NOS-like reactivity only in the right side of vestibular endorgan. CONCLUSIONS: Changes of NOS expression in the contralateral vestibular nucli might have played a role in vestibular compensation.

[Pharyagoesophageal sphinoter myotomy for voice rehabilitation after total laryngectomy].

OBJECTIVE: In those patients received total laryngectomy, Blom-Singer prosthesis speech appears to be the more effective form of rehabilitation. To reduce the effect of pharyngoesophageal sphincter on the Blom-Singer prosthesis voice restoration following total laryngectomy, the pharyagoesphageal sphincter myotomy was designed and carried out. METHODS: After total laryngectomy, the cricopharyngeus muscle stricture or spasm appears to inhibit the air flowing and prevent the speech. The pharyngoesophageal sphincter is now defined as the area including the cricopharyngeus with a portion of inferior constrictor, as well as some of the upper cervical esophagus. The pharyngoesophageal sphincter were incised intra- and post- operation of total laryngectomy, the dissection was approximately 5 cm in vertical length and 1 cm in width. RESULTS: Among the 33 patients who received pharyngoesophageal sphincter myotomy following total laryngectomy, voice restoration was achieved in 32 cases, the success rate was 97%. 25 patients had survived for 3 years, 18 patients survived for 5 years. CONCLUSION: Pharyagoesophageal sphincter myotomy can raise the success rate of rehabilitation of the voice by Blom-Singer prosthesis.

[The use of muscle autograft denatured by microwave for repair of gaps in removal of facial neuromas].

OBJECTIVE: To study the clinical value of muscle autograft denatured by microwave for repair of gaps in removal of facial neuromas. METHODS: Two cases of patients with facial nerve Schwann cell neuromas were reported. The operations for removal of facial neuromas were completed, and the gaps of the nerves were repaired with muscle autograft denatured by microwave of 250 W for 120 sec. RESULTS: The patients were followed up for two years, and the recovery of facial function on the affected sides were satisfactory. CONCLUSION: Muscle autograft denatured by microwave technique is convenient, highly efficient for repairing facial nerve gap after removal of facial neuroma.

[Surgical management of metastatic disease in the conjunctive area between neck and thorax].

OBJECTIVE: To study the surgical management of metastatic disease in the conjunctive area between the neck and thorax and its efficacy. METHODS: Fourteen cases with metastatic node disease in the area between neck and thorax were collected and analysed. Eleven tumors were from the thyroid cancer, and the other three were from the hypopharyngeal cancer, esophagual cancer and malignant pheochromocytoma, respectively. The clavicle was displaced or resected, and the upper half of the manubrium might also be resected when necessary. The recurrent laryngeal nerve and phrenic nerve were exposed and protected. The metastatic disease was completely removed with the internal jugular and/or the brachiocephalic vein resected or spared, depending on the disease condition. RESULTS: In 10 cases with metastases from the thyroid, no local recurrence was found within the follow-up period from 2 to 5 years. In contrast, no patient with metastatic disease from hypopharyngeal or esophageal cancer survived more than 11 months. No serious complications were found in this group. CONCLUSIONS: The surgical treatment of node metastases in the conjunctive area between neck and thorax from the well-developed thyroid cancer has promising effect and is comparatively safe.

[Endolymphatic hydrops of both ears of guinea pig produced by aldosterone].

OBJECTIVE: To develop endolymphatic hydrops in both ears of guinea pigs by aldosterone. METHODS: Thirty animals were divided into 3 groups with 10 in each group. Group A received left ears operation following Kimura', with right ears non-operated as control; group B received introperitonal injection of aldosterone (0.1 mg.0.1 kg-1.d-1) for continuous 5 days; group C received introperitonal injection of 0.5 ml 0.9% NS. Five out of group A,B and C were killed 1 month and 2 months later respectively for analysis of blood K+, Ca2+, Na+ and pathological examination of inner ear, heart, lung, brain, liver and kidney. RESULTS: In group A' moderate to severe endolymphatic hydrops developed in the operated ears during 1 and 2 months. In group B, mild endolymphatic hydrops developed in the both ears during 1 month, and within 2 months it was moderate and severe. No endolymphatic hydrops developed in group C. In contrast to group C, No significant changes could be found in blood analysis of group A, but the concentrations of K+ and Ca2+ were declined, and that of Na+ were increased in group B. The heart, lung, brain, liver, and kidney of all animals were normal. CONCLUSIONS: Aldosterone may induce endolymphatic hydrops in both ears.

[The use of muscle autograft denatured by microwave for repair of gap in removal of facial neuroma].

OBJECTIVE: To study the clinical value of muscle autograft denatured by microwave for repair of gap in removal of facial neuroma. METHODS: A case of patient with a larger facial nerve Schwann cell neuroma was reported. Based on the good results of our experimental research on rats recently, we completed the operation by transmastoid approach for removal of facial neuroma, and the gap of the nerve was repaired with muscle autograft denatured by microwave. RESULTS: The patient was followed up for one year, and her facial function recovery on the affected side was satisfactory. CONCLUSION: Muscle autograft denatured by microwave technique is convenient, highly efficient for repairing facial nerve gap after removal of facial neuroma.