Objective To investigate the regrlative role of the cervical sympathetic ganglia on the cochlear blood flow and auditory function in guinea pig.Methods The horseradish peroxidase(HRP) was given to spiral modiolar artery locally for retrograde tracing in guinea pig. Immunohistochemical double-labeled technique was used in this study. The cochlear blood flow and auditory brainstem response(ABR) was measured a week after anilateral superior cervical sympathectony. The animal model of superior cervical sympathectomy following noise exposure in guinea pigs was estableshed to observe the auditory threshold shift.Results Retrogradely labeled neurons with HRP were found in the ipsilateral superior cervical ganglion(SCG). Most of HRP-labeled neurons in SCG were tyrosion hydroxylase(TH) positive. The blood flow of the capillaries of the stria vascularis on the experimental side a week ago were more aplenty than that of the opposite side, but the ABR threshold did not changed before and after ablation of the superior cervical ganglion.A protective role against noise injury was observed after surgical ablation of superior cervical ganglion.Conclusion The superior cervical sympathectomy can influence the cochlear blood flow and auditory function in guinea pig.

BACKGROUND: Spinal cord can regenerate after injury in certain microenvironment. Olfactory ensheathing cells (OECs)have the characteristics of astrocytes and Schwann cells and can accelerate the spinal cord axonal regeneration.OBJECTIVE: To make injured thoracic cord rat models and observe the effect of OECs on injured spinal cord axonal regeneration.DESIGN: Observational experiment.SETTING: Second Affiliated Hospital of Sun Yat-Sen University.MATERIALS: The experiment was performed at the Second Affiliated Hospital of Sun Yat-Sen University from January 2001 to November 2002.Totally 20 adult SD male rats with the body mass of (380±20) g were provided by Experimental Animal Center of Sun Yat-Sen University (number of institution license SYXK2004-0020). There were DMEM culture solution with low glucose (L-DMEM, GibcoBRL), fetal calf serum (FCS) (Hyclone), myelin basic protein (MBP) (Sigma) and nerve growth factor receptor antibody (Sigma). They were divided into cell transplantation group and control group by the method of random digits table with 10 in each group.METHODS: The adult SD rats were anaesthetized and decapitated to obtain the whole olfactory bulb and then isolate olfactory nerve with a sterile operation. Thoracic cord injury models were established by modified Allen method. 10μL OECs suspension (2.5×1010 L-1) was injected into injured spinal cord of the cell transplantation group, whereas DMEM/F12 (1:1) culture solution of the same dose was injected in the control group. The influence of OECs on spinal cord axonal regeneration was observed by histological and immunohistochemical method 6 weeks after transplantation.MAIN OUTCOME MEASURES: ①OECs were identified by nerve growth factor receptor antibody staining. ②Repair of myelin sheath was observed by MBP staining. ③Nerve axonal regeneration was observed by argentaffin staining. RESULTS: Two rats in the cell transplantation group and 3 rats in the control group died, so totally 15 rats were involved in the result analysis. ①In the cell transplantation group,injured spinal meninges were integrated,but spinal cord became thin as compared with the normal spinal cord. By hematoxylin-eosin (HE) staining, multipolar cells appeared in injured region and the cells were fused excessively with myeloid tissues. It proved that survival OECs were integrated well within the host. New axons were clustered in bundles and infiltrated by small round lymphocytes. By argentaffin staining, regenerated axons grew in tissues of injured region, which mostly accompanied with fascicular-arranged multipolar cells. In the control group, spinal cord became thin markedly and spinal meninges were integrated. No new axon appeared in the injured spinal cord by HE staining. No regenerated axon appeared by argentaffin staining, either. ②In the cell transplantation group, most multipolar cells were clustered in bundles. A mass of positive granules of nerve growth factor receptor antibody appeared in cytoplasm, which further verified that OECs still survived and integrated well within the host 6 weeks after transplantation. Linear MBP positive fibers appeared in the injured region by MBP staining,which indicated that myelin-like substance appeared and drew closely in both ends of injured region. At the same time,MBP positive substance also appeared in the multipolar cells, which illustrated that transplanted OECs could induce the occurrence of myelin-like substance. No regenerated axon was found in the control group.CONCLUSION: Delayed transplantation of OECs can survive and induce the occurrence of myelin-like substance in injured spinal cord of adult rats.

Objective To investigate the clinical efficacy of drug-eluting stent( DES) combined with clopi-dogrel therapy in patients with coronary heart disease. Methods 123 patients underwent percutaneous coronary intervention (PCI) were randomly divided into observation group (DES) of 60 cases and the control group ( bare-metal stent,BMS) of 63 cases. The observation group was given the treatment of DES combined clopidogrel, the control group was given the treatment of BMS combined clopidogrel. The stent implantation and the follow-up two-year MACE were observed. Results (1) The average implanted stent length of the observation group was longer than that of the control group(P<(0.05) ,the angiography in vascular disease and complexity between the two groups had no significant difference ( P > 0. 05 ) ; the success rate of surgery in the two groups was 100% , after operation the TIMI flow reached grade 3. (2) The incidence of angina pectoris and revascularization rate of the observation group was significantly lower than that of the control group (P < 0.05); the rate of coronary angiography, recurrent myocardial infarction and the incidence of cardiac death between the two groups had no significant difference ( P > 0.05). Conclusion DES combined with clopidogrel maintenance therapy for a year in patients with coronary heart disease had a good long-term efficacy.

OBJECTIVE: To evaluate the effect of hypoxia improvement in Hep-2 cell on cisplatin-induced apoptosis. METHOD: Human laryngeal squamous cell carcinoma Hep-2 cells and HIF-1α-RNAi-Hep-2 cells were cultured in normoxic, hypoxic and reoxygenation condition. The inhibition of cisplatin on cell proliferation was evaluated by MTT assay. The influence of cisplatin on cell cycle and apoptosis were detected by flow cytometry. RESULT: The inhibition of cisplatin on cell proliferation was reduced by hypoxia. After HIF-1α gene was silenced, the inhibition of cisplatin on Hep-2 cell proliferation was increased apparently, but was still interfered partly by hypoxia. Hypoxia could induce cell apoptosis decreased and enhance chemotherapeutic resistance. After reoxygenation, cell apoptosis induced by cisplatin was increased significantly (P < 0.05). HIF-1α-RNAi-Hep-2 cells under hypoxia also showed certain resistance to apoptosis but the sensitivity to cisplatin was higher than that of Hep-2 cells. When cells were returned from hypoxic condition to normoxic condition for some time, the apoptosis induced by cisplatin was increased significantly (P < 0.05). CONCLUSION The improvement of hypoxic microenvironment with HIF-1α gene knockout could increase the sensitivity of Hep-2 cells to chemotherapy.
Apoptosis ; Carcinoma, Squamous Cell ; pathology ; Cell Cycle ; Cell Hypoxia ; Cell Line, Tumor ; drug effects ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; Flow Cytometry ; Gene Knockout Techniques ; Head and Neck Neoplasms ; pathology ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; Laryngeal Neoplasms ; pathology ; RNA Interference ; Squamous Cell Carcinoma of Head and Neck

Purpose Investigating the significance of ZO-1 different domains in the invasion and metastasis of gastric carcinoma (GC).Methods A tissue microarray that simulates the invasion and metastasis process of GC was created,and immunohistochemistry was performed to detect the expression of ZO-1 (α-pan),ZO-1 (α +) and ZO-1 (ZU5).Results The GC cell exhibited aberrant expression of ZO-1 (α-pan),ZO-1 (α +) and ZO-1 (ZU5) from membrane translocated to cytoplasm or no expression.The aberrant degree was increased with the invasion,however,was decreased in metastatic lymph node.The aberrant expression was associated with histological types.Conclusion The aberrant expression of ZO-1 (α-pan),ZO-1 (α +) and ZO-1 (ZU5),from membrane translocated to cytoplasm or no expression suggest that domains of PDZ3,GUK,SH3,ZU5 and alpha motif in ZO-1 might be involved in the invasion and metastasis of GC and maintaining of GC phenotype.The aberrant expression of these domains may be the one mechanism of ZO-1 involved in EMT or MET.

Objective To explore the diagnosis of cystinosis.Methods The clinical and biochemical information, and gene detection results in a child with cystinosis was retrospective analyzed.Results Four-year-old female presented with photophobia and corneal crystal was found by ophthalmic examination at 2 years old, bilateral kidney stone was found, accompanied by development delay and rickets at 3 years old. Gas chromatography analysis in urine showed that a variety of amino acids were increased, and urine sugar and urinary micro-protein were also increased, which were in accordance with fanconi syndrome. The blood free carnitine was decreased, ester acyl carnitine spectrum was normal, and multi-amino acids such as lysine, valine and arginine were decreased. Gene analysis showed a homozygous mutation of c.696C>G (p.323 N>K) inCTNS gene, which was a known mutation. Both her parents were carrier of heterozygous mutation of c.696C>G inCTNS gene.Conclusion Child with kidney stone, renal damage, combined by multi-system damage such as eyes, bone, and thyroid should be paid attention to identify the cystinosis.

OBJECTIVE: To observe the short-term efficacy of autogenous bone pate and Palva graft for obliterating huge remnant mastoid cavity in canal wall down approach. METHOD: Retrospective analysis clinical data of twenty-one cholesteatomatous cases operated by one surgeon from 2004 to 2007. In twelve cases, simultaneous III type tympanoplasty (Sheehy, P. O. P) was performed. Other 9 cases had undergone mastoidectomy elsewhere before the admission. Six of them were still draining with huge remnant mastoid cavity, and the rest three patients had relapsed cholesteatomas with intermittent draining and huge mastoid cavity. Normal saline solution perfusion was used to measure the volume of remnant mastoid cavity. The criterion of huge remnant mastoid cavity is more than 8 ml. RESULT: Of twelve primary cases with III type tympanoplasty, 11 patients maintained a small, dry, and healthy mastoid cavity after twenty-seven days. The average increase of hearing level of them was 17.5dB, and the air-bone gap is less than 20 dB. Of one patient, bone pate was infected and was discharged. A dry mastoid cavity was achieved until fifty-five days after surgery. The patient is keeping a big air-bone gap caused by displacement of ossicle chain prosthesis. Just eighteen days later, other nine cases of revision mastoidectomy achieved a small, dry, and healthy mastoid cavity, with lightly improved hearing level. CONCLUSION Obliteration of a canal wall down huge mastoid cavity by Palva graft with autologous bone pate is a reliable and effective technique that results in a small, dry, low-maintenance mastoid cavity. The short-term efficacy of simultaneous III tympanoplasty is satisfactory if patient selection is suitable.
Adolescent ; Adult ; Cholesteatoma ; surgery ; Cholesteatoma, Middle Ear ; surgery ; Female ; Humans ; Male ; Mastoid ; surgery ; Middle Aged ; Retrospective Studies ; Surgical Flaps ; Transplantation, Autologous ; Treatment Outcome ; Tympanoplasty ; methods ; Young Adult

Objective:To analyze the clinical features, genetic characteristics, treatment and follow-up results of patients with hydrocephalus caused by methylmalonic acidemia combined with homocysteinuria, and to discuss the optimal strategies for assessing and treating such patients.Methods:From January 1998 to December 2020, 76 patients with hydrocephalus due to methylmalonic acidemia combined with homocysteinuria in the Department of Pediatrics in 11 hospitals including Peking University First Hospital were diagnosed by biochemical, genetic analysis and brain imaging examination. The patients were divided into operation-group and non-operation-group according to whether they underwent ventriculoperitoneal shunt. The clinical features, laboratory examinations, genotype, and follow-up data were retrospectively analyzed. Data were compared between the two groups using rank sum test, and categorical data were compared using χ 2 test. Results:Among the 76 patients (51 male, 25 female), 5 were detected by newborn screening, while 71 were diagnosed after clinical onset, 68 cases (96%) had early-onset, 3 cases (4%) had late-onset. The most common clinical manifestations of 74 cases with complete data were psychomotor retardation in 74 cases (100%), visual impairment in 74 cases (100%), epilepsy in 44 cases (59%), anemia in 31 cases (42%), hypotonia or hypertonia in 21 cases (28%), feeding difficulties in 19 cases (26%) and disturbance of consciousness in 17 cases (23%). Genetic analysis was performed in 76 cases, all of whom had MMACHC gene variations, including 30 homozygous variations of MMACHC c.609G>A. The most common variations were c.609G>A (94, 62.7%), followed by c.658_660del (18, 12.0%), c.567dupT (9, 6.0%) and c.217C>T (8, 5.3%). Therapy including cobalamin intramuscular injection, L-carnitine and betaine were initiated immediately after diagnosis. A ventriculoperitoneal shunt operation was performed in 41 cases (operation group), and 31 patients improved after metabolic intervention (non-operation group). There was no significant difference in the age of onset, the age of diagnosis, the blood total homocysteine, methionine, and urinary methylmalonic acid concentration between the two groups (all P>0.05). The symptoms of psychomotor development, epilepsy, and visual impairments improved gradually after a long-term follow-up in the operation group. Conclusions:Hydrocephalus is a severe complication of methylmalonic acidemia combined with homocysteinuria. The most common clinical manifestations are psychomotor retardation, visual impairment, and epilepsy. It usually occurs in early-onset patients. Early diagnosis and etiological treatment are very important. Hydrocephalus may improve after metabolic intervention in some patients. For patients with severe ventricular dilatation, prompt surgical intervention can improve the prognosis.

Objective:To investigate the factors affecting phenotypes in the patients of methylmalonic acidemia combined with homocysteinemia cblC type with MMACHC c. 609G>A homologous variant. Methods:A retrospective study on the clinical manifestations, complications, treatment, and outcome in 164patients of cblC type with MMACHC c. 609G>A homologous variant was conducted.The patients were diagnosed by biochemical and genetic analysisfrom January 1998 to December 2020. Results:Among the 164 patients, 2 cases were prenatally diagnosed and began treatment after birth. They are 3 and 12 years old with normal physical and mental development. Twenty-one cases were diagnosed by newborn screening. Among them, 15 cases had with normal development. They were treated fromthe age of two weeks at the asymptomatic period. Six cases began treatment aged 1 to 3 months after onset. Their development was delayed. One hundred and forty-one cases were clinically diagnosed. Their onset age ranges from a few minutes after birth to 6 years old. 110 cases had early-onset (78.0%). 31 cases had late-onset (22.0%). Five of them died. 24 patients lost to follow-up. Of the 141 clinically diagnosed patients, 130 (92.2%) with psychomotor retardation, 69 (48.9%) with epilepsy, 39 (27.7%) with anemia, 30 (21.3%) had visual impairment, 27 (19.1%) had hydrocephalus, 26 (18.4%) had feeding difficulties, 7 (5.0%) with liver damage, and 5 (3.5%) with metabolic syndrome. The frequency of hydrocephalus and seizures was significantly higher in the early-onset group. The urinary methylmalonic acid increased significantly in the patients with epilepsy. During the long-term follow-up, the level of plasma total homocysteine in the seizure-uncontrolled group was significantly higher than that in the seizure-controlled group, the difference had a statistical significance ( P<0.05). Conclusion:Most of the patients with MMACHC c. 609G>A homozygous variant had early-onset disease, with a high mortality and disability rate. If not treated in time, it will lead to neurological damage, resulting in epilepsy, mental retardation, hydrocephalus, and multiple organ damage. Pre-symptomatic diagnosis and treatment are crucial to prevent irreversible neurological damage. Neonatal screening and prenatal diagnosis are important to improve the outcome of the patients.

Objectives:To summarize the clinical and genetic characteristics of the patients with isolated methylmalonic acidemia and investigate the strategies for the diagnosis, treatment and prevention.Methods:Three hundred and fourteen patients (180 males, 134 females) with isolated methylmalonic acidemia were ascertained from 26 provinces or cities across the mainland of China during January 1998 to March 2020. Genetic analysis was performed by Sanger sequencing, gene panel sequencing, whole exome sequencing, multiplex ligation-dependent probe amplification or quantitative PCR. According to the age of onset, the patients were divided to early-onset group (≤12 months of age) and the late-onset group (>12 months of age). They were treated by cobalamin, L-carnitine and (or) special diet and symptomatic treatment. Statistical analysis was done using Chi-square test.Results:Fifty-eight of 314 (18.5%) patients were detected by Newborn screening using liquid chromatography tandem mass spectrometry. Five cases (1.6%) had a postmortem diagnosis. Two hundred and fifty-one patients (79.9%) were clinically diagnosed with an age of onset ranged from 3 hours after birth to 18 years. One hundred and fifty-nine patients (71.0%) belonged to early-onset groups, 65 patients (29.0%) belonged to the late-onset group. The most common symptoms were metabolic crises, psychomotor retardation, epilepsy, anemia and multiple organ damage. Metabolic acidosis and anemia were more common in early-onset patients than that in late-onset patients (20.8%(33/159) vs. 9.2% (6/65), 34.6% (55/159) vs. 16.9% (11/165), χ 2=4.261, 6.930, P=0.039, 0.008). Genetic tests were performed for 236 patients (75.2%), 96.2%(227/236) had molecular confirmation. One hundred and twenty-seven variants were identified in seven genes (MMUT, MMAA, MMAB, MMADHC, SUCLG1, SUCLA2, and MCEE), of which 49 were novel. The mut type, caused by the deficiency of methylmalonyl-CoA mutase, was the most common ( n=211, 93%) cause of this condition. c.729_730insTT, c.1106G>A and c.914T>C were the three most frequent mutations in MMUT gene. The frequency of c.914T>C in early-onset patients was significantly higher than that in late-onset patients (8.3% (18/216) vs. 1.6% (1/64), χ 2=3.859, P=0.037). Metabolic crisis was more frequent in mut type than the other types (72.6% (114/157) vs. 3/13, χ 2=13.729, P=0.001),developmental delay and hypotonia were less frequent in mut type (38.2% (60/157) vs. 9/13, 25.5% (40/157) vs. 8/13, χ 2=4.789, 7.705, P=0.030, 0.006). Of the 58 patients identified by newborn screening, 44 patients (75.9%) who were treated from asymptomatic phase developed normally whereas 14 patients (24.1%) who received treatment after developing symptoms exhibited varying degrees of psychomotor retardation. Conclusions:The characteristics of phenotypes and genotypes among Chinese patients with isolated methylmalonic acidemia were analyzed. Expanded the mutation spectrum of the associated genes. Because of the complex clinical manifestations and severe early onset of isolated methylmalonic acidemia, Newborn screening is crucial for early diagnosis and improvement of prognosis. MMUT gene is recommended for carrier screening as an effort to move the test earlier as a part of the primary prevention of birth defects.