BACKGROUND: Nitric oxide (NO) is synthesized by catalysis of nitric oxide synthase (NOS). Three distinct isoforms of NOS have been detected in different structures of the guinea pig cochlea. However, there are still rather controversies about the definite localization and expression of NOS isoforms in guinea pig cochlea.OBJECTIVE: To investigate localization and expression of three NOS isoforms in the cochlea of guinea pigs, and to explore the effect of NO on auditory physiology and pathophysiology of inner ear.DESIGN: An observed and controlled study.SETTING: Department of Physiology, Jinzhou Medical College; Department of Orthopedics, the Second Affiliated Hospital of Jinzhou; Department of Otorhinolaryngology, Jinzhou Central Hospital.MATERIALS: The study was performed in Hearing Research Laboratory of China Medical University from May to November 2000. Ten healthy male albino guinea pigs, with body mass of 250-300 g, with sensitive Preyer's reflexes and normal drum membrane and external auditory canal,were selected.METHODS: After anesthesia, guinea pigs were decapitated and the temporal bones were removed immediately. The round and oval windows were opened, perfused with 40 g/L paraformaldehyde, and the specimens were immersed in the same fixative solution for 2 hours. Decalcification of the cochleae was performed in 100 g/L EDTA solution for 1 week. Then the specimens were placed in 250 g/L sucrose solution overnight and embedded in OCT. Cryostat (20 μm) sections were prepared for immunohistochemistry.MAIN OUTCOME MEASURES: Localization and expression of three NOS isoforms in the cochlea of guinea pigs.RESULTS: All data of ten guinea pigs was entered the final analysis without any loss. [1] Neuronal-type nNOS and endothelial-type eNOS immunoreactivity were localized in inner and outer hair cells of the organ of Corti, as well as in spiral ganglion cells. In addition, staining for nNOS and eNOS were also seen in the marginal cells of stria vascularis, spiral ligament cells, and in nerve fibers. [2] Inducible-type iNOS immunoreactivity was also detected in above each structure of the guinea pig cochlea under physiological conditions.CONCLUSION:NO may play an important role in neurotransmission,blood flow regulation, and cytotoxicity.

ThemagneticFe3O4nanoparticlesweresynthesizedbyco-precipitationmethod,andthenmagnetic Fe3 O4@Au nanoparticle was synthesized to improve the affinity of particle surface. L-Cys-GA3 was grafted on the surface of gold clad by self-assembly method, and then dropped it on glassy carbon electrode, for further manufacture of MIP/Fe3 O4@Au by using electropolymerzation L-Cys. The surface morphology and particle size distribution of Fe3 O4@Au were studied by TEM. The structure and composition of gibberellins A3, MIP and nMIP were studied by IR. The test system was optimized, and the results showed that when the cycles of electropolymerization was 30, acetic acid:methanol (1:8, V/V) was chosen as eluent, elution time was optimized for 5 min and rebinding time for 7 min, the sensor got a high stability and good recognition ability for gibberellins A3 . The concentration of gibberellins A3 in the range of 1 . 0 × 10-11-1 . 0 × 10-8 mol/L had a relationship with the oxidation peak current of probe, with the detection limit of 2. 57×10-12 mol/L. The sensor was successfully used for the determination of GA3 in beer sample.

In order to solve the problems of present quartz crystal microbalance ( QCM ) measuring instruments, such as high demand for crystal cutting technology and uncomprehensive measurements for crystal parameters, a modified quadrature demodulation-based method was proposed with broadband adaptive response capacity and high frequency resolution. Moreover, it is also capable of measuring both resonant frequencies and dissipation factor D synchronously and continuously. Experimental results at room temperature indicated that the adaptive frequency range was 1-9 MHz while the frequency resolution was less than 1 Hz, measured resonance frequency shifts of crystals in the range scale linearly with the equal thickness increments of poly acrylic acid ( PAA) membrane on the working electrode, and with the volatilization of different solvent factor D is measured continuously and effectively along the time axis. To sum up, compared with traditional ones, this new method has lower material cost and more obtained parameters.

Objective To investigate the influence of the lymph node micrometastasis and its clinicopathological features on postoperative disease-free survival rate for patients with gastric cancer.Methods The study included 120 patients with pT1-3NoMo gastric cancer. The relationships between clinicopathological features or carcinoembryonic antigen (CEA) positive expression and postoperative disease-free survival rate were analyzed. Results In clinicopathological factors, multivariate analysis identified CEA positive expression was significantly correlated with tumor diameter (P = 0.011 ),depth of tumor invasion (P= 0.027) and lymphatic vessel invasion (P= 0.001 ) in lymph node positively. The average postoperative follow-up was (53.14 ± 16.75) months. There was statistical correlation between the tumor diameter( P = 0.018 ) or depth of tumor invasion ( P = 0.015 ) and postoperative disease-free survival rate. The disease-free survival rate was 90.91% ( 80/88 ), 86.36% ( 19/22 )and 40.00% (4/10) for the lymph node CEA negative,isolated tumor cells (IT Cs) and micrometastasis,respectively. There was significant difference between micrometastasis and the lymph node CEA negative (P= 0.000) or ITCs (P = 0.009), however, the lymph node CEA negative and ITCs was no significant difference (P = 0.438 ). Lymph node micrometastssis of gastric cancer was detected in 10 patients who should belong to stage pN1,the restage rate was 8.33%(10/120). Conclusions If the patients were found micrometastasis in lymph node with high-risk stage pT1-3NoMo gastric cancer for whom chemotherapy may be recommended,because of its high recurrence and poor prognosis.

Objective: To investigate the nutritious effects of IGF, HGF and Gln on intestinal mucosal precursor cells in vitro.Methods: Different combinations of IGF,HGF and Gln were used to observe their effects on the expansion and development of the cultured intestinal mucosal precursor cells.Results: The combination of IGF,HGF and Gln could markedly promote the expansion of cultured intestinal mucosal precursor cells.Although IGF or HGF alone could promote the expansion of intestinal mucosal precursor cells, they had little effects on the development.On the contrary,Gln alone can promote the development of intestinal mucosal precursor cells,but it had little effects on their expansion.Conclusion: IGF,HGF or Gln alone has little effects on the expansion and development of the intestinal mucosal precursor cells.When they are used together,they can efficiently provide the essential stimuli for the expansion of the cultured intestinal mucosal precursor cells in vitro and enable the recombination of the different cells and formation of intestinal mucosa-like structure in vitro.

OBJECTIVE: To analyze gene variants in a Chinese pedigree with oculocutaneous albinism (OCA). METHODS: Gene sequencing of the proband and his parents was performed using chip capture high-throughput sequencing and Sanger sequencing techniques, and PolyPhen-2, SIFT, MutationTaster, and FATHMM software were used to predict the function of new variants. At the same time,the pedigree and variant genes of 4 albinism patients from this pedigree were analyzed. RESULTS: Sequencing results showed that the proband's TYR gene (NM_000372) has c.230G>A (p.Arg77Gln) and c.120_121insG (p.Asp42GlyfsTer35) compound heterozygous variants. The proband's father carries c.230G>A heterozygous variant, and the mother carries c.120_121insG heterozygous variant, indicating that the proband's two variants are from his father and mother. The former is a known missense variant, which can cause abnormal or loss of the original function of the protein polypeptide chain. The latter c.120_121insG(p.Asp42GlyfsTer35) is an unreported frameshift variant of the TYR gene subregion (EX1; CDS1). PolyPhen-2, SIFT, MutationTaster and FATHMM predictions are all prompted as "harmful variants". This variant caused the amino acid encoded protein to terminate prematurely, producing a truncated protein, which eventually formed a 76-amino acid short-type TYR protein instead of the 529-amino acid wild-type TYR protein. Through the pedigree analysis, the four patients in the pedigree are all of the same type of compound heterozygous variants, and the disease-causing genes are all from the patient's parents. They belong to a special form of consanguineous marriage within 5 generations. CONCLUSION The compound heterozygous variants of c.230G>A (p.Arg77Gln) and c.120_121insG (p.Asp42GlyfsTer35) of the TYR gene may underlie the disease in this pedigree. The gene sequencing results enrich the variant spectrum of the TYR gene, and has facilitated molecular diagnosis for the patient.
Albinism, Oculocutaneous/genetics* ; Consanguinity ; Heterozygote ; Humans ; Mutation ; Pedigree

Objective:To observe the effects of hyperthermia on the sensitivity of gastric cancer cell lines to paclitaxel (TAX) . Methods :To establish TAX-resistant gastric cancer cell line through a method of increasing concertration gradient and named it MKN-45/TAX .Immunocytohistochemistry staining method was used to detect the expression of multi-drug resistance gene (MDR1) in MKN-45/TAX and MKN-45 .CCK-8 test was used to detect the proliferation inhibition rates in two kinds of cell lines under different temperatures and different TAX concentration .Real-time polymerase chain reaction (RT-PCR) and West-ern blotting were used to detect the expression of MDR1 mRNA and P-glycoprotein (P-gp) before and after target siRNA treat-ment .Results:Establishment of MKN-45/TAX cell line was successful .The expression level of MDR1 was high in MKN-45/TAX whereas it was low in MKN-45 .As TAX concentration increased ,the proliferation inhibition rates of both cell lines in-creased .At 42 ℃ the proliferation inhibition rate of MKN-45/TAX to TAX chemotherapy decreased ,while that of MKN-45 was the opposite .Both the expression of MDR1 mRNA and P-gp decreased after the siRNA transfection .Conclusions :Hyper-thermia combined with chemotherapy may enhance the resistance of TAX-resistant gastric cancer cell line to TAX ,while it may enhance the sensitivity of TAX-sensitive gastric cancer cell line to TAX .The mechanism may be related to the expression of MDR1 .

Objective:To compare the clinical efficacy of laparoscopy-assisted choledochoelectroscopy and endoscopic retrograde cholangiopancreatography on treatment of acute cholangitis with bile duct stones .Methods :The clinical datum of 103 cases of a-cute cholangitis with bile duct stones treated by laparoscopy-assisted choledochoelectroscopy (Group L ,n=64) or endoscopic retrograde cholangiopancreatography (Group E ,n=39) from October 2008 to September 2013 were analyzed retrospectively . In hospital days ,costs ,the rates of residual stones ,the rates of complications and the stone recurrence rates were compared between the two groups .Results:The operation success rates were 96 .25% and 89 .74% ,and hospital days were (9 .6 ± 0 .9) and (8 .3 ± 0 .7) days in Group L and Group E ,respectively ,but had no stastically significant differences (P> 0 .05) .The cost ,the rate of residual stones ,the rates of complications and the stone recurrence rate in Group L were significantly lower than those in Group E (P<0 .05) .Conclusions :Acute cholangitis with bile duct stones can be treated by laparoscopy-assisted choledochoelectroscopy or endoscopic retrograde cholangiopancreatography .However ,the former presents more clinical advan-tages .

OBJECTIVETo extract and purify of solamargine from Solanum nigrum, and to research its antineoplastic effects.

METHODS. nigrum was extracted refluently with 80% alcohol, solamargine was purified with silica gel column chromatography and recrystallization, and then conducted its structure identification and purity checks. Screened the effect on human tumor cell groth inhibition in vitro by MTT assay, and researched on the features in mice with H22 liver cancer or Ehrlich ascites tumor of solamargine.

RESULTThe concent of solamargine reached 97.9%. Solamargine had significantly inhibition on 6 tumor cells in vitro, and it had significantly inhibition on mice with H22 liver cancer or ehrlich ascites tumor in the 2.4 mg x kg(-1) dose of i.v.

CONCLUSIONSolamargine have the antineoplastic effect.

Animals ; Antineoplastic Agents, Phytogenic ; isolation & purification ; Cell Line, Tumor ; Female ; Humans ; Mice ; Mice, Inbred ICR ; Solanaceous Alkaloids ; isolation & purification ; pharmacology

OBJECTIVETo explore the effects of the method of soothing the liver and regulating qi on expression of gastrin and somatostatin in hypothalamus and gastric antrum of functional dyspepsia model rats.

METHODThe 32 rats were randomly divided into normal group, model group, Chaihu Shugansan group and domperidone group (n = 8). The functional dyspepsia model was established by constantly squeezing their tails and mean while saline, Chaihu Shugansan decoction and domperidone suspension were administered respectively to 4 groups by gavage. The expression of gastrin and somatostatin in hypothalamus and gastric antrum of rats by immunohistochemical were detected 3 weeks later.

RESULTThe expression of GAS in the hypothalamus and gastric antrum of model group were less than those of normal group (P < 0.05, P < 0.01), while the expression of SS in the hypothalamus and gastric antrum in Model group were significantly increased than those of normal group (P < 0.01). The expression of GAS and SS in gastric antrum of Chaihu Shugansan group and domperidone group were increased and decreased respectively, and the differences were significant (P < 0.05, P < 0.01). There were no obvious difference about expression of GAS, SS in the hypothalamus between domperidone group and model group. GAS expression in hypothalamus of Chaihu Shugansan group were increased than those of normal group but there was no obvious difference in SS expression in hypothalamus between Chaihu Shugansan group and model group.

CONCLUSIONThe method of soothing the liver and regulating qi can increase GAS expression in central and peripheral and decrease SS expression in peripheral gastric antrum, which may be one of its therapeutic mechanisms on functional dyspepsia.

Animals ; Disease Models, Animal ; Drugs, Chinese Herbal ; therapeutic use ; Dyspepsia ; drug therapy ; genetics ; metabolism ; Female ; Gastrins ; genetics ; metabolism ; Gene Expression Regulation ; drug effects ; Humans ; Hypothalamus ; drug effects ; metabolism ; Liver ; drug effects ; physiopathology ; Pyloric Antrum ; drug effects ; metabolism ; Qi ; Random Allocation ; Rats ; Rats, Wistar ; Somatostatin ; genetics ; metabolism