Objective:To establish an HPLC method for the simultaneous determination of methionine, vitamin B1 and vitamin B2 in Lingzhi Erwei methionine capsules. Methods:The determination was performed on a Ultimate? AQ-C18 column(250 mm × 4. 6 mm, 5 μm,Welch Inc. ) with the detection wavelength at 220 nm. The mobile phase was composed of methanol and 0. 07 mol·L-1 sodium heptanesulfonate solution (14 ml triethylamine diluted with water to1 000ml, adjusting pH to 3. 5 with diluted orthophosphoric acid) with gradient elution, and the flow rate was 1. 0 ml·min-1. Results: Methionine, vitamin B1 and vitamin B2 was in good linearity within the range of 0.527 0-1.510 0 mg·ml-1 - (r =0.999 9), 0.045 52-0.136 56 mg·ml-1 (r =0.999 9) and 0.010 10-0. 060 58 mg·ml-1(r=0. 999 9), respectively. The mean recovery was 100. 5%, 97. 7% and 101. 5% with RSD of 0. 5%, 1. 0%and 1. 4%(n=9) accordingly. Conclusion:The method is simple, accurate, reliable and appropriate in the simultaneous determina-tion of methionine,vitamin B1 and vitamin B2 in Lingzhi Erwei methionine capsules.

Objective To investigate the clinical character and treatment method of the coexistence of cervical and lumbar spinal stenosis.Method Twenty-seven cases with coexistence of cervical and lum bar spinal stenosis,9 cases with single cervical operation,7 cases with single lumbar operation,11 combined cervical and lumbar operation.Results Twenty-two cases were followed-up,average 49 months,all eases were recorded with JOA score.JOA score was(8.6±0.2)scores preoperafion,(11.7±0.3)scores 2 weeks postoperation,(13.1±0.2)scores 3 months postoperation,(13.5±0.2)scores 6 months postoperation,(13.9±0.3)scores 12 months postoperation.Relief rate was 79.2%,satisfactory rate was 93.8%.Conclusions The patient with multi-segment spinal stenosis should be treated individually,to certain responsible focus.If no responsible focus,it is better to treat with cervical spinal operation first.

OBJECTIVETo observe the expressions of Calbindin(CB) and Parvalbumin (PV), the two calcium-binding protein, in auditory pathway in mice of wild type C57BL/6J and kit⁺/kitW⁻ ²Bao, a kit gene mutant.

METHODSSix mutated kit gene kit⁺/kitW⁻ ²Bao mice and 6 wild type C57BL/6J (B6) mice were anaesthetized i. p. with chloral hydrate. After the mice were fixed by heart perfusion, the brains were removed and coronal sections were cut with a freezing microtome.

RESULTSWe found that wild type mice had significant expressions of PV on ventral cochlear nucleus, anterior part (AVCN), ventral cochlear nucleus, posterior part (PVCN), inferior colliculus (IC) and auditory cortex (AC). CB was expressed in wild type mice on PVCN and nucleus of the trapezoid body (Tz). The mutant of kit gene induced the less expression of PV on PVCN, IC and AC (P < 0.01), but increased the expression of Tz (P < 0.01). CB could not be observed on PVCN in mutant mice, and the expression of AC was increased( P < 0.01).

CONCLUSIONCB and PV has differential expression level in auditory pathway. Since mutated kit gene can affect expression of PV on PVCN, IC, Tz and AC, as well as CB on PVCN and AC, it suggests that the mutation of kit gene can affect the advanced function of central nervous system in auditory pathway.

Animals ; Auditory Cortex ; metabolism ; Auditory Pathways ; metabolism ; Calbindins ; metabolism ; Inferior Colliculi ; metabolism ; Mice ; Mice, Inbred C57BL ; Mutation ; Parvalbumins ; metabolism ; Pons ; metabolism ; Proto-Oncogene Proteins c-kit ; genetics

Objective To investigate the clinical value of color Doppler ultrasound in the early diagnosis of acute cholecystitis infection ,in order to improve the diagnosis of acute cholecystitis infection .Methods 36 cases with acute cholecystitis infection were selected as the observation group , and during the same period the healthy people were randomly selected as the control group .The color Doppler performance of ultrasound was observed .The hepatic artery blood flow velocity and the maximum peak of the cystic artery (Vmax) and resistance index(RI) were meas-ured.ROC curve was observed to evaluate the specificity of color Doppler ultrasound for the disease .Results The incidence rates of weak bile sound transmission ,gallbladder wall thickness ,gallbladder enlargement ,ultrasound Mur-phy positive in the control group were 25.0%,16.7%,22.2%,8.3%,which in the observation group were 72.2%, 47.2%,100.0%,94.4%,the differences were statistically significant (χ2 =4.571,4.571,4.589,5.689,all P<0.05).The differences in hepatic artery and the cystic artery Vmax and RI were statistically significant between the two groups(P<0.05).ROC curve comparison showed that the correct rate of gallbladder enlargement in the observa -tion group was the highest (90.7%),the specificity was 90.7%,sensitivity was 90.6%,and other indicators of ultra-sound had no significant difference (χ2 =0.893,P>0.05).Conclusion Color Doppler ultrasound can show the form of acute infectious cholecystitis ,gallbladder wall degree of bile through sound etc .,it can improve the diagnostic value of the disease.

Objective To monitor the antibiotic resistance in Neisseria gonorrhoeae, determine the prevalence of penicillinase-producing Neisseria gonorrhoeae (PPNG) and plasmid-mediated tetracycline-resistant Neisseria gonorrhoeae (TRNG) in Xi'an region, and to analyse the trends in antibiotic resistance in Neisseria gonorrhoeae. Methods In total, 647 strains of Neisseria gonorrhoeae were isolated from patients with gonorrhea in sexually transmitted disease (STD) clinic settings from 2002 to 2009. Agar dilution method was used to detect TRNG and determine the minimal inhibitory concentration (MIC) of antibiotics, and paper acidometric method to detect PPNG. Results Of these 647 strains, 216 (33.4%) were TRNG, 290 (44.8%)were PPNG. The prevalence of TRNG strains remained between 28.3% and 49.2% in 2002-2009, except for 17.3% in 2005; the prevalence of PPNG strains increased from 37.1% in 2002 to 64% in 2005, but declined from 2006 to 2009 (32.3%). The prevalence of resistance to spectinomycin maintained at a low level (0 to 2.8%) over these years, while that to ciprofloxacin remained higher than 80% from 2002 to 2009, and accounted for 100% in 2005, with the exception of 51% in 2006. Ceftriaxone resistance was observed in none of these strains except 4 isolates in 2003, but the susceptibility to ceftriaxone decreased yearly. Conclusions Neisseria gonorrhoeae is highly sensitive to spectinomycin, which should serve as the first treatment choice for gonorrhea.Full dose is necessary for the application of ceftriaxone in the treatment of gonorrhea. Ciprofloxacin should not be used to treat gonorrhea.

BACKGROUND: It has been demonstrated that insulin-like growth factor-1 (IGF-1) is a kind of neurotrophic factor and protects from cerebral ischemia/reperfusion injury, the expression of IGF-1 is associated with the attack of ischemic stroke. The effects of IGF-1 and its receptor (IGF-1R) on neurobehavioral function are to be further studied.OBJECTIVE: To observe the effects of IGF-1 and IGF-1R on neurobehavioral function in rat models of cerebral ischemia/reperfusion injury.DESIGN: A randomized controlled observation.SETTING: Institute of Cerebrovascular Diseases, Affiliated Hospital of Qingdao University Medical College.MATERIALS: The experiments were carried out in Shandong Key Laboratory for Prevention and Treatment of Brain diseases. Twenty-eight healthy adult Wistar rats of clean degree, weighing 220-260 g, were provided by the experimental animal center of Shandong University.METHODS: The rats were randomly divided into experimental group (n =24) and sham-operated group (n =4). The middle cerebral artery occlusion/reperfusion (MCAO/R) models were established by inserting a thread through left external-internal carotid arteries. The sham-operated rats were given the same treatments except inserting thread. ①Neurologic deficit test: The rats in the experimental group were assessed according to Bederson standard after 1-hour ischemia and 6, 12-hour, 1, 3, 7 and 14-day reperfusion respectively. The sham-operated rats were assessed at corresponding time points; Without neurologic deficit was marked as 0 point; flexion of anterior claws as 1 point; unable to act against the pushing from the contralateral side as 2 points; circling while walking as 3 points; shaking as 4 points;unconscious mind as 5 points. ② Sample collection and treatment: The samples in the experimental group were collected after 1-hour ischemia and 6, 12-hour, 1, 3, 7 and 14-day reperfusion, and those in the sham-operated group ere collected at 24 hours postoperatively. The rats were anesthetized, brain samples were got at about 5 mm posterior to optic chiasma after brains were removed completely, then serial coronal sections (5 μm) were prepared, and 1 from 10 sections was stuck to the cover glasses treated with poly-L-lysine. ③ Morphological observation of neurons: The neurons in brain were observed by toluidine blue staining. ④ Detection of IGF-1 and IGF-1R: The expressions of IGF-1 and IGF-1R in cortex and striatum were detected with immunohistochemical technique, 4 fields were randomly selected to count the positive cells under high-power microscope (×400).MAIN OUTCOME MEASURES: ① The neurologic deficit; ② Morphological changes of neurons in brain; ③ Expressions of IGF-1 and IGF-1R in cortex and striatum.RESULTS: All the 28 rats were involved in the analysis of results. ① The neurologic deficit: The scores of neurologic deficit were (1.50±058) and (1.50±0.78) in rats after 7 and 14-day reperfusion, which were lower than that in rats after 6-hour reperfusion [(3.00±0.00), P ＜ 0.05]. ② Morphological changes of neurons in brain: The neurons in ischemic area appeared as paryopyknosis and became irregular in shape, there were obvious gaps around the cells, also deeply stained as purplish blue, nucleolus disappeared, and there were many scattered cellular fragments. ③ Expressions of IGF-1 and IGF-1R in cortex and striatum: The numbers of IGF-1 positive cells in cortex were (8.75±2.06), (11.13±1.14),(19.75±3.18), (17.38±3.11 ) and (11.23±2.28) respectively in rats after 6, 12-hours and 1, 3, 7-day reperfusion, which all were higher than that in sham-operated rats [(3.88±1.46), P ＜ 0.05], the numbers of IGF-1 positive cells in striatum were(8.25±2.21), (11.34±2.21), (18.23±2.64), (18.56±2.34) and (11.31±2.14) respectively in rats after 6, 12 hours and 1, 3, 7days reperfusion , which were also higher than that in sham-operated rats [(4.12±2.24), P ＜ 0.05]. The numbers of IGF-1R positive cells in cortex were (7.63±1.50), (10.50±2.34), (15.55±3.12), (15.37±3.01), (8.86±2.75) respectively in rats after 6, 12-hours and 1,3,7-day reperfusion, which all were higher than that in sham-operated rats [(4.13±1.81), P ＜0.05]. Those in striatum were (8.33±2.31), (10.24±2.09), (14.72±2.17), (14.24±2.77), (8.38±2.05), which were also higher than that in sham-operated rats [(3.76±2.35), P ＜ 0.05].CONCLUSION: The neurological function is damaged after cerebral ischemia/reperfusion, but it has a trend of self-recovery. The expressions of IGF-1 and IGF-1R are mainly distributed in cortex and striatum. Higher expressions of IGF-1 and IGF-1R maintain during 12 hours to 7 days after reperfusion and have a peak value at 1-3 days, which suggests that early expression of IGF-1 and IGF-1R are certain related to the recovery of neurological function.

BACKGROUND: Brain injury can induce the increased expression of basic fibroblast growth factor (bFGF) in brain,whereas FGFR is a very important player in the cell proliferation and differentiation, angiogenesis, skeletogeny, etc.OBJECTIVE: To observe the effect of bFGF and its receptor on neuronal apoptosis following cerebral ischemia/reperfusion injury in rats.DESIGN: A randomized grouping design and animal experiment.SETTING: Institute of Cerebrovascular Disease, Affiliated Hospital of Qingdao University Medical College.MATERIALS: Twenty-eight healthy adult Wistar rats of clean degree, weighing 220-260 g, were provided by the experimental animal center of Shandong University. Rabbit-anti-rat bFGF and fibroblast growth factor receptor-1(FGFR-1) monoclonal antibodies were provided by Wuhan Boster Biological Technology, Co.,Ltd.METHODS: The experiment was carried out in Shandong Key Laboratory for Prevention and Treatment of Brain diseases.① The rats were randomly divided into experimental group (n =24) and sham-operated group (n =4). Models of middle cerebral artery occlusion/reperfusion (MCAO/R) were established by thread occlusion via left external-internal carotid arteries, and 4 rats in the experimental group were sampled at 1-hour ischemia/6, 12-hour, 1, 3, 7 and 14-day reperfusion respectively. The rats in the sham-operated group were given the same treatment without inserting thread.After anesthesia, the brain was removed completely by cutting head, then the brain tissue at about 5 mm posterior to optic chiasma was cut down, then serial coronal sections (5 μm) were prepared. ② The brain tissues were stained with ematoxylin-eosin (HE), and the forms of neurons were observed under microscope. ③ TdT-mediated dUTP-biotin nick end labeling (TUNEL) method: there were buffy granules in nucleus which was positively stained (apoptosis). Four fields were randomly selected from cortex and striatum to count positive cells under high-power microscope (×400). ④ The sections were stained with rabbit-anti-rat bFGF and FGFR-1 monoclonal antibodies, 4 fields were randomly selected from cortex and striatum to count positive cells under high-power microscope (×400).MAIN OUTCOME MEASURES: Apoptosis and the expressions of bFGF and FGFR-1.RESULTS: All the 28 rats were involved in the analysis of results. ① In the experimental group, the neurons in the ischemic sites were obviously decreased, some neurons appeared as paryopyknosis and became irregular, also deeply stained as purplish blue, nucleolus disappeared, and there were many scattered cellular fragments. ② In the sham-operated group, there were a few apoptotic neurons in the brain tissue, and the apoptotic neurons were obviously increased after ischemia, which mainly observed in cortexes and striatums of frontal and paritetal lobes. In the experimental group, apoptotic cells in cortexes began to increase gradually at 6 hours, and there were more cells at 12hours and 3 days, which reached the peak value at 1 day, and began to decrease at 3 day, but there were still more apoptotic cells at 14 days than in the sham-operated group. The number of apoptotic neurons and the changing trend in striatums were generally the same as those in cortexes (P ＞ 0.05). ③ In the sham-operated group, there were weak bFGF expression in the neurons of brain tissue, but there were fewer lightly stained positive cells. After cerebral ischemia, the bFGF expressions were increased, mainly observed in cortexes and striatums. The bFGF expression appeared at 6 hours after cerebral ischemia/reperfusion, and the number was increased gradually and deeply stained as the time of reperfusion prolonged (Figure 3), it reached the peak value at 1-3 days, and then weakened gradually, but it was still higher than in the sham-operated group at 14 days [(5.01 ±1.71), (5.21 ± 1.62) cells/visual field; (2.03± 1.73),(2.46± 1.38) cells/visual field, P ＜ 0.05]. ④ In the sham-operated group, lightly stained FGFR-1 positive cells could be observed in brain tissue. At 6 hours after cerebral ischemia/reperfusion, the FGFR positive cells began to increased in cortexes and striatums, which were the most at 1-3 days, and gradually decreased after 3 days, and the number was still a little more than that in the sham-operated group at 14 days [(5.01± 1.41), (5.20± 1.33) cells/visual field; (2.25±1.67),(2.32± 1.61 ) cells/visual field].CONCLUSION: After cerebral ischemia/reperfusion, the expressions of endogenous bFGF and FGFR-1 may be activated in cortex and striatum, then inhibit the neuronal apoptosis, and play its neuroprotective role.

Objective To explore the generalization effect of a semantic navigation strategy(SNS)on naming in patients with aphasia.Methods Fourteen patients with aphasia were randomly divided into an SNS group (n =7)and an unrelated semantic(US)group(n =7).One hundred and twenty-two words chosen from Chinese word association norms were used in this study.The presentation sequence of words was generated automatically with network analysis techniques for the SNS group and randomly without network analysis techniques for the US group.After baseline tests using the western aphasia battery(WAB)and the mini-mental status examination(MMSE),all patients received language training once a day for 10 consecutive days.The amount of correct responses was recorded every day in order to draw the learning curve.The generalization effect was evaluated with 46 untrained items before and after training with all patients.Results The average learning curve of the SNS group was steeper than that of the US group.Correct responses on trained items were significantly more frequent in the SNS group than in the US group after the 6th day of training.After 10 days the frequency of correct responses on untrained items in the SNS group was significantly higher than it had been before training,and higher than the frequency after training in the US group.The frequency of no response on untrained items in the SNS group had declined,and it had declined more than in the US group.Conclusion A semantic navigation strategy may promote significant generalization while improving the learning curve in naming among patients with aphasia.

Objective To investigate the protective effect of GSH on lung injury in rats with severe acute pancreatitis(SAP) and investigate the mechanism of severe acute pancreatitis associated lung injury. Methods Fifty-four Spragwe Dawley rats were randomly divided into three groups: control group, model(SAP)group and herbal group. Rats in model group and herbal group were induced pancreatitis by retrograde injection of 5% sodium taurocholate into the pancreatobiliary duct. Animals were killed at 3h, 6h, 12h after injection. The serum amylase was examined at 3h, 6h, 12h after injection. Pulmonary tissue fluid content was measured.The expression of pulmonary TNF-α mRNA and LIF mRNA was detected by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Results The serum level of amylase was lowered significantly in herbal group in comparision with that in model group (P<0.05) and pulmonary tissue fluid content in herbal group was reduced compared with model group (P<0.05). Similarly, the level of pulmonary TNF-α mRNA and LIF mRNA expressions was both significantly reduced in herbal group compared with model group (P<0.05). LIF mRNA was not examined in normal control group. Conclusion Treatment with GSH can prevent lung injury of rats with SAP.

Sudden sensorineural hearing loss (SSHL),which is a common and frequently encountered disease,is considered to be a medical emergency in otolaryngology.The prevalence of SSHL is increasing in China.The pathogenesis of SSHL is not clear yet.Microcirculatory disorder of inner ear is considered as one of the most important causes of SSHL.In recent years,several reports have found the levels of serum lipids were changed in patients affected by SSHL.The relationship between SSHL and serum lipids was reviewed to provide new ideas for the diagnosis and treatment of SSHL.