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 effect of plasmid pEGFP- hepatocyte growth factor (HGF)-C1 on rat acute ischemia of hindlimb.Methods The eukaryotic expressed plasmid pEGFPHGF-C1 carrying human HGF cDNA was constructed.The transfection efficiency and the expression level of HGF were evaluated by transfecting pEGFP-HGF-C1 into primary rat skeletal muscle cells.Ligation-induced ischemia of femoral artery of one hindlimb in Wistar rats was performed.The plasmids (200 μg/500 μl) were injected once directly into the ischemic limb muscle (5 sites around ligation position) immediately after ligation.HGF expression was detected in the muscle tissue of rats on days 3,6 and 9 by immunochemical stain and Western blot.At day 24,the muscles were removed and stained with CD31 to assess histologically the capillary formation.Results The primary rat skeletal muscle cells could be transfected 0efficiently with pEGFP-HGF-C1 using LipofectamineTM2000 (0.8％),and secreted HGF which peak concentration was (5402.0±227.9) ng/L at 4 d.HGF expression was detected clearly in muscle tissue on days 3,6,9 in pEGFP-HGF-C1 groups.Western blot semi-quantitied analysis showed the levels of HGF expression were higher than control groups on days 3,6,9(P＜0.05).A significant increase in capillary density was found in rats transfected with human HGF(10.81±2.35) as compared with sham group (6.11±0.90) and control group (5.45±0.90) at 24 d(P＜0.01).Conclusions Intramuscular injection of naked human HGF plasmid may induce therapeutic angiogenesis in rat ischemic hindlimb models,as potential therapy for peripheral arterial disease.

The objective of the present study was to establish a novel rapid fluorescence focus inhibition test (RFFIT GFP)for the detection of rabies virus antibody, in which a chimeric rabies virus expressing green fluorescent protein (HEP GFP) was used as the basic virus strain in RFFIT GFP assay, and a few serum samples from human, dog and cat were detected by this new method .The optimal serum dilution, virus dosage and infection time were determined in 24 serum samples from human, dog and cat by using RFFIT GFP, RFFIT and ELISA assays. The result showed that these 3 methods gave a good consistency. But RFFIT GFP was found to be more convenient and economic for the detection of rabies virus antibody.

Objective To discuss the value of medium resolution typing method for HLA-B antigens of Northern Chinese by DNA chip technique.Methods The chip was made with specific medium distinguish typing probes designed according to gene frequency of HLA-B alleles from Northern Chinese.Unsymmetrical PCR was used to amplify HLA-B exon 2 and 3, then labeled PCR products and hybridize with probes on the chip.Certified the typing of HLA-B by analysed and scanned the signals of hybridize through a set of computer software.Results HLA-B alleles were successfully typed in 30 clinical samples. This medium distinguish probes were able to discern 42 HLA-B alleles from the scope of HLA-B 7~83. Using it we can distinguish B14,73 and 82 three new alleles contrast to PCR-SSP methods.Conclusions DNA typing of HLA-B by chip was proven to be a high-resolution and high-specificity method. It is able to check out the multitudinous samples in one DNA chip and it is more suitable for clinical application.

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.

The starting material L-tryptophan reacted with 4-chlorobenzaldehyde via Pictet-Spengler condensation and followed by oxidation and decarboxylation to afford the 1-(4-chlorophenyl)-beta-carboline. The intermediate was further reacted with alkyl halogenide by N(9)-alkylation and N2-quaternarization to obtain 12 novel 1-(4-chlorophenyl)-beta-carboline derivatives. The chemical structures of all target compounds were characterized by elemental analyses, MS and 1H NMR spectra. The antitumor activities of the target compounds were evaluated by MTT method. The results demonstrated that N2-quaternarized compounds enhanced the antitumor activity significantly. In particular, compound 15 was found to be the most potent compound with IC50 values lower than 5 micromol x L(-1) against 6 human tumor cells. These results confirmed that the N2-alkyl or aralkyl substituent on the beta-carboline ring played an important role in the modulation of the antitumor activities.

Objective:To establish a DNA typing method for HLA-I antigens with medium resolution method by DNA chip technique.Methods:The chip was made with specific medium distinguish-typing probes designed according to gene frequency of HLA-I alleles from Northern Chinese. Unsymmetrical PCR was used to amplify HLA-I exon2,3,and then the PCR products labeled and hybridized with probes on the chip.Typing of HLA-I was certified by scanning the hybridizing signals of through a set of computer software.Results:HLA-I alleles were successfully typed in 30 clinical samples .This medium-distinguishing probes were able to discern 57 HLA-I alleles accurately.Conclusion:DNA typing of HLA-I by chip has been proven to be a high-resolution and high-specific method. It is able to check out the new alleles that can not be distinguished by other methods with the same resolution., and it is more intuitional and more suitable for clinical application .

One strain of Pseudoalteromonas sp. E18 was isolated from the sea mud of Ningbo, Zhejiang. It can produce indigo pigment. The morphological, cultural and biochemical characteristics of the bacterium were studied. The indigo pigment was also extracted. The results showed that the maximum absorption peak of the pigment was at 579nm. The pigment was stable to UV, Na_2SO_3,It was also stable at pH 3～9. The pigment was unstable to the sunlight and high concentration H_2O_2. Temperature of 60℃～80℃ could increase the hue while temperature higher than 90℃ could reduce the hue.

Objective: To clarify the contribution of inhibitory effect of the chemical composition of fingerprint characteristic peaks from different parts of Lepdium meyenii (Maca) on PDE5, and to elucidate the material basis. Methods: The HPLC fingerprints of different parts from L. meyenii were established. The isotope labelling method was adopted to test the inhibitory rate of different extracted parts on PDE5.The gray relative analysis and partial least-squares method were used to make correspond analysis of the spectrum-effect relationship. Results: The fingerprints of different extracts were established and five peaks of the total 21 characteristic common peaks were identified by HPLC spectrum of standards and LCMS-IT-TOF. The strong relevance elements including macamide alkaloids represented by peaks 22-24 were verified as the potential PDE5 inhibitors. Conclusion: A sensitive and convenient screening system for the PDE5 inhibitors via liquid scintillation counting is established. Macamides as liposoluble alkaloids could be material basis components to inhibit the activity of PDE5.This paper provides certain theory basis for resource utilization and the quality control of L. meyenii.

OBJECTIVETo study clinical effects of a new internal fixation by using a cable through the bone and Kirschner with a hole in the tail, for the treatment of patellar fractures.

METHODSFrom May 2012 to July 2013, thirty-four patients with patellar fractures were treated with cable through the bone and Kirschner with a hole in the tail. All the patients had close fracture,including 12 transverse fractures and 22 comminuted fractures. There were 18 males and 16 females, ranging in age from 26 to 81 years old, with an average of (46.0 ± 3.0) years old. After open reduction, two appropriate length of Kirschner with a hole in the tail were driven into the patella as perpendicular to the fracture line or the major fragments as possible. A transverse bone tunnel was then drilled with a Kirschner at one side of the patella. Then the cable, which was successively pulled through the bone tunnel and the hole of Kirschner, was crossed in a figure-eight over the anterior of the patella, tightened and fixated by special instruments. The Kirschner was clipped off on the edge of the hole. If it was a comminuted fracture, another cable was used to fasten the patella with cerclage. Postoperative evaluation was based on Bostman.

RESULTSAll the patients were followed up, and the duration ranged from 12 to 26 months, with a mean of (16.0 ± 2.0) months. Fractures healed in all the cases without such complications as infection, loosening of Kirschner and cable loop, and skin irritation. According to the Böstman score system, 33 cases got an excellent result, and 1 good.

CONCLUSIONThe cable through the bone and Kirschner with a hole in the tail is a simple, stable and effective method for the treatment of patellar fractures, especially the transverse fractures, with earlier knee exercise and fewer complications.

Adult ; Aged ; Aged, 80 and over ; Bone Wires ; Female ; Follow-Up Studies ; Fracture Fixation, Internal ; instrumentation ; methods ; Fractures, Bone ; surgery ; Humans ; Male ; Middle Aged ; Patella ; injuries ; surgery ; Treatment Outcome ; Young Adult