Disinfectants ; analysis ; Disinfection ; Drinking Water ; analysis ; Environmental Monitoring ; methods

Background Recently,there were many studies on corneal innervations during mammalian development.However,there were fewer studies on discussing corneal innervations before and after mouse eye openings.Objective The present study was to investigate the change in the regulation of corneal nerve fiber length and density before and after mouse eye openings to offer a basis for clinical research in human.Methods Thirty SPF C57BL/6 mice were divided into postnatal 1 day(P1 d),P7 d,P13 d(1 day before eye opening),P14 d(eye halfopened),P17 d(1 day after eye opening)and P23 d(7 day after eye opening)groups,with 5 mice and 10 eyes for each group.Entire corneal stretches were prepared and immunostaining with an anti-neuron-specific β-Ⅲ tubulin antibody was performed to label the corneal nerve fibers.Confocal microscopic pictures from the corneal dorsal-nasal region (DN),dorsal-temporal(DT),ventral-nasal region(VN)and ventral-temporal(VT)were taken using Delta Vision Core.From these pictures,the mouse corneal area,total length and density of nerve fibers in the 4 regions were calculated.The use of the animals complied with Statement of ARVO.Results Corneal areas of P1 d,P7d,P13 d,P14 d,P17 d and P23 d mice were(0.404±0.007),(1.362±0.154),(1.573±0.080),(1.603±0.046),(1.847±0.052),(2.445±0.798)mm2,respectively ; the total lengths of nerve fibers were(3.718±1.044),(19.065±3.350),(23.687±0.907),(27.309±2.477),(31.989±3.976),(41.214±1.573)mm,respectively ; the densities of nerve fibers were(9.592±1.138),(14.506±1.908),(15.088±1.241),(16.772±1.897),(16.821±2.102),(17.660±1.216)mm/mm2,respectively,all showing significant increases with age(F =22.906,P =0.000 ; F =0.424,P =0.000 ; F =2.375,P=0.000).A positive correlation of the increasing corneal areas and increasing lengths of nerve fibers was found(r=0.983,P＜0.01).Nerve fiber densities in the four corneal regions significantly increased with age(DN region:F =0.159,P =0.000 ; DT region:F =2.1 72,P =0.001 ; VN region:F =1.998,P =0.000 ; VT region:F=2.352,P=0.000).From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference(t =0.589,P =0.572); and the corneal nerve fiber densities in the DT region,VN region and VT region decreased by 4.6％,5.5％ and 0.1％,respectively,without significant difference from P14 d to P17 d(t=0.549,P=0.596;t=0.701,P=0.501 ;t=-0.100,P=0.919).Conclusions The development of nerve fibers in the whole cornea or the four corneal regions is influenced by eye opening in mouse to various extents.From P13 d to P14 d,the corneal nerve fiber densities in the DN region decreased by 6.0％ without significant difference.From P14 d to P17 d,the corneal nerve fiber densities in the DT region,VN region and VT region decrease by 4.6％,5.5％ and 0.1％,respectively,without significant difference.Afterwards,the growth of nerve fibers increased in pace and the growth rate is recovered.

OBJECTIVETo analyze the mutation of the KAL1 gene in male patients with idiopathic hypogonadotropic hypogonadism (IHH).

METHODSWe analyzed the exon mutation of the KAL1 gene in 30 IHH patients using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) combined with the PCR product direct sequencing technique.

RESULTSThree cases of the KAL1 gene mutation were found among the total number of patients, including 1 case of nonsense mutation (c. 1270C > T,p. R424X), and 2 cases of frameshift mutation, (c. 279_280delAG,p. G94fs) and (c. 1886_1887delTT,p. L629fs).

CONCLUSIONOf the 3 cases of the KAL1 gene mutation we detected, 2 are new and 1 already reported in the literature. The results of our study have provided valuable information on the molecular genetics of the IHH syndrome.

Adolescent ; Adult ; Base Sequence ; Child ; DNA Mutational Analysis ; Exons ; Extracellular Matrix Proteins ; genetics ; Humans ; Hypogonadism ; genetics ; Kallmann Syndrome ; genetics ; Male ; Mutation ; Nerve Tissue Proteins ; genetics ; Polymorphism, Single-Stranded Conformational ; Young Adult

Objective Prospective comparison was done on concurrent chemo-radiotherapy and se- quential chemo-radiotherapy for unresectable stageⅢnon-small cell lung cancer(NSCLC) and to evaluate three different regimens of concurrent chemo-radiotherapy.Methods Ninety-six such patients were ran- domized into four groups:1.sequential chemo-radiotherapy group received two cycles of induction chemother- apy with 40 mg/m~2 of cisplatin on D 1-3,29-31 and 100 mg/m~2 of etoposide on D 1-3,29-31 before conven- tional radiotherapy,2.concurrent chemo-radiotherapy group 1 received 100 mg/m~2 etoposide on D 1-3 and DDP 40 mg/m~2 on D 1-3,D 29-31,iv.drip,3.concurrent chemo-radiotherapy group 2 received concurrent chemotherapy with 40 mg/m~2 of paclitaxel every Monday during conventional radiotherapy,4.concurrent chemo-radiotherapy group 3 received concurrent chemotherapy with 40 mg/m~2 of paclitaxel every Monday during three-dimensional conformal radiotherapy.All patients were irradiated with 2.0 Gy/fraction,5 frac- tions/week,to a total dose of 60-64 Gy.They all received two cycles of consolidation themotherapy with 40 mg/m~2 of cisplatin on D 1-3 and 100 mg/m~2 of etoposide on D 1-3.Results The overa/1 response rate was 67%,71%,71% and 79% for sequential ehemo-radiotherapy group,concurrent chemo-radiotherapy group 1,2 and 3,respectively.There was a significant difference between the concurrent chemo-radiotherapy and sequential chemo-radiotherapy(P＜0.05).The 1-,3-and 5-year overall survival rate(OS) was 54%,8% and 4%;71%,17% and 8%;79%,17% and 8%;83%,46% and 13%,respectively for the four groups. The difference among all these groups(P=0.017) was significant.It was also significant between the con- current chemo-radiotherapy group 1 and 3 (P=0.046).The difference of distant metastasis rate among all the groups was statistically insignificant (P＞0.05) also was the difference of toxicity (P＞0.05),but the severe toxicity of concurrent chemo-radiotherapy groups 1 and 2 were higher than the sequential chemo-radio- therapy group and concurrent chemo-radiotherapy group 3.Conclusions Better locoregional progression- free survival and overall survival of unresectable stageⅢnon-small cell lung cancer could be achieved by concurrent chemo-radiotherapy as compared with sequential chemo-radiotherapy though at the expense of in- crease in toxicity.With the combination of concurrent chemo-radiotherapy and conforrnal radiotherapy,the o- verall survival rate could be much improved with miider toxicity.

OBJECTIVETo investigate the effect of ionization on the expression of hypoxia-inducible factor-1alpha and vascular endothelial growth factor (VEGF) in human hepatocellular carcinoma HepG2 cells under anoxic condition, and search for an effective method for improving the radiosensitivity of the tumor cells.

METHODSHepG2 cells were divided into 4 groups, namely the control group, hypoxia group, ionization radiation group, and hypoxia and radiation group, with corresponding treatments. The cell apoptosis was detected by fluorescence microscope, and the cell viability analyzed by MTT assay. The expressions of HIF-1alpha and VEGF mRNAs were detected by RT-PCR.

RESULTSA few apoptotic cells were found in hypoxia group, but significant apoptosis occurred in the radiation group; fewer apoptotic cells were observed in the hypoxia and radiation group than in the hypoxia group. The viable cell fraction increased in the order of the control group>hypoxia group> hypoxia plus radiation group> radiation group (P<0.05), and the expression of HIF-1alpha mRNA increased in the order of hypoxia plus radiation group>hypoxia group>radiation group (P<0.05), and no significant difference was found in the radiation group and control group. The expression of VEGF mRNA increased in the order of hypoxia plus radiation group> hypoxia group>radiation group>control group (P<0.05).

CONCLUSIONThe expression of HIF-1alpha may protect the hepatocellular carcinoma cells from damages by radiation in hypoxic condition, and HIF-1alpha decreases the radiosensitivity of the cells possibly by inducing VEGF expression.

Apoptosis ; Carcinoma, Hepatocellular ; metabolism ; radiotherapy ; Cell Hypoxia ; Cell Survival ; Hep G2 Cells ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; metabolism ; radiation effects ; Radiation, Ionizing ; Vascular Endothelial Growth Factor A ; metabolism ; radiation effects

OBJECTIVETo analysis the changes of two chemical constituents, namely 2, 3-dihydro-3, 5- dihydroxy-6-methyl-4H-pyran-4-one (DDMP) and 5-hydryoxymethyl-furfural (5-HMF) produced in Radix Polygoni Multiflori after processing, with processing time, and to determine the contents of 5-HMF in samples of Radix Polygoni Multiflori and Radix Polygoni Multiflori preparata.

METHODAn HPLC method was applied with a Zobax SB-C18 (3.9 mm x 150 mm, 5 microm) column by a elution using methanol-water (10: 90) as the mobile phase. The detection was set at UV 280 nm.

RESULTThe contents of DDMP were increasing with the processing time until 24 hour, followed by a decrease until 60 hour process. The contents of 5-HMF were increasing gradually throughout the 60 hour steaming process. The contents of 5-HMF in 11 samples of Radix Polygoni Multiflori preparata were from 0.013% to 0.101%, and only one in 4 samples of Radix Polygoni Multiflori containing trace amount of 5-HMF.

CONCLUSIONThe chemical components in Radix Polygoni Multiflori were changed during the processing procedures. Therefore, the processing of Radix Polygoni Multiflori should be controlled and standardized.

Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; chemistry ; Polygonaceae ; chemistry ; Reproducibility of Results

The main factors which affected the isolation, purification and cultivation of Pinellia cordata protoplasts from leaves were studied. The results indicated that the optimum enzyme solution for P. cordata leaves was 13% CPW + 1.0% Cellulose +0.1% Pectolase, at pH 6.0, temperature (25-28 degrees C ) for 4 h. The sucrose density gradient centrifugation was adopted to purificate the protoplasts collected, when 25% sucrose was used as mediator, centrifugating at 500 rpm for 10 min. When the protoplasts were shallow liquid and liquid-solid double layer cultured on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA + 13% mannitol at the density of 2.5 x 104 protoplasts/mL, or fed and nursed cultured at the density of 100-500 protoplasts/mL, cell division could be observed for 3 days; granular calli appeared for 30 days. Calli was proliferated on the medium of MS + 0.5 mg x L(-1) 6-BA + 0.25 mg x L(-1) NAA solidified by 0.55% agar, and differentiated and regenerated after 5-6 months. Plant generation of P. cordata is successfully established.
Cell Separation ; methods ; Culture Media ; Pinellia ; physiology ; Protoplasts ; physiology ; Regeneration

OBJECTIVETo study the clinicopathological features of patients with urothelial carcinoma of the urinary bladder (UCB), and analyze the association of clinicopathological characteristics with tumor recurrence and progression.

METHODSAltogether 658 UCB cases in Fudan University Shanghai Cancer Center were collected from January 2006 to December 2010. The histopathologic materials and the clinical records were reviewed. Univariate and multivariate analyses were preformed to detect the association.

RESULTSThe mean age of the patients was 61.97 +/- 12.97 years (range, 20-90 years). Male to female ratio was about 5:1. A total of 517 cases (78.6%) were superficial at the time of diagnosis (stage Ta/T1). The mean follow-up period was 22.36 +/- 24.92 months. Twenty-five patients lacking follow-up information were excluded in calculating recurrence and progression rates, the recurrence rate was about 37.0% (234/633), and progression rate about 6.2% (39/633). Three variables (grade, tumor growth pattern, and pathological stage) were found to be significant risk factors for tumor progression in univariate and multivariate analyses (P < 0.05).

CONCLUSIONSMost of the newly diagnosed UCB cases may be superficial diseases. Grade, tumor growth pattern, and pathological stage are associated with tumor progression of UCB.

Adult ; Aged ; Disease Progression ; Female ; Humans ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; pathology ; Prognosis ; Urinary Bladder Neoplasms ; pathology

BACKGROUNDThe auditory brainstem implants (ABIs) have been used to treat deafness for patients with neurofibromatosis Type 2 and nontumor patients. The lack of an appropriate animal model has limited the study of improving hearing rehabilitation by the device. This study aimed to establish an animal model of ABI in adult rhesus macaque monkey (Macaca mulatta).

METHODSSix adult rhesus macaque monkeys (M. mulatta) were included. Under general anesthesia, a multichannel ABI was implanted into the lateral recess of the fourth ventricle through the modified suboccipital-retrosigmoid (RS) approach. The electrical auditory brainstem response (EABR) waves were tested to ensure the optimal implant site. After the operation, the EABR and computed tomography (CT) were used to test and verify the effectiveness via electrophysiology and anatomy, respectively. The subjects underwent behavioral observation for 6 months, and the postoperative EABR was tested every two weeks from the 1 st month after implant surgery.

RESULTThe implant surgery lasted an average of 5.2 h, and no monkey died or sacrificed. The averaged latencies of peaks I, II and IV were 1.27, 2.34 and 3.98 ms, respectively in the ABR. One-peak EABR wave was elicited in the operation, and one- or two-peak waves were elicited during the postoperative period. The EABR wave latencies appeared to be constant under different stimulus intensities; however, the amplitudes increased as the stimulus increased within a certain scope.

CONCLUSIONSIt is feasible and safe to implant ABIs in rhesus macaque monkeys (M. mulatta) through a modified suboccipital RS approach, and EABR and CT are valid tools for animal model establishment. In addition, this model should be an appropriate animal model for the electrophysiological and behavioral study of rhesus macaque monkey with ABI.

Animals ; Auditory Brain Stem Implants ; Deafness ; surgery ; Evoked Potentials, Auditory, Brain Stem ; physiology ; Female ; Macaca mulatta ; Male

Kallmann syndrome (KS) is a clinically and genetically heterogeneous disorder that occurs in either an inherited or a sporadic manner. KS results from failed embryonic migration of GnRH-1 neurons from the nasal placode to the hypothalamus, due to the abnormal development of olfactory nerves and bulbs. Hypogonadotropic hypogonadism is related to GnRH deficiency, and anosmia is associated with the absence or hypoplasia of olfactory bulbs and tracts. KS patients can also present some non-reproductive or non-olfactory anomalies in addition to the above typical symptoms. For the high complexity of the molecular genetic mechanism of KS, to date, only 6 KS-related genes have been identified. The KAL1 gene is responsible for the X chromosome-linked recessive form of KS, while the fibroblast growth factor receptor 1 (FGFR1/KAL2) and fibroblast growth factor 8 (FGF8/KAL6) genes are related to the autosomal dominant form of the disease. However, the mutations in these 6 genes account for only about 25 - 30% of all KS cases, which suggests that other pathogenic genes involved in KS remain to be discovered. This article presents an overview on the studies of the pathogenic genes, clinical diagnosis and treatment of KS.
Extracellular Matrix Proteins ; genetics ; Humans ; Kallmann Syndrome ; genetics ; Mutation ; Nerve Tissue Proteins ; genetics ; Receptor, Fibroblast Growth Factor, Type 1 ; genetics