Objective To establish a method for determining the dissolution oftorasemide sustained-release tablet in vitro and study the methodology of the determination.The consistency of the in vitro release behavior between self-prepared torasemide sustained-release tablet and original preparation were evaluated by constructed method.Methods HPLC method was applied to detect the cumulative release percentage of self-prepared torasemide sustained-release tablet and original preparation in five kinds of release media (water,0.1 mol/L hydrochloric acid solution,pH 4.5 acetate buffer,pH 6.8 phosphate buffer,and 0.1 mol/L hydrochloric acid solution turn to pH 6.8 phosphate buffer).Similarity factor (f2) was used to evaluate the similarity of release curves.Results There was a good linear relationship between the quality concentration of torasemide and peak area in the range of 1.0-12.0 μg/mL (r =0.9995).Results of precision and stability tests were good,and the RSDs for probational liquid were all lower than 2.0％.The average recovery of accuracy test was 100.04％,and RSD was 0.54％ (n =12).The homogeneity of within group of self-prepared preparation met the technical requirement,RSDs of each sampling points in six Dissolution Vessels were lower than 10.0％.The f2 factors of self-prepared torasemide sustained-release tablet and original preparation were 72,60,77,66,and 60 in five kinds of release media.Conclusion The method in the paper is suitable for the release test of torasemide,meanwhile,the self-prepared tablet shows consistent in vitro release behavior with that of the original preparation.

Objective To investigate the role of heat shock poteins(HSPs)in the pathogenesis of psoriasis.Methods Expression of HSP27,HSP70and HSP60was detected by immunohistochemical and image analysis in epidermal keratinocytes of pre-and post-treatment psoriatic lesional,and non-lesional skin in25psoriatic patients and6healthy controls.Results There was constitutive expression of HSP27and HSP70in epidermal keratinocytes of psoriatic non-lesional skin and normal controls,and the weak expression in psoriatic lesions.There was expression of HSP60in keratinocytes of psoriatic skin,but no expression of HSP60in non-lesional and normal skin.Expression of HSP27and HSP70recovered gradually in post-treatment psoriatic lesions,and no expression of HSP60was observed in lesional skin after treatment.Conclusion HSPs may play a certain role in psoriatic stress protective mechanism.

Aim To investigate the changes in the ex-pression of WNK1 in spinal cord of a rat model with bone cancer pain. Methods Female SD rats, weig-hing 170 ~200 g, were randomly divided into three groups:normal control group (group C, n=3), sham operation group ( group S, n =3 ) and bone cancer pain group ( group BCP, n =24 ) . Group C was not given any treatment, and group S was injected into the bone marrow of left tibia with 5 μl PBS solution while group BCP with 5 μl WALKER 256 mammary gland cancer cell suspension (approximately 1 × 105 cells). Mechanical paw withdrawal threshold ( MWT ) was measured at d1 before inoculation ( baseline) and d3, 6,9,10,11,12 after inoculation. Group S and C were sacrificed at d 12 while group BCP at d 3 ,6 ,9 ,12 after inoculation and spinal cord ( L4~6 ) were removed at different time points for detection of WNK1 mRNA ex-pression by qRT-PCR and WNK1 protein expression by Western blot. Results Compared with group C and S,group BCP’ s MWT started to decrease since d 3 ( P<0. 05 ) . The mRNA expression of WNK1 in spinal cord was up-regulated(P<0. 05) from d 3 showing an increasing trend over time until d 9 ( peak ) and then down-regulated to d 12 ( P >0. 05 ) while the protein expression upregulated since d6 and also showed an in-creasing trend to d 12 ( P<0. 01 ) . Conclusion The expression of WNK1 in spinal cord of a rat model with bone cancer pain increased abnormally, which may be involved in the occurrence and maintenance of a rat model with bone cancer pain.

ObjectiveTo observe the effect ofZuogui Jiangtang Jieyu Formula on the expressions of glutamate, NR2A and NR2B in hippocampus of diabetic rats with depression;To explore the mechanism of protective effect. Methods Diabetes with depression rat models were established and then were randomly divided into the model group, positive medicine group, high-, medium- and low-doseZuogui Jiangtang Jieyu Formula groups. Normal rats acted as normal group, 16 rats per group. After 28 days of administration, Open-field test was used to detect the behavior of the rats;glutamate content of hippocampus was detected by ELISA;the expressions of NR2A and NR2B in rat hippocampus were detected by immunofluorescence.Results Compared with normal group, automatic activity times of rats in model group decreased significantly (P<0.01);both glutamate content (P<0.01) and expressions of NR2A, NR2B (P<0.01) increased significantly. Compared with the model group, automatic activity times of rats in positive medicine group and high-doseZuogui Jiangtang Jieyu Formula group significantly increased (P<0.01);glutamate content dropped (P<0.01);expressions of NR2A and NR2B decreased (P<0.05).ConclusionZuogui Jiangtang Jieyu Formula can improve depressive behavior of diabetic rats with depression, which may be related to the regulation of glutamate content and expressions of NR2A and NR2B in hippocampus.

Object To study the protective effects of glycosides of cistanche (GCs) on focal cerebral ischemic rats. Methods Focal cerebral ischemia in rats was induced by 24 h occlusion of the middle cerebral artery (MCAO). The infarct area was measured by nitrobenzene thiocyanate (NBT) staining technique. The content of neurological deficits was evaluated by 0-11 scales. The activities of antioxidases and contents of MDA in ischemic brain tissue were analyzed. Results Significant decrease in infarct area and improvement in neurological deficits were observed by oral administration of GCs 125, 250 mg/kg, respectively. Significant increase in activities of SOD and GSH-Px, and significant decrease in contents of MDA in rats were also observed after 24 h MCAO. Conclusion GCs has a neuroprotective effect against focal cerebral ischemia and the effect may be related to the increase in activities of SOD and GSH-Px.

Objective To investigate the effect of intrathecal CLP257 on the bone cancer pain in rats.Methods Forty adult female Sprague-Dawley rats,weighing 180-200 g,were divided into 4 groups (n=10 each) using a random number table:sham operation group (group S),bone cancer pain group (group BCP),dimethyl sulfoxide (DMSO) group,and CLP257 group.Bone cancer pain was induced by inoculating Walker 256 mammary gland carcinoma cell suspension (about 1× 105cells) 10 μl into the medullary cavity of the left tibia.On 7th-9th days after establishment of the model,5％ DMSO 10 μl was injected intrathecally once a day in group DMSO,and 10 μg/μl CLP257 10 μl was injected intrathecally once a day in group CLP257.The mechanical paw withdrawal threshold (MWT) was measured on 1 day before establishment of the model (T0),on 1st-6th days after establishment of the model (T1-6),and at 4 h after intrathecal administration on 7th-9th days after establishment of the model (T7-9).After the last intrathecal administration,the L4-6 segments of the spinal cord were removed for determination of the expression of potassium chloride cotransporter 2 (KCC2) protein and mRNA by Western blot and fluorescent quantitative real-time polymerase chain reaction,respectively.Results Compared with group S,the MWT was significantly decreased,and the expression of KCC2 protein and mRNA was down-regulated in BCP and DMSO groups,and the MWT was significantly decreased (P＜0.05),and no significant change was found in the expression of KCC2 protein and mRNA in group CLP257 (P＞0.05).Compared with group BCP,the MWT was significantly increased,and the expression of KCC2 protein and mRNA was up-regulated in group CLP257 (P＜0.05),and no significant change was found in the parameters mentioned above in group DMSO (P＞0.05).Conclusion Intrathecal CLP257 can attenuate the bone cancer pain in rats.

Objective To evaluate the changes in the expression of isolectin B4 (IB4) and calcium gene-related peptide (CGRP) in neurons in dorsal root ganglion (DRG) of rats with neuropathic pain (NP).Methods Forty healthy male Sprague-Dawley rats,weighing 250-280 g,were divided into 2 groups (n =20 each) using a random number table method:solvent group (group S) and group NP.NP was induced by intraperitoneally injecting resiniferatoxin 210 μg/kg,and the solvent of resiniferatoxin was intraperitoneally injected in group S.The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were measured in 5 rats selected before establishing the model and at 1,3,7 and 42 days after establishing the model.Five rats were sacrificed at 1,3,7 and 42 days after establishing the model,and the L4-6 segments of the DRGs were removed to determine the expression of CGRP and IB4 in neurons using immunofluorescence.Results Compared with the baseline before establishing the model,the MWT was signilicantly decreased at 3,7 and 42 days after establishing the model,and the TWL was prolonged at 1,3,7 and 42 days after establishing the model in group NP (P＜0.05).Compared with group S,the MWT was significantly decreased at 3,7 and 42 days after establishing the model,the TWL was prolonged at 1,3,7 and 42 days after establishing the model,and the expression of IB4 and CGRP in neurons in DRGs was down-regulated at 1,3,7 and 42 days after establishing the model in group NP (P＜0.05).Conclusion Down-regulated expression of IB4 expression in neurons in DRGs may be involved in the development and maintenance of mechanical hypersensitivity to pain,and down-regulated expression of CGRP may be involved in the development and maintenance of thermal analgesia in rats with NP.

OBJECTIVE: To detect pathological variant in two patients with Chediak-Higashi syndrome (CHS) from a consanguineous family and to explore its genotype-phenotype correlation. METHODS: Clinical data was collected for this pedigree. Genomic DNA was prepared from probands' peripheral leukocytes and their relatives' fingernail. Whole exome sequencing and Sanger sequencing were carried out to detect potential variant of the LYST gene. RESULTS: The proband presented with partial oculocutaneous albinism, immunodeficiency and acidophilic inclusion body in bone marrow and blood smears. A novel homozygous nonsense variant c.8782C>T (p.Gln2928*) was identified in exon 34 of the LYST gene in the sib pair. The same variant was found to be in heterozygous status in 6 unaffected individuals from the pedigree. CONCLUSION Above result enriched the mutational spectrum of CHS and provided a basis for genetic counseling and prenatal diagnosis for this pedigree.
Chediak-Higashi Syndrome ; genetics ; Exons ; Heterozygote ; Humans ; Mutation ; Pedigree ; Sequence Analysis, DNA ; Vesicular Transport Proteins ; genetics ; Whole Exome Sequencing

OBJECTIVETo investigate clinical and imaging characteristics of delayed encephalopathy after carbon monoxide poisoning(DEACMP).

METHODSA retrospective and comparative analysis of neurological manifestations, course, periods of false-cure, prognosis and neuroimaging findings were conducted in 198 patients with DEACMP.

RESULTSOf these patients, 60.1% obtained an initial recovery within 30 d, 23.7% within 31-60 d and 14.6% over 60 d. Total clinical rate was 32.8%, and effectiveness rate 70.1%. According to imaging findings, 15.2% patients were found to have bilateral lesions of basal ganglion, and 70.0% with lesions of subcortical white matter, 12.6% with lesions of two types above mentioned and 2.1% with no lesions. The intervals between onset of illness and initial recovery were (44.6 +/- 10.1), (38.2 +/- 11.9), (61.3 +/- 17.0) d, and (7.5 +/- 2.4) d respectively. Imaging findings can demonstrate the severity of brain injury, but not necessarily parallel to the improvement of illness, for which SPECT proves more useful.

CONCLUSIONSDEACMP is involved in ischemic brain injury and a self-limited illness.