Improvement effect of crocin on cognitive impairment of Alzheimer's disease rats through DKK3 regulation of GSK-3 β/β-Catenin pathway / 中国临床药理学与治疗学

AIM: To explore the improvement effect and mechanism of crocin on cognitive impairmrnt of Alzheimer's disease (AD) rats. METHODS: The hippocampus of SD rats were injected with Aβ 25-35 to establish AD model, then rats were randomly divided into AD group, AD + low, medium, high dose of crocin groups (10, 20, 40 mg/kg) and AD + donepezil group (1 mg/kg), intraperitoneal injection treatment for 4 weeks, set sham group. Dark avoidance test and water maze test were used to evaluate the learning and memory abilities of rats, ELISA was used to detect serum Aβ content, HE staining and Tunel staining were used to determine pathological changes and neuronal apoptosis of hippocampus of rats, immunohistochemistry was used to detect the expression of Brdu, Dcx and NeuN in hippocampus of rats, and Western blot was used to detect the protein expression of Aβ, DKK3, β-catenin, p-GSK-3β/GSK-3β, Caspase-3, Bax, Bcl-2 in hippocampus of rats. RESULTS: Compared to sham group, the learning and memory abilities of AD group rats were decreased, serum Aβ content increased, the pathological change in hippocampus was serious, neuronal apoptosis was increased, the expression of Brdu, Dcx, NeuN were decreased, the protein expression of Aβ, DKK3, p-GSK-3β/GSK-3β, Caspase-3, Bax were increased, protein expression of β-catenin, Bcl-2 were decreased (P<0.01). Compared to AD group, after the treatment of doses of crocin and donepezil, the learning and memory abilities of AD rats were improved, serum Aβ content were increased, and the pathological change in hippocampus were alleviated, neuronal apoptosis were reduced, the expression of Brdu, Dcx, NeuN were decreased, the protein expression of Aβ, DKK3, p-GSK-3β/ GSK-3β, Caspase-3, Bax were decreased, the protein expression of β-catenin, Bcl-2 were increased, notely, dose-dependent effect of crocin was significant. CONCLUSION: Crocin reduced neuronal apoptosis and mediated DKK3 to regulate GSK-3β/ β-catenin pathway to improve the cognitive impairment of AD rats.

A 20-year study on microvascular autologous transplantation of submandibular gland for treatment of severe dry eye / 北京大学学报(医学版)

Severe dry eye is a refractory ophthalmologic disease. Our multidisciplinary research group treated severe dry eye by microvascular autologous transplantation of submandibular gland (SMG) during the past 20 years. The SMG, with its blood vessels and Wharton's duct, was harvested from the submandibular triangle and transferred to the temporal area. The blood vessels in the SMG were anastomosed with the temporal blood vessels using a microsurgical technique. Then, the distal end of Wharton's duct was sutured to form an opening in the upper lateral conjunctival fold. The tear was replaced by the secretion of the transplanted SMG to lubricate the ocular surface. In our study, the surgical techniques of blood vessel management were continuously modified to increase the survival rate of the transplanted SMG. A novel surgical modality of partial transplantation of SMG was established to prevent postoperative epiphora. A clinical study with the largest case number in the world was conducted and the effectiveness of transplantation of SMG for severe dry eye was fully confirmed. In order to resolve two main clinical problems including ductal obstruction resulted from low secretion rate during the latent period, and epiphora due to over secretion of the transplanted SMG in the later term of transplantation, the regulation of the secretion mechanism of the normal and transplanted SMG were investigated. New opinions on mechanisms of saliva secretion were provided. Based on the priniciple of translational medicine, the results of related basic research were applied in the clinic. The clinical guidelines for secretion regulation of transplanted SMG were established. A concept of chronic obstructive sialadenitis of transplanted SMG was provided and its diagnostic criteria, diagnostic technique of sialography, and therapeutic regimen were established. As a result, the surgical success rate was obviously elevated, the surgical complications were decreased, and life quality of the patients was greatly improved.

Role of microRNA-126-5p in myocardial injury induced by doxorubicin / 中国病理生理杂志

AIM:To observe the expression of microRNA-126-5p during myocardial injury and its role in myo-cardial cell injury induced by adriamycin(also called doxorubicin, DOX).METHODS: The BALB/c mouse model of DOX-induced acute and chronic myocardial injury was established via intraperitoneal injection of DOX.HE staining was applied to observe the morphological changes of myocardial tissues.Lactate dehydrogenase(LDH)in serum was detected and PowerLab system was used to detect the influence of DOX on the changes of ±dp/dtmax.The expression of microRNA-126-5p in injured myocardial tissues and the H 9c2 cells exposed to DOX was detected by real-time PCR.Gain-and loss-of-function experiments were conducted to detect the role of microRNA-126-5p in H9c2 cells treated with DOX on LDH release and caspase-3 activation.RESULTS:In acute and chronic DOX myocardial damage models in mice,HE staining showed disarranged myocardial fibers, dissolved myofibril and inflammatory cell infiltration.Higher serum LDH level and lower ±dp/dtmaxin DOX-treated mice than those in normal mice were found.Compared with the normal mice, the expression level of microRNA-126-5p was significant increased in the myocardium with DOX-induced injury.Similarly,the expression level of microRNA-126-5p was significant increased in the H9c2 cells treated with DOX.In addition, over-expression of microRNA-126-5p decreased cell viability and promoted apoptosis,while microRNA-126-5p ablation promoted the viability and inhibited the apoptosis of H9c2 cells.CONCLUSION:The microRNA-126-5p expression is up-regulated in myocar-dial injury induced by DOX,and microRNA-126-5p inhibits cell viability and promotes apoptosis induced by DOX.

Changes of Peripheral Blood Marrow-Derived Suppressor Cell Level after Chemotherapy Induction Remission by VDLP Regimen and Their Relationship with Immune System in B-ALL Children / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the changes of peripheral blood marrow-derived suppressor cell level after chemotherapy induction remission by regimen consisting of vincristine, daunorubicin, L-asparaginase and prednisone (VDLP) and to analyze their relationship with immume system in B-ALL children.</p><p><b>METHODS</b>Thirty B-ALL children after induction remission by VDLP regimen from August 2015 to August 2016 were selected as B-ALL group and 30 normal healthy children were selected as control group. The peripheral blood in 2 groups was collected and detected by flow cytometry, then the ratios of CD30cells and CD33HLA-DRmarrow-derived suppressor cells, CD14CD33HLA-DRmarrow-derived suppressor cells and CD15CD33HLA-DRmarrow-derived suppressor cells were calculated, and their changes after induction remission by VDLP regimen and the relationship with immune system were analyzed.</p><p><b>RESULTS</b>After treatment the ratio of CD33cells in peripheral blood of B-ALL group and control group was not significantly different (P> 0.05), moreover, the ratio of CD33cells in B-ALL group was significantly higher than that before treatment (P<0.05), while the ratios of CD33HLA-DRmarrow-derived suppressor cells, CD14CD33HLA-DRmarrow-derived suppressor cells and CD15CD33HLA-DRmarrow-derived suppressor cells in B-ALL group were significantly lower than those in control group (all P<0.05), but the ratios of these cells in B-ALL group were higher than those before treatment, and yet there was no statistical significance (P>0.05).</p><p><b>CONCLUSION</b>The ratios of marrow-derived suppressor cells in peripheral blood of B-ALL children in complete remission after treatment with VDLP regimen are higher than those before treatment, but are significantly lower than normal value, which may be related with non-complese recovery of immune system in B-ALL children after treatment.</p>

Changes of pharmacokinetics of 6,7-dimethoxycoumarin in a rat model of alpha-naphthylisothiocyanate-induced experimental hepatic injury after Yinchenhao Decoction () treatment / 中国结合医学杂志

<p><b>OBJECTIVE</b>To study the changes of pharmacokinetics of 6,7-dimethoxycoumarin in a rat model of alpha-naphthylisothiocyanate (ANIT)-induced experimental hepatic injury after oral administration of Yinchenhao Decoction (, YCHD) using an ultra pressure liquid chromatography (UPLC) method.</p><p><b>METHODS</b>Rats were divided into a normal group and a model group, after modeled by 4% ANIT (75 mg/kg) for 48 h, they were orally administrated with YCHD extract at the dose of 0.324 g/kg, and then blood was collected from their orbital sinus after different intervals. Changes in liver function were monitored by the levels of liver enzymes [alanine aminotransferase (ALT), aspartate aminotransferase (AST)] and bilirubins [total bilirubin (TBIL), direct bilirubin (DBIL)], the concentration of 6,7-dimethoxycoumarin in plasma were measured by UPLC, and the pharmaceutical parameters were calculated with DAS2.1.1 software.</p><p><b>RESULTS</b>The concentration-time curve of both normal and modeled rats after oral administration of YCHD was obtained. Their time to maximum plasma concentration (t(max)) were both 0.25 h, the maximum concentration (C(max)) were 4.533 μg/mL and 6.885 μg/mL, and their area under concentration-time curve (AUC)(0→24h) were 16.272 and 32.981, respectively. There was a 51.88% and 100.46% increase in C(max) and AUC(0-t) (P<0.05), but there showed a 45.52% and 92.93% reduction in clearance of drug and volum of distribution (P<0.05), respectively.</p><p><b>CONCLUSIONS</b>Hepatic injury could significantly influence the pharmacokinetics of 6,7-dimethoxycoumarin after oral administration of YCHD, the absorption and distribution process was accelerated in liver injured rats, but the metabolism and elimination process was slowed. And this may lead to a significant accumulation of 6,7-dimethoxycoumarin in the body.</p>

The effect of pulsed electromagnetic fields on the expression of transforming growth factor beta-1 after sciatic nerve injury / 中华物理医学与康复杂志

Objective To observe any effects of pulsed electromagnetic fields (PEMFs) on the expression of transforming growth factor beta-1 (TGF-β1) after sciatic nerve injury and to investigate the possible mechanism of any regeneration of the injured sciatic nerve.MethodsForty-eight SD rats were randomly divided into a PEMF treatment group,a model group and a normal control group with 16 rats in each group.The three groups were then sub-divided into 1 day,3 day,7 day and 14 day subgroups.The rats of the model and treatment groups were clamped to produce a sciatic nerve injury model.The treatment sub groups were exposed to a 9 mT PEMF at 14 Hz for 2 hours once daily for 1,3,7 and 14 days,respectively.The model group was given sham exposure and the normal control group was reared conventionally and not given any special treatment.The histological changes in the rats' sciatic nerves were observed under a light microscope after hematoxylin-eosin staining.Expression of TGF-β1 was detected by immunohistochemical methods.ResultsAfter 7 and 14 days of treatment,Wallerian degeneration of the sciatic nerve in the treatment group was more obvious than in the model group.The expression of TGF-β1 increased during the treatment process and reached a maximum at the 14th day after nerve injury.The expression of TGF-β1 had increased significantly in the model and treatment groups compared with the control group at all observation time points.At the 3rd,7th and 14th day after the operation,the expression of TGF-β1 in the treatment group was significantly higher than that in the model group. Conclusion PEMFs can accelerate Wallerian degeneration of peripheral nerves and can up-regulate the expression of TGF-β1 after sciatic nerve injury,at least in rats.

Establishment and application of nested real-time quantitative polymerase chain reaction assay for detection of hepatitis B virus covalently closed circular DNA / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To establish a nested real-time quantitative polymerase chain reaction (PCR) assay for detection of hepatitis B virus covalently closed circular DNA in PBMC( peripheral blood monocyte) and MMNC (marrow monocyte).</p><p><b>METHODS</b>Based on the structural differences between HBVcccDNA and HBV rcDNA, two pairs of specific primers spanned the gap of the positive and negative chains and a specific TaqMan probe situated downstream were designed. To remove rcDNA, cccDNA was processed by Mung Bean Nuclease,and then amplified by nested real-time quantitative PCR using a pair of outer primers and a pair of inner primers. According to the standard preparation, cccDNA levels of specimen were calculated.</p><p><b>RESULTS</b>We have established a nested real-time fluorescent quantitative PCR method for HBV cccDNA successfully, and the linear range is from 5.0 x 10(2) to 3. 9 x 10(7) copies per milliliter. Of the 25 PBMC samples and 7 MMNC samples of the chronic hepatitis B or liver cirrhosis patients, 3 MMNC samples and 9 PBMC samples were HBV cccDNA positive, while all of the 21 healthy donator blood PBMC samples were negative.</p><p><b>CONCLUSIONS</b>The nested real-time fluorescent quantitative PCR method may be applied to detect HBVcccDNA level in PBMC and MMNC. HBVcccDNA can be detected in PBMC and MMNC.</p>

The detection and clinical significance of hepatic fibrosis index / 中华实验和临床病毒学杂志

<p><b>OBJECTIVE</b>To study the relationship between levels of serum HA, LN, IV-C, PC III of chronic hepatitis and indexes of hepatic fibrosis.</p><p><b>METHODS</b>The levels of serum HA, LN, IV-C and PC III of chronic hepatitis of 124 cases and health 18 cases were measured by radio immunoassay, combined with clinical characteristics and 33 cases pathologic slice etc. The diagnostic of the indexes of serum was analyzed with statistics.</p><p><b>RESULTS</b>HA and IV-C are parallel in chronic hepatitis periods. LN and PC III are concert in the same pathologic periods. In G4 period PC III is nearly closed with comparative group. The value of HA, LN, NV-C and PC III in the chronic hepatitis group was significantly higher than that in the normal comparative group. Conclusion The levels of serum HA LN IV-C and PC III are in concert with the degree of hepatic fibrosis, and these indexes are valuable for chronic hepatitis diagnoses combined with the clinic. LN and PC III are coincidence with hepatic fibrosis degree before G4 period.</p>

The effect of pulsed ultrasound on medial collateral ligament repair / 中华物理医学与康复杂志

Objective To investigate the effect of pulsed ultrasound therapy (US) on medial collateral liga-ment healing in rats and it's mechanism.MethodsEighteen 3-month-old male Sprague-Dawley rats with transected medial collateral ligaments (MCLs) were studied. They were randomly divided into a control group, a 0.5 W/cm~2 group and a 1. 0 W/cm~2 group.The control group was not given any treatment.The 0. 5 W/cm~2 group and 1.0 W/cm~2 group were given 10 minutes of pulsed US (duty cycle: on/off = 3 ms/1 ms) daily for 8 days at either 0.5 or 1.0 W/cm~2 intensity. All the rats were sacrificed on the 9th day. After macroscopic examination, their MCLs were harvested and studied using haematoxylin-eosin staining, Van Gieson's staining and immunohistochemical tech-niques in order to detect transforming growth factor beta-1(TGF-β1) and any histological or histochemical changes.ResultsMacroscopically, the lacerated MCLs had healed with scar tissue formation. Scarring appeared to be greater in the 0.5 W/cm~2 and 1.0 W/cm~2 groups than in the control group. Inflamed cells appeared to be more numerous in the treated groups than in the controls. There were significant differences in collagen fiber extent among all three groups. In the 1.0 W/cm~2 group, the average level of TGF-β1 was significantly up-regulated, and TGF-β1 expres-sion was higher than in the other two groups.ConclusionsPulsed US can improve ligament healing in the short term, however whether long-term treatment with US can yield further improvement is unknown. Pulsed US can in-crease the level of TGF-β1, which will be higher with higher US dosage. Pulsed US may enhance injored ligament re-pair by up-regulating TGF-β1.Objective To investigate the effect of pulsed ultrasound therapy (US) on medial collateral liga-ment healing in rats and it's mechanism.MethodsEighteen 3-month-old male Sprague-Dawley rats with transected medial collateral ligaments (MCLs) were studied. They were randomly divided into a control group, a 0.5 W/cm~2 group and a 1. 0 W/cm~2 group.The control group was not given any treatment.The 0. 5 W/cm~2 group and 1.0 W/cm~2 group were given 10 minutes of pulsed US (duty cycle: on/off = 3 ms/1 ms) daily for 8 days at either 0.5 or 1.0 W/cm~2 intensity. All the rats were sacrificed on the 9th day. After macroscopic examination, their MCLs were harvested and studied using haematoxylin-eosin staining, Van Gieson's staining and immunohistochemical tech-niques in order to detect transforming growth factor beta-1(TGF-β1) and any histological or histochemical changes.ResultsMacroscopically, the lacerated MCLs had healed with scar tissue formation. Scarring appeared to be greater in the 0.5 W/cm~2 and 1.0 W/cm~2 groups than in the control group. Inflamed cells appeared to be more numerous in the treated groups than in the controls. There were significant differences in collagen fiber extent among all three groups. In the 1.0 W/cm~2 group, the average level of TGF-β1 was significantly up-regulated, and TGF-β1 expres-sion was higher than in the other two groups.ConclusionsPulsed US can improve ligament healing in the short term, however whether long-term treatment with US can yield further improvement is unknown. Pulsed US can in-crease the level of TGF-β1, which will be higher with higher US dosage. Pulsed US may enhance injored ligament re-pair by up-regulating TGF-β1.

Advance in Rehabilitation of Diabetic Peripheral Neuropathy (review) / 中国康复理论与实践

@#Diabetic peripheral neuropathy (DPN) is the most common complaint of diabetic patients, affecting the quality of life of patients seriously. The effect of DPN treated with traditional medicine therapy is poor. The rehabilitation of DPN develops fast recently. The authors overview the current trend of the rehabilitation treatments of DPN in the article including medicine therapy, physical therapy, exercise therapy, traditional Chinese medicine, hyperbaric oxygen therapy, orthopaedic devices and so on.

Clinical and Gene Diagnosis on Menkes Disease in Two Families / 实用儿科临床杂志

Objective To detect the mutation of ATP7A gene in 2 families with Menkes disease.Met-hods Genomic DNA of 6 members from 2 families were extracted with salt fractionation.The encoding exons and 2 sides flanking intron of ATP7A gene were amplified from genomic DNA of the probands and their parents by PCR and directly sequence.Light microscope was used to test the hair of probands and normal healthy children.Results Proband 1 had a deletion mutation of c.3 045del T in exon 14 of ATP7A gene and resulted in a stop codon just several nucleotides behind the deletion site.His mother was a heterozygote of the mutation and had normal phenotype.Proband 2 had a nonsense mutation of c.2 956 in exon 14 of ATP7A gene and resulted in a stop of amino acid synthesis.His mother was not a heterozygote of the mutation.Genetype and phenotype in fathers of the 2 probands were normal.Hair of the probands in light microscope were tenuity,midheaven,the color of hair also turned to light.Conclusions The c.3 045del T mutation of ATP7A gene cause the phenotype of Menkes disease in proband 1.His mother is a heterozygote of the disease without symptoms and he is of familial inheritance.The c.2 956 nonsense mutation of ATP7A gene cause the phenotype of Menkes disease in proband 2.His mother is not a heterozygote of the disease and he is not of familial inheritance.

Follow up of neonata hypoxic ischemic encephalopathy and ventricular dilatation / 实用儿科临床杂志

Objective To investigate the prognostic sequelae in neontes with hypoxic-ischemic encephalopathy (HIE) and ven-tricnlar dilatation.Methods Seventy-six full term newborns infants with HIE were followed up at the age from 3 to 19 months after therapy. Twenty-five infants among them were followed up by telephone in the epidemic period of SARS.Results Among 76 infants of 88 newborn infants with HIE(84.6%), 73 infants were normal (96.1% ). 1 infant had cerebral palsy (1.3%), 2 infants died (2.6 %).Among 39 cases with mild HIE, none of them had cerebral sequelae; among moderate HIE. 1 infant had cerebral palsy (2.9%) 1 infant died (2. 9 %), interlenkin-4 among severe HIE 50 % died (P00.5 The poor outcome of HIE in those infants were related to intrauterine growth retardation,severe birth asphyxia;and inadequate treatment.Cranial ultra-sonography of 49 infants were done on follow-up,and 12 of them (24.5 % ) had ventricular dilatations, which appeared after birth with 6 infants. Others occurred on follow-up with 1 infant had cerobral palsy,all ventricular dilatations recovered to normal at 12- 19 months except the cerebral palsy.Conclusions The poor outcome of HIE depends on the infants with intranterine growth relarda-tion,severe birth asphyxia and inadequate treatment.The prognosis of transient ventrealar ddatation are good except cerebral palsy.J Appl Clin pediatr,2004,19(12) : 1045- 1047

Clinical analysis of stereotactic radiotherapy combined with Gemzar for treatment of advanced pancreatic cancer / 中国现代普通外科进展

Objective: To observe the efficacy of stereotactic radiotherapy combined with gemzar for treatment of unresectable pancreatic cancer.Methods: Gemzar at dose of 1000mg/m 2,28 days per cycle,drug was given at 1,8,15 days,for two cycles.After 24 hours all patients were treated with stereotactic radiotherapy.Per fraction was 5～7G Y,three times per week.The total dose was 40～48G Y.Results: 13 patients were followed up for one year,symptoms were reduced and median survival was 12 months.Conclusion: Stereotactic radiotherapy combined with gemzar has important valves for unresectabal pancreatic cancer.