Objective To investigate the Lymphocyte-specific protein-tyrosine kinase(Lck)gene ex- pression in the renal tubule epithelial cells(TECs)of lupus nephritis,and the effect of interlenkin-2(IL-2) stimulation on its expression.Methods Proximal TECs derived from 6 weeks old spontaneous systemic lupus erythematosus(SLE)BXSB mice were exposed to IL-2(100 U/ml),the expression of Lck mRNA and protein was examined by reverse transcription-polymerase chain reaction(RT-PCR)and immunoblotting respectively. The difference of Lck gene expression before and after IL-2 stimulation was investigated.The expression of Lck protein in TECs of renal tissues of BXSB mice and human with lupus nephritis was observed through im- munohistochemistry.Results The expression of Lck mRNA and protein was very low in cultured TECs of 6 weeks old BXSB mice,but increased sharply after IL-2 stimulation(P

This study was aimed to construct the targeting AATF shRNA eukaryotic expression vector and establish the stably transfected U937 cell lines. The sequence of AATF mRNA was obtained from GenBank. After excluding homology, three plasmid expression vectors coding shRNA targeting 228 ∼ 249, 303 ∼ 324 and 443 ∼ 464 of AATF gene sequence were synthesized. Two terminals of shRNA carried BamHI and HindIII restriction sites. The selected nucleotides were cloned into the plasmid pSilencer 3.1-H1 neo respectively, and the resultant recombinant plasmids were named as pSA-1, pSA-2, pSA-3. The sequences of the recombinant plasmids were identified by DNA sequencing. The recombinant plasmids were transfected into the cell line U937 by electroporation with Neon(TM) Transfection System. The transfected cells were persistently screened under G418 (500 mg/L), and isolated with a limited dilution for 8 weeks. The inhibition of AATF mRNA and protein expression was respectively detected by RT-PCR and Western blot. The results indicated that RNAi eukaryotic expression vectors targeting AATF had correct reading frame and nucleotide sequence. Real-time PCR revealed that AATF shRNA effectively silenced mRNA expression of AATF. Western blot analysis found that AATF shRNA obviously suppressed protein expression of AATF (P < 0.05). It is concluded that the shRNA eukaryotic expression vector has been successfully constructed which can inhibit the expression of AATF, and the establishment of stably transfected U937 cell lines provide a original route for exploring the mechanism of AATF in human Leukemia further.
Apoptosis Regulatory Proteins ; genetics ; Gene Expression ; Genetic Vectors ; Humans ; Plasmids ; RNA Interference ; RNA, Messenger ; RNA, Small Interfering ; genetics ; Repressor Proteins ; genetics ; Transfection ; U937 Cells

OBJECTIVETo compare the clinic outcomes of dynamic hip screw (DHS), intramedullary fixation (IF) and proximal femur locking plate (PF-LCP) in the treatment of intertrochanteric fractures in the elderly patients.

METHODSFrom July 2000 to August 2009, 165 old patients with intertrochanteric fractures were treated respectively by DHS, IF, PF-LCP. Fifty-eight patients were in DHS group including 30 males and 28 females with an average age of 71 years old; there were 30 cases of type II fracture of Jensen, 28 cases of type III fracture. Sixty-five patients were in IF group including 35 males and 30 females with an average age of 73 years old; there were 37 cases of type II fracture of Jensen, 28 cases of type III fracture. Forty-two patients were in PF-LCP group including 23 males and 19 females with an average age of 74 years old; there were 22 cases of type II fracture of Jensen,20 cases of type III fracture. The operative procedures,complications and therapeutic effects were compared among 3 groups.

RESULTSAll patients were followed up from 15 to 21 months (averaged 18.3 months). The incision length and the operation time of IF group were shorter than that of DHS and PF-LCP, but there were no significant difference between DHS group and PF-LCP group. The intraoperattive blood loss, rehabilitation and healing time of IF and PF-LCP were less or shorter than that of DHS group, but there were no significant difference between IF group and PF-LCP group. The functional recovery of IF group and PF-LCP were better than that of DHS group, there were significant difference among 3 groups. The complications of PF-LCP group was fewer than that of IF group and DHS group.

CONCLUSIONPF-LCP is the credible method for intertrochanteric fractures in the elderly patients, especially for severe comminuted fracture and osteoporosis, for it can reduce operation complications and benefit for fracture healing and hip functional recovery.

Aged ; Bone Plates ; Female ; Femur ; injuries ; physiopathology ; surgery ; Follow-Up Studies ; Fracture Fixation, Internal ; adverse effects ; instrumentation ; Hip Fractures ; physiopathology ; surgery ; Humans ; Male ; Postoperative Complications ; Treatment Outcome

AIM To investigate the preventive effects of herbal pair,Scutellariae Radix and Coptidis Rhizoma (SC),on Alzheimer's disease (AD),and its mechanism of action.METHODS Dementia mice induced by 8-week s.i.d subcutaneous injection of D-galactose (100 mg/kg),were simultaneously given respective,intragastric administration of SC crude drug at doses of 5,10,20 g/kg,or piracetam support at 0.75 g/kg,and isometrical distilled water was applied to the mice of normal control group.The mice had their learning and memory abilities checked by Morris water maze at intervals of four weeks and eight weeks since the start of the trial,and their blood and brain tissue biochemical indices measured at the end of the test.RESULTS Significantly shortened latent period in place navigation test and the time of enter into the original platform in the space exploration test were observed in the mice treated with 4-week D-galactose and SC (P ＜0.05 或 P ＜0.01).The 8-week intervention demonstrated SC capacity in the significant promotion of T-SOD activity,decreased blood MDA levels (P ＜ 0.01)and the brain AchE levels,and increased brain GSH-Px activity (P ＜ 0.01).CONCLUSION SC increases the concentration of acetylcholine in brain tissue and protects the central nervous tissue under oxidative stress,highlighting its therapeutic effect on AD.

Objective To study the effect of omeprazole on serum concentration and pharmacokinetics of oxiracetam in Chinese healthy male volunteers and established high performance liquid chromatography (HPLC) method for determination of oxiracetam in plasma.Methods This was a single center,opened,two-way crossover study.Eight healthy Chinese volunteers were screened.The concent ration of oxiracetam in plasma was detected by HPLC before and after omeprazole administ ration.Pharmacokinetics parameters were evaluated by DAS 2.1.1 pharmacokinetics programme.Statistical analysis was performed with SPSS11.5 software.Results The results showed that after oxiracetam was used alone and incombination with omeprazole,pharmacokinetic parameters of AUC0-t,AUC0-∞,MRT0-t,MRT0-∞,tmax,Cmax,t1/2,CL/Fand V/F had no significant difference.Conclusion HPLC method for determinating of oxiracetam concentration in plasma is rapid,simple,accurate and sensitive.There is no influence on the pharmaco-kinetics of oxiracetam when combined with omeprazole in humans.

Objective To explore the characteristics of gene sequence of mtDNA-COⅠof Culex pipiens pallens from differ-ent geographical regions in Shandong Province and different resistant strains from the lab and five common mosquito species, and analyze the genetic diversity of these mosquitoes.Methods Adult mosquitoes were collected from Jinan,Jining,Qingdao cities and other places in Shandong Province.The sensitive,dichlorvos-resistant,pyrethroid-resistant and propoxur-resistant strains were reared in the lab.Five species of mosquito(Cx.pipiens pallens,Cx.tritaeniorhynchus,Anopheles sinensis,Aedes al-bopictus,and Armigeres subalbatus)were collected from Jining City and identified in the lab.mtDNA-COⅠwas specifically am-plified by PCR and sequenced.The gene sequences were compared and analyzed by the biological information systems,and the phylogenetic tree was constructed.Results The amplified mtDNA-COⅠfragments of Cx.pipiens pallens from eight different cit-ies and four different resistant strains were 528 bp in length,with 67.4% A+T contents and two mutation sites.The nucleotide se-quence homology among the different geographic strains was 99.95% and the gene sequences of the four resistant strains were the same,showing a high homogeny.The amplified mtDNA-COⅠfragments of the five species of mosquitoes were 528 bp with 408 conserved sites,120 variable sites,42 parsimony informative sites and 78 singleton sites. The A+T contents were between 65.7% and 68.0%.The nucleotide sequence homology among the different mosquito species was between 86.17% and 92.05%,and the molecular identification was consistent with the traditional morphological identification. The molecular phylogenetic study showed that the different species were clustered at their own branch at the species and genus levels,while genera Armiger-es was distantly related to the others.Conclusion mtDNA-COⅠcould not serve as the molecular marker to analyze the popula-tion genetic variation and phylogenesis of Cx.pipiens pallens from different geographical regions and different resistant strains, but it has species and genus specificities,which could be used for the identification of the mosquito species and genus.

Objective To conduct an analysis of the epidemiological changes in malaria that have occurred in Shanxian county from 2002 to 2016. Methods A retrospective study was conducted and data were collected from web-based reporting system to explore the epidemiological characteristics in Shanxian county from 2002 to 2016. All individual case information was obtained from village malaria servicers organized by the local Shandong Institute of Parasitic Diseases. Results A total of 133 cases were identified as malaria in Shanxian county during this period, including 124 indigenous cases (93.2%) and 9 imported cases (6.8%). The 124 indigenous malaria cases were infected with Plasmodium vivax (P. vivax), whereas 7 of the 9 confirmed imported cases were infected with Plasmodium falciparum (P. falciparum), 1 was infected with Plasmodium ovale (P. ovale) and 1 patient was infected with P. falciparum mixed with P. vivax. The total number of malaria cases included 86 males (64.7%) and 47 females (35.3%). Age of the patients ranged from 1 to 83 years, although most (64.7%) infections occurred in the 21-to 60-year-old age group. Remarkably, 117 of the total malaria cases (98.0%) were reported from 2006 to 2011. The epidemic season was from June to October, with the peak occurring yearly from July to September. The most common occupation of the infected patients was farmer. In total, 58.1% of the cases occurred in 3 townships, namely, Fugang, Huanggang and Caozhuang. Conclusions In Shanxian county, the local malaria incidence experienced an emerge-peak-control-eliminate status. However, due to the numbers of migrant labourers returning from Africa, imported cases were continuous and presented an increasing annual trend, which became a non-negligible and a significant impediment for malaria elimination. Therefore, the need to eliminate instances of malaria reintroduction to receptive malaria-free areas should drive strategies to align with the epidemiological changes.

ObjectiveTo retrospectively analyze the clinical data of 52 patients with coronavirus disease-2019 （COVID-19） and explore the clinical efficacy of modified Sanxiaoyin on mild/moderate COVID-19 patients. MethodThe propensity score matching method was used to collect the clinical data of mild or moderate COVID-19 patients enrolled in the designated hospital of the Second Hospital of Jingzhou from December 2019 to May 2020. A total of 26 eligible patients who were treated with modified Sanxiaoyin were included in the observation group， and the 26 patients treated with conventional method were the regarded as the control. The disappearance of clinical symptoms， disappearance time of main symptoms， efficacy on traditional Chinese medicine （TCM） symptoms， hospitalization duration， laboratory test indicators， and CT imaging changes in the two groups were compared. ResultThe general data in the two groups were insignificantly different and thus they were comparable. After 7 days of treatment， the disappearance rate of fever， cough， fatigue， dry throat， anorexia， poor mental state， and poor sleep quality in the observation group was higher than that in the control group （P<0.05）， and the difference in the disappearance rate of expectoration and chest distress was insignificant. For the cases with the disappearance of symptoms， the main symptoms （fever， cough， fatigue， dry throat， anorexia， chest distress） disappeared earlier in the observation group than in the control group （P<0.01）. After 7 days of treatment， the scores of the TCM symptom scale of both groups decreased （P<0.01）， and the decrease of the observation group was larger that of the control group （P<0.01）. All patients in the two groups were cured and discharged. The average hospitalization duration in the observation group ［（12.79±2.68） d］ was shorter than that in the control group ［（15.27±3.11） d］ （P<0.01）. The effective rate in the observation group （92.31%， 24/26） was higher than that in the control group （76.92%， 20/26） . After 7 days of treatment， the lymphocyte （LYM） count increased （P<0.05）， and white blood cell （WBC） count and neutrophil （NEUT） count decreased insignificantly in the two groups. Moreover， levels of C-reactive protein （CRP）， erythrocyte sedimentation rate （ESR）， and procalcitonin （PCT） reduced in the two groups after treatment （P<0.01） and the reduction in the observation group was larger than that in the control group （P<0.01）. Through 7 days of treatment， the total effective rate on pulmonary shadow in the observation group （90.00%， 18/20） was higher than that in the control group （77.27%， 17/22） （P>0.05） and the improvement of lung shadow in the observation group was better than that in the control group （P<0.01）. ConclusionModified Sanxiaoyin can significantly alleviate fever， cough， fatigue， anorexia， chest distress， poor sleep quality， and other symptoms of patients with mild or moderate COVID-19， improve biochemical indicators， and promote the recovery of lung function. This paper provides clinical evidence for the application of modified Sanxiaoyin in the treatment of mild or moderate COVID-19.

Introduction: Advanced glycation end products (AGEs) formation is due to glycation of lipids, proteins, and nucleic acids. AGEs play a vital role in Diabetes Mellitus (DM) and cardiovascular complications and is enhanced by oxidative stress. Hordeum vulgare (Barley Grass) is a cereal plant of grass family poaceae and has been reported to exhibit antidiabetic and antioxidant characteristics and prevention of cardiovascular disease. Current experiment focused on the evaluation of antioxidant and anti-advanced glycation end products formation effect of Hordeum vulgare water extract. Methods: Free radical scavenging ability of Hordeum vulgare water extract, was evaluated using DPPH, nitric oxide, hydrogen peroxide, lipid peroxidation, FRAP and metal ion chelating. Total phenolic content of Hordeum vulgare was also analysed. In-vitro antiglycation activity was done using BSA-MGO, BSA-Glu. Phytochemical screening was performed to detect the presence of flavonoids, saponins, tannins, steroids and terpenoids. Results: The results showed Hordeum vulgare water extract ameliorated the formation of AGE and scavenge free radical from DPPH, Nitric oxide, hydrogen peroxide, Lipid peroxidation, FRAP and Metal ion chelating. Phytochemicals such as saponins, tannins, flavonoids, steroids and terpenoids were detected in Hordeum vulgare water extract. Conclusion: Hordeum vulgare water extract reveals antiglycation and free scavenging properties.

Introduction: Advanced glycation end product (AGE) is formed by the non-enzymatic glycation, leading to free radicals’ production and high oxidative stress. Accumulation of AGEs is associated with chronic diseases which are correlated with diabetes mellitus. Centella asiatica is a traditional medicinal herb used in Asia with numerous pharmacological effects such as antioxidant, wound healing, neuroprotective, and gastrointestinal treatment. This research was aim to evaluate the antioxidant, antiglycation, and phytochemical analysis of C. asiatica leaves (L.) water extract. Methods: The antioxidant activity of the plant extract was evaluated through 2,2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl (OH) radicals, nitric oxide (NO), and chelating capacity assays. Total phenolic content (TPC) was performed to determine the presence of bioactive compounds, such as saponins, which have been extensively reported in previous studies. Butylated hydroxytoluene (BHT) and ethylenediaminetetraacetic acid (EDTA) were used as positive control in the antioxidant assays, while quercetin was used in the antiglycation assays. Antiglycation effects of C. asiatica L. were evaluated through Bovine serum albumin (BSA)-MGO, BSA-glucose, and MGO scavenging assay. The data was expressed in terms of IC50. Result: The plant extract possessed significantly (p < 0.001) lower IC50 value compared to the BHT in DPPH and NO radical scavenging assay. It also showed stronger antiglycative effect with lower IC50 value compared to the quercetin in BSA-glucose and BSA-MGO assay. The TPC of C. asiatica L. was 3.20 mg/GAE/g at 100µg/ml. Conclusion: The antiglycation effects of C. asiaitca L. include the reduction of Amadori products formation, trapping α-dicarbonyl intermediates, and free radical scavenging. It is suggested that the antiglycation effects of C. asiatica L. maybe attributed by the presence of saponins and other phytochemicals.