Nucleic acid sequencing was proceeded to examine the distribution of iNOS gene polymorphisms in type 2 diabetes in Singapore. There were significant differences in allele distribution between different ethnic groups, but the polymorphisms failed to display any association with DN.

Abstract: Objective To investigate the nutritional status of patients with pulmonary tuberculosis and its effects on conventional anti-tuberculosis treatment, so as to provide a basis for improving the efficacy of conventional treatment of pulmonary tuberculosis. Methods The relevant data of 168 patients with pulmonary tuberculosis admitted to Suining Central Hospital from April 2020 to April 2022 were retrospectively analyzed. Nutritional status of the patients before treatment was investigated using the Mini Nutritional Assessment (MNA) score, and the influencing factors of nutritional status before treatment were analyzed. Therapeutic effects of anti-tuberculosis drugs in the non-nutritional risk group and the nutritional risk group were comparatively analyzed. Results Among the 168 patients, 64 were assessed as having good nutritional status before treatment, 59 had the risk of malnutrition and 45 were malnourished according to the MNA score. Univariate analysis and linear regression analysis showed that age, underlying diseases, and clinical symptoms were factors affecting the MNA score before treatment (t=3.173, 3.718, 2.018, P all<0.05); whereas gender and education level were not factors affecting MNA score before treatment (t=0.065, 0.059, P all>0.05). According to the MNA score before treatment, the patients were dividedinto a non-nutritional risk group (MNA score > 23.5) and a nutritional risk group (MNA score ≤23.5). The negative conversion rate of sputum bacteria, effective rate of focal absorption in the non-nutritional risk group were 92.19% (59/64)and90.63% (58/64) , respectively, which were significantly higher than corresponding 79.85% (82/104)and76.92% (80/104) in the nutritional risk group. The drug resistance rate, adverse reaction rate, and average treatment cost of the no nutritional risk group and nutritional risk group were 7.81% (5/64) and 21.15% (12/104), 15.63% (10/64) and 31.73% (33/104), (0.62±0.13) million yuan and (0.89±0.26) million yuan, respectively, with significant differences (χ2=5.228, 5.071, 7.685, 5.396, 7.728, P all<0.05). Conclusions Patients with pulmonary tuberculosis exhibit poor nutritional status before treatment. The patients’nutritional status is easily affected by age, underlying diseases, and clinical symptoms, thereby affecting the effect of anti-tuberculosis treatment. Therefore, early nutritional intervention for tuberculosis patients should be recommended in order to prevent malnutrition and enhance the effectiveness of anti-tuberculosis treatment.

Objective To examine the association of intron 4 polymorphism in endothelial nitric oxide synthase ( eNOS) gene with diabetic nephropathy and its distribution in different ethnic groups of Singapore. Methods 258 patients with diabetic duration longer than 10 years (150 Chinese, 71 Malay and 37 Indian) were selected from both inpatients and outpatients of National University Hospital. Polymorphism was identified by DNA extraction, PCR, Cloning and sequencing. Results All published studies identified that the 27bp repeat polymorphism in intron 4 was biallelic ( a and b alleles ). Our study found a third allele-allele c, with six 27bp repeats. It was also demonstrated that there was no significant difference in overall allele distribution among the three main ethnic groups and the intron 4 polymorphism didn't show any association with diabetic nephropathy. Conclusion The intron 4 polymorphism was triallelic. We failed to demonstrate any association between this polymorphism with diabetic nephropathy.

Objective To explore the value of manganese-enhanced MRI in locating the rat visual nuclei.Methods The visual nuclei of thirty-six rats were located by 3 different ways including individual MEMRI locating (group A,n= 1 6),anatomical atlas locating (group B,n=1 6)and direct puncture by using the data obtained in MEMRI (group C,n=4).After unilateral intra-vitreal injection of MnCL2 (30 mmol/L×3 μL)in group A,the brain MRI was performed 24 h later.The location coordinate of lateral geniculate nucleus (LGN) and superior colliculus (SC)were recorded individually.The nuclei injections (3% fluorogold solution,1 μL)were performed by using different location coordinate in groups A and B.The rat’s retinas were examined under fluorescence microscope 5 days later,and the results were compared between the two groups.After brain nucleus puncture injection (30 mmol/L MnCL2 solution,0.5 μL),MRI was performed 1 h later in group C.Results The success rate was 93.8% (1 5/1 6)in group A,and 65.5% (10/1 6)in group B.The difference between groups was statistically significant (P<0.05).All the injection locations of C group were agreed with atlas.Conclusion MEMRI in the visual nucleus stereotactic can improve the accuracy of location.

AIM:To investigate the role of B7 homologue 6 (B7-H6) over-expression in natural killer (NK) cell-mediated hepatocyte apoptosis. METHODS:The full-length fragment of B7-H6 gene was amplified by PCR and sub-cloned into linearized eukaryotic expression vector pIRES2-EGFP to construct recombinant B7-H6 over-expression vector pIRES2-EGFP-B7-H6. The recombinant plasmid was identified by double digestion, PCR and sequencing, and was then transfected into L02 cells. The expression of EGFP was observed by fluorescence microscopy and the transfection efficiency was evaluated by flow cytometry. B7-H6 expression was confirmed by qRT-PCR and Western blot. The L02 cells transfect-ed with pIRES2-EGFP-B7-H6 recombinant plasmid were co-cultured with NK-92 cells at different effector/target ratios,and the cytotoxicity of NK-92 cells was evaluated by CCK-8 assay.RESULTS:The strong green fluorescence in the L02 cells was observed under fluorescence microscope 48 h after transfection. The transfection efficiency reached 92.6%. The ex-pression of B7-H6 at mRNA and protein levels was remarkably increased 48 h after transfection. The cytotoxicity of NK-92 cells against L02 cells transfected with pIRES2-EGFP-B7-H6 plasmid was significantly higher than that of the null vector transfection group (P<0.05).CONCLUSION:The recombinant eukaryotic expression vector pIRES2-EGFP-B7-H6 was constructed successfully. The cytotoxic effect of NK-92 cells against L02 cells can be enhanced by transfecting L02 cells with pIRES2-EGFP-B7-H6 plasmid.

The effect of combined low-frequency repetitive transcranial magnetic stimulation (LF rTMS) and virtual reality (VR) training in patients after stroke was assessed. In a double-blind randomized controlled trial, 112 patients with hemiplegia after stroke were randomly divided into two groups: experimental and control. In experimental group, the patients received LF rTMS and VR training treatment, and those in control group received sham rTMS and VR training treatment. Participants in both groups received therapy of 6 days per week for 4 weeks. The primary endpoint including the upper limb motor function test of Fugl-meyer assessment (U-FMA) and wolf motor function test (WMFT), and the secondary endpoint including modified Barthel index (MBI) and 36-item Short Form Health Survey Questionnaire (SF-36) were assessed before and 4 weeks after treatment. Totally, 108 subjects completed the study (55 in experimental group and 53 in control group respectively). After 4-week treatment, the U-FMA scores [mean difference of 13.2, 95% confidence interval (CI) 3.6 to 22.7, P<0.01], WMFT scores (mean difference of 2.9, 95% CI 2.7 to 12.3, P<0.01), and MBI scores (mean difference 16.1, 95% CI 3.8 to 9.4, P<0.05) were significantly increased in the experimental group as compared with the control group. The results suggested the combined use of LF rTMS with VR training could effectively improve the upper limb function, the living activity, and the quality of life in patients with hemiplegia following subacute stroke, which may provide a better rehabilitation treatment for subacute stroke.
Aged ; Arm ; physiopathology ; Double-Blind Method ; Female ; Humans ; Male ; Middle Aged ; Prospective Studies ; Stroke ; physiopathology ; therapy ; Transcranial Magnetic Stimulation ; Virtual Reality Exposure Therapy

OBJECTIVETo establish a full-quantified fingerprint method for analyzing Chinese herbal additives by HPLC.

METHODA HPLC in combination with PDA detector was applied with a phenomenex luna C18 (4.6 mm x 250 mm, 5 microm) column by gradient elution using acetonitrile-0.1% formic acid solution as the mobile phase. The flow rate was 1 mL x min(-1) and the detection wavelength was set at full spectrum scan.

RESULTAccording to the selected chromatographic conditions, the full-quantified fingerprint of the Chinese herbal additive has good precision, reproducibility, stability and recovery.

CONCLUSIONThe HPLC method developed for simultaneous determination of seven compounds is simple and valid. It can be used for quality evaluation and quality control of Chinese herbal additive and its processing products.

Chemistry, Pharmaceutical ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis

AIMTo investigate the effects of Angelica sinensis on the expression of Flt-1, Flk-1 mRNA after the ischemic brain injury in rats.

METHODSWistar rats randomly divided into two groups: group A rats underwent middle cerebral artery occlusion (MCAO) for 2 hours by suture, group B rats underwent MCAO for 2 hours meanwhile received treatment with Angelica sinensis (5g/kg). At 1 st d, 3 rd d and 7 th d after reperfusion, 36 rats( n = 18 in each group) were assessed by neurological scale and brain tissue was taken to assess the lesion ration with 2, 3, 5-triphenyltetrazolium chloride (TTC) staining. The other rats (n = 3 at different time points in each group) were decapitated at 3 h, 6 h, 12 h , 1 st d, 3 rd d, 7 th d after reperfusion. Quantitative reverse transcription and polymerase chain reaction (RT-PCR) technique was used to examine the gene expression of Flt-1, Flk-1.

RESULTSThe neurologic deficit score of rats in group B decreased significantly compared with group A at the same time point (P < 0.05). The infarct volume of group A was significant greater than group B at the same time point after reperfusion (P < 0.01). The results of RT-PCR revealed that the gene expression of Flt-1, Flk-1 in the two groups increased from 3 h after reperfusion and reached its peak at the time of 3 rd d after reperfusion, then declined gradually. The gene expression of Flt-1, Flk-1 in the group B was significantly increased than group A at the same time point (P < 0.01). The gene expression of Flk-1 was positive correlated with Flt-1 in two groups (r = 0.957).

CONCLUSIONThe increasing amount of Flt-1, Flk-1 expression was enhanced by Angelica sinensis following transient interruption of cerebral blood flow in rats.

Angelica sinensis ; Animals ; Brain Ischemia ; metabolism ; Male ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar ; Reperfusion Injury ; metabolism ; Vascular Endothelial Growth Factor Receptor-1 ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; metabolism

Nanostructured materials are gaining new impetus owing to the advancements in material fabrication techniques and their unique properties (their nanosize, high surface area-to-volume ratio, and high porosity). Such nanostructured materials mimic the subtleties of extracellular matrix (ECM) proteins, creating artifi cial microenvironments which resemble the native niches in the body. On the other hand, the isolation of mesenchymal stem cells (MSCs) from various tissue sources has resulted in the interest to study the multiple differentiation lineages for various therapeutic treatments. In this review, our focus is tailored towards the potential of biomimetic nanostructured materials as osteoinductive scaffolds for bone regeneration to differentiate MSCs towards osteoblastic cell types without the presence of soluble factors. In addition to mimicking the nanostructure of native bone, the supplement of collagen and hydroxyapatite which mimic the main components of the ECM also brings signifi cant advantages to these materials.
Biomimetics ; instrumentation ; methods ; Bone Transplantation ; Collagen Type I ; Extracellular Matrix ; Humans ; Mesenchymal Stromal Cells ; Nanostructures ; Osteogenesis ; Tissue Engineering ; instrumentation ; methods

Most important international guidelines recommend the use of CHA 2 DS 2 -VASc and HASBLED scores for stroke and bleeding risk assessments in atrial fibrillation (AF) patients, respectively. The 2020 AF guidelines of European Society of Cardiology have revised the definition of “C: congestive heart failure (HF)” component, and now patients with either HF with reduced ejection fraction or preserved ejection fraction should be assigned 1 point.Hypertrophic cardiomyopathy was also included. Besides, the revised “V: vascular diseases” component included both prior myocardial infarction and “angiographically significant coronary artery disease”. It is important to understand that the stroke and bleeding risks of AF patients were not static and should be re-assessed regularly. A high HAS-BLED score itself should not be the only reason to withhold or discontinue oral anticoagulants, but remind physicians for the corrections of modifiable bleeding risk factors and more regular follow up. In the future, the AF duration and left atrial function may play an important role for personalized evaluation of individual stroke risk while more studies are necessary.