Objective To study the effects of prokaryotic ubiquitin-like protein ( Pup)-proteasome system on the growth of Mycobacterium strains.Methods The genes encoding Pup ( pup gene) and protea-someβsubunit ( prcB gene) were respectively knocked out from Mycobacterium smegmatis ( M.sm) strains by homologous recombination.The growth and viability of the wild-type and mutant strains of M.sm were an-alyzed under normal culture condition and under hypoxia as well as anaerobic conditions.Results The pup and prcB genes were completely and precisely knocked out from M.sm strains and the mutant strains were named △SM-Pup and△SM-prcB, respectively.The△SM-Pup strains grew faster than the wild type ( WT) and△SM-prcB strains.No significantly differences in the growth of M.sm were found between the WT and△SM-prcB strains.Conclusion The Pup-proteasome system was involved in the growth of M.sm, espe-cially the pup gene.There was difference between pup and prcB genes in regulating the growth of M.sm.The functions and influences of Pup-proteasome system still need further investigation.

AIM:To observe and compare clinical effects of coaxial 1. 8mm microincision phacoemulsification and 3. 2mm small incision phacoemulsification.
●METHODS:A total of 117 eyes of 85 patients with age-related cataract in our hospital were divided randomly into two groups:43 patients (59 eyes) in the coaxial 1. 8 mm microincision cataract surgery group ( C - MlCS ) , 42 patients (58 eyes) in the coaxial 3. 2 mm traditional small incision cataract surgery group (C-SlCS). A total of 117 eyes were received phacoemulsification with intraocular lens implantation. Uncorrected visual acuity was recorded preoperatively and postoperatively at 1, 7, 30 and 90d. The effective phacoemulsification time and average ultrasound energy were recorded in surgery. Corneal endothelial cell and corneal topography were recorded preoperatively and postoperatively at 90 d.
●RESULTS:Uncorrected visual acuity ( logMAR) was no overall statistical significance difference between C-MlCS group and C-SlCS group (P>0. 05), but was significant statistical difference in different time-point within both groups(P<0. 05). Uncorrected visual acuity in different time-point had nothing to do with corneal wound size in cataract surgery(P>0. 05). On the 1 day after surgery, uncorrected visual acuity was 0. 16±0. 11 in C-MlCS group and 0. 22±0. 18 in C-SlCS group(P<0. 05). AVE was (7. 00± 2.72)% in C-MlCS group and (6. 16±3. 16)% in C-SlCS group (P>0. 05). EPT was (3. 09±1. 61)s in C-MlCS group and (3. 20±1. 92)s in C-SlCS group (P>0. 05). At 90 d after surgery, corneal endothelial cell loss percentage was (5. 81±2. 28)% in C-MlCS group and (5. 69±2. 38)% in C-SlCS group (P>0.05), SlA was (0.35±0.11) Din C-MlCS group and (0. 61±0. 13) D in C-SlCS group (P<0. 05).
● CONCLUSION: Compared with coaxial 3. 2mm traditional small incision cataract surgery, 1. 8mm coaxial microincision cataract surgery can get earlier visual rehabilitation and significantly reduce SlA. The coaxial 1. 8mm microincision cataract surgery is safe, effective and deserves further clinical applications.

Animals ; Cholestasis ; complications ; Fibrosis ; etiology ; Gallbladder ; pathology ; Humans ; Kidney ; pathology ; Lipid Metabolism ; Liver Cirrhosis ; etiology ; therapy ; Myocardium ; pathology ; Receptors, Cytoplasmic and Nuclear ; physiology

A fast and sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method for the determination of prodrug of paclitaxel (Pro-PTX) and paclitaxel (PTX) in rat plasma was developed. The plasma samples were subjected to protein precipitation with acetonitrile (0.1% formic acid), and then separated by LC with an Ultimate AQ-C18 column (50 mm × 3.0 mm, 3 μm) and acetonitrile-1 mmol·L^-1 ammonium formate (containing 0.1% formic acid) as the mobile phase. Multiple reaction monitoring (MRM) scanning mode was used to detect the ion responses m/z 983.4→415.2 (Pro-PTX), m/z 854.4→286.1 (PTX) and m/z 808.3→527.2 (Docetaxel, internal standard) by using a triple quadrupole tandem mass spectrometer with electrospray ionization source and positive ion mode. The method validation results show that the linear ranges of Pro-PTX and PTX in plasma were 2.00-500 ng·mL^-1 (r > 0.99) and 4.00-1 000 ng·mL^-1 (r > 0.99), the lower limit of quantification was 2.00 ng·mL^-1 and 4.00 ng·mL^-1, respectively; the quality control samples with low, medium and high concentrations of Pro-PTX and PTX were within the batch, the relative standard deviation (RSD) between batches were all less than 9%; the relative deviation (RE) was within the range of ± 9%; In the stability test, both Pro-PTX and PTX in plasma were stable under various storage conditions. The method was sensitive, specific, and reproducible, and was suitable for the pharmacokinetic study of Pro-PTX in rats. Animal experiments were approved by the Ethics Committee of Laboratory Animal Management and Animal Welfare, Institute of Materia Medica, Chinese Academy of Medical Sciences (No.: 00003552).

Aim To investigate the effects of dioscin ( Dio) on rat myocardial contractility. Methods Left ventricular contractile function was measured using the Langendorff non-recirculating mode of isolated rat heart perfusion. Effects of low, middle and high concentra-tion of Dio were investigated by measuring left ventricu-lar systolic pressure ( LVSP ) and left ventricular end diastolic pressure ( LVEDP) . Also, peak rates of rise/fall of left ventricular pressure ( ± dp/dtmax ) of isolated rat heart were calculated. Effects of Dio on intracellu-lar free calcium concentration in rat H9 c2 cells were measured by using the confocal microscopy. Mitochon-drial membrane potential was detected with multifunc-tional microplate reader. Results With 0. 1, 1 μmol · L-1 Dio, LVSP were significantly enhanced from (11. 55 ± 0. 52), (10. 53 ± 0. 28) kPa to (13. 08 ± 0. 72), (12. 53 ±0. 64) kPa(P<0. 01); +dp/dtmax were dramatically increased from ( 0. 38 ± 0. 10 ) , (0. 40 ± 0. 07) kPa·ms-1 to (0. 42 ± 0. 11), (0. 43 ± 0. 02) kPa·ms-1(P<0. 05). With the 10μmol· L-1 Dio, LVSP and + dp/dtmax were both decreased from (12. 13 ± 0. 33) kPa and (0. 42 ± 0. 04) kPa· ms-1 to ( 9. 46 ± 0. 77 ) kPa and ( 0. 24 ± 0. 04 ) kPa ·ms-1 (P <0. 01). With 0. 1, 1, 10 μmol·L-1 Dio, the relative fluorescence intensity of intracellular free calcium concentrations was increased significantly from (16. 62 ± 0. 89) to (21. 48 ± 0. 80), (25. 68 ± 0. 69) and (19. 84 ± 0. 66)(P <0. 01)respectively. 0. 1, 1μmol·L-1 Dio showed no significant effects on the mitochondrial membrane potential of rat H9 c2 cells, while with effects of 10 μmol·L-1 Dio, the ra-tio of JC-1 monomer and J-aggregates was changed from (1. 14 ± 0. 03) to (1. 35 ± 0. 06)(P<0. 01), indica-ting a decrease in the mitochondrial membrane poten-tial. Conclusion Low and middle concentrations of Dio show a positive inotropic effect on isolated rat heart, as the LVSP and + dp/dtmax are enhanced, which may concern with the increase of the intracellu-lar concentration of Ca2+. It will not cause the calcium overload while the intracellular concentration of Ca2+ is increased by low and middle concentration of Dio in the myocytes except high concentration of Dio.

Objective To understand the radiation protection effect of pre-irradiation administrations of nilestriol on the mice with bone marrow type of acute radiation syndrome after irradiation with 60Co γ-rays,along with its mechanisms for improvement of hematopoiesis.Methods The nilestriol administration protocols were prepared by analysis of peripheral blood cell counts and survival rate experiment on mice.The mechanisms by which the pre-irradiation twice administrations improved the post-irradiation recovery of bone marrow hematopoiesis were studied by the analysis of the surface marker of bone marrow hematopoietic stem/progenitor cells of mice and by the inspection of hematopoietic progenitor cell colony and by using histopathological assessment of bone marrow.Results Pre-irradiation administration of nilestriol at two-or three-day intervals had been shown to increase survival rates up to 100％ in mice exposed to 9.0 Gy γ-rays,which was superior to a single administration (20％,x2 =21.66,21.66,P ＜0.05).The pre-irradiation administration both at one-day or two-day intervals were capable of improving the recovery of peripheral blood counts,including white blood cell (WBC),red blood cell (RBC),and platelet in mice exposed to 6.5 Gy (F =21.33,100.9,49.34,19.19,P ＜ 0.05),showing the better effects than a single administration (F =17.11,63.38,21.89,14.37,P ＜ 0.05).The two-day-interval administration of nilestriol could significantly increase the numbers of bone marrow hematopoietic stem/progenitor cell counts (t =8.58,2.80,P ＜ 0.05) in mice on day 10 after 6.5 Gy irradiation.This also could be capable to significantly improve colony formation,with there being statistical difference compared with single administration(t =4.29,6.34,P ＜ 0.05).Also the administration at two-day-interval were also usefull in reconstruction of hematopoietic cell hyperplasia of bone marrow of irradiated mice.Conclusions As compared with conventional single admination,the pre-irradiation multiple administrations of nilestriol showed significantly improved radiation protection effects.Considering a nuclear medical emergency rescue,it is recommended to follow the pre-irradiation administration of nilestriol at two-day interval,which could obtain the best protection effects at minimum administration frequency.

The T-cell immunoglobulin and immunoreceptor tyrosine-based inhibitory motif domain (TIGIT) is an inhibitory receptor mainly expressed on active T-cells, or natural killer cells (NK cells) that activate negative stimulus signals in immune cells by combining with multiple ligands on the surface of target cells including tumor cells and infected cells. TIGIT plays an important regulatory role in the immune pathogenesis of tumors, viral infections and various autoimmune diseases by inhibiting the over activation of cells and the over secretion of proinflammatory cytokines. Recent researches show that TIGIT is highly expressed in T cells and NK cells of cancer patients, and is related to disease progression and poor clinical prognosis. Researchers try to enhance the activity of T cells or NK cells by blocking the binding of TIGIT and its ligand for therapeutic intervention. At present, there have been many reports about the use of anti-TIGIT monoclonal antibody treatment in different mouse tumor models leading to tumor regression, TIGIT has received extensive attention in cancer immunotherapy as a promising target for next generation cancer immunotherapy. Several clinical trials are currently evaluating the efficacy of anti-TIGIT monoclonal antibodies (mAbs) in patients with several cancers. The most advanced candidate, tiragolumab, has exhibited remarkable efficacy in programmed cell death ligand 1 (PD-L1)-positive non-small cell lung carcinoma (NSCLC) patients in phase Ⅱ clinical trials, in combination with PD-L1 blockade. However, the specific mechanism of TIGIT blockade remains to be fully elucidated.

Objective To explore the ameliorative effects of standardized tertiary rehabilitation(STR)on the quality of life(QOL)of stroke patients with hemiplegia.Methods Eighty-two stroke patients were randomly divided into two groups:a rehabilitation group(R group)and a control group.All patients were given routine inter- nal medicine treatment,but STR was also administered to the R group.The QOL of all of the patients was assessed with the brief scale of quality of life(QOL-BREF)at the beginning,and at the end of the Ist month,3rd month and 6th month after stroke.Results There were two deaths in the control group,but no deaths in the R group.At ever- y stage,the R group returned better scores for physiological health,psychological state,social and environment rela- tionships,the subjective QOL and health items,and in comprehensive self-evaluation.QOL scores,except for social relationships,were consistently,significantly higher in the R group.The R group's QOL scores improved obviously in the first 3 months after stroke,and then more slowly in the next 3 months.Conclusions STR markedly improved the QOL of stroke patients.Their QOL scores improved obviously soon after stroke,but slowly later on.

Objective To observe the effects of standardized three stage rehabilitation treatment on the neu- rological deficit scores (NDS) and ADL performance of ischemic stroke patients.Methods A total of 164 ischemic stroke patients were recruited and randomly divided into a rehabilitation group and a control group.The neurological function and ADL performance of the patients were assessed by using NDS and Modified Barthcl Index (MBI) at the admission,at the end of 1st,3rd and 6th months post stroke.Results No significant differences were found be- tween the rehabilitative and the control groups with regard to NDS and MBI at admission.The NDS demonstrated a decreasing tendency,while the MBI score an increasing tendency in both groups.In the control group,significant difference of NDS was found between admission and the end of 1st month as well as between the end of the 1 st and the 3rd months.In rehabilitation group,significant difference was revealed between all the time points with regard to NDS and MBI scores.At the end of the 1st,3rd and 6th months,the MBI scores of the rehabilitation group were signifi- cantly higher than those of the control group,indicating that the ADL performance of those treated with standardized three-stage rehabilitation protocol was improved quicker than those without the protocol.Conclusion Standardized three-stage rehabilitation treatment could improve the neurological function and ADL performance of the ischemic stroke patients.

Objective To examine the reliability,validity and sensitivity of FCA for stroke patients. Methods A total of 82 patients with stroke were divided into a treatment group and a control group randomly.The patients in the control group were given routine internal medicine treatment,while those in the treatment group were given standardized rehabilitation therapy in addition to the routine internal medicine treatment.All patients were as- sessed with FCA and Modified Barthel Index(MBI),at admission and the end of the 1st month,3rd and 6th month,respectively after the onset of stroke.The internal consistency,split-half reliability,content validity,crite- ria validity and sensitivity of FCA were tested.Repeat measure ANOVA and correlation analysis were used in the statistical analysis.Results There existed a good internal consistency(Cronbach?=0.901～0.973)as well as a good split-half reliability(R=0.949～0.981)of FCA.There was a medium to strong correlation among the 18 items scores of the FCA and FCA scores(P