Objective:To explore the effect of NOD-like receptor thermal protein 3 ( NLRP3) knockout in γ-aminobutyric acid (GABA)-ergic neurons in the hippocampal CA1 area on improving cognitive dysfunction in mice after traumatic brain injury (TBI). Methods:Forty-eight healthy male NLRP3 flox/flox mice weighing 25-28 g were randomly divided into 4 groups ( n=12): sham-operated+control virus group (SV group), sham-operated+ NLRP3 specific knockout group (SG group), TBI+control virus group (TV group), TBI+ NLRP3 specific knockout group (TG group). TBI in the TV and TG groups was established by free-fall method, while surgical procedures such as scalp incision and cranial window opening without impact were given to the SV and SG groups. Adenovirus was injected into the hippocampal CA1 area of SG and TG groups 21 d before TBI to induce NLRP3 specific knockout in GABA-ergic neurons in the hippocampal CA1 area; empty virus was injected into the CA1 area of SV and TV groups. Cognitive function was evaluated using novel object recognition test 30 and 31 d after TBI, and learning and memory functions were assessed using Morris water maze test 32-36 d after TBI. Field potentials in the hippocampal CA1 area were recorded during novel object recognition 31 d after TBI. After behavioral tests, these mice were sacrificed. Immunofluorescent staining was used to detect the fluorescent intensity of microtubule-associated protein2 (MAP2), glutamic acid decarboxylase 67 (GAD67), and postsynaptic density protein 95 (PSD95) in the hippocampal CA1 area, as well as percentage of pyroptosis-associated inflammatory factor interleukin-18 (IL-18)/GAD67 double-positive neurons in total GAD67 positive neurons. Results:Compared with the SV and SG groups, the TV and TG groups had decreased novel object recognition index, decreased number of platform crossings during the experimental period, increased escape latency on day 3 and day 4 of the training period in Morris water maze test, decreased θ and γ oscillation power in the hippocampal CA1 area during novel object recognition, decreased fluorescent intensity of MAP2, GAD67, and PSD95 in the hippocampal CA1 area, increased percentage of IL-18/GAD67 double-positive neurons, with significant differences ( P<0.05). Compared with the TV group, the TG group had increased novel object recognition index, increased number of platform crossings in Morris water maze test, decreased escape latency during the training period, increased θ and γ oscillation power in the hippocampal CA1 area during novel object recognition, increased fluorescence intensity of MAP2, GAD67, and PSD95 in the hippocampal CA1 area, decreased percentage of IL-18/GAD67 double-positive neurons, with significant differences ( P<0.05). Conclusion:Specific inhibition of NLRP3 expression in GABA-ergic neurons in the hippocampal CA1 area can improve cognitive dysfunction in mice after TBI, whose mechanism may be related to inhibited GABA-ergic neuronal pyroptosis in the hippocampal CA1 area.

Objective To investigate the clinical value of Mycobacterium tuberculosis/nontuberculous mycobacteria(MTB/NTM)nucleic acid detection in cerebrospinal fluid for the diagnosis of tuberculous meningitis(TBM).Methods The clinical data of 120 pa-tients with TBM were collected.Patients were divided into two groups using a random number table method:TBM A group(n=67,cere-brospinal fluid routine,biochemical,Roche culture,tuberculous smear,MTB/NTM nucleic acid detection)and TBM B group(n=53,cerebrospinal fluid routine,biochemical,Roche culture,tuberculous smear,GeneXpert MTB/RIF detection),and another 18 cases of cerebrospinal fluid from patients with other central nervous system infections were collected as control group(C group,cerebrospinal fluid routine,biochemical,Roche culture,tuberculous smear,MTB/NTM nucleic acid,GeneXpert MTB/RIF detection.Results The posi-tive rates of Roche culture,tuberculosis smear and MTB/NTM nucleic acid detection of cerebrospinal fluid in TBM A group were 13.43％(9/67),10.45％(7/67)and 46.27％(31/67),respectively.The positive rates of Roche culture,tuberculosis smear and GeneXpert MTB/RIF detection of cerebrospinal fluid in TBM B group were 15.09％(8/53),11.32％(6/53)and 47.17％(25/53),respective-ly.Among the cerebrospinal fluid GeneXpert MTB/RIF positive cases,lifampin resistance gene was positive in 2 patients.In the C group,patients had negative result in Roche culture,tuberculous smear,MTB/NTM nucleic acid detection and GeneXpert MTB/RIF detection.Conclusion MTB/NTM nucleic acid detection and GeneXpert MTB/RIF detection are useful in diagnosing TBM at early stage.MTB/NTM nucleic acid detection more economical than GeneXpert MTB/RIF detection,and can identify non-tuberculous mycobacteria(NTM),and timely reduce the misdiagnosis of NTM disease,which is worthy of clinical promotion.

Aim To investigate the relationship between triglyceride-glucose(TyG)index and the risk of hy-pertension in middle-aged and older adults in China,and to provide scientific basis for the prevention and treatment of hypertension.Methods Data were obtained from the China health and retirement longitudinal study(CHARLS)in 2011.A multi-stage stratified sampling method was used to select participants.Restricted cubic spline regression model was used to analyze the dose-response relationship between TyG index and the risk of hypertension.Multivariate unconditional Logistic regression model was used to evaluate the association between TyG index and the risk of hyperten-sion.Results A total of 9 987 subjects were included in the analysis,with an average age of(59.16±9.43)years,including 4 707 males(47.13％).The restricted cubic spline regression model showed that the risk of hypertension in-creased with the elevation of TyG index,and there was a linear association(overall association test P＜0.000 1,non-linear association test P=0.201 9).The results of multivariate Logistic regression model showed that compared with Q1(TyG index＜8.23),the OR(95％CI)of hypertension with Q2(8.23≤ TyG index＜8.59),Q3(8.59 ≤ TyG index＜9.04)and Q4(TyG index≥9.04),were 1.09(0.95～1.26),1.53(1.33～1.76)and 1.77(1.52～2.06),respectively.Conclusions With the increase of TyG index,the risk of hypertension gradually increased.TyG index may be an inde-pendent risk factor of hypertension.

Objective:To explore the clinical features, etiologies and outcomes of unknown origin fever after simultaneous pancreas-kidney transplantation(SPK).Methods:From March 2015 to January 2020, clinical data were retrospectively reviewed for 120 SPK recipients.According to the definite evidence of fever, such as microbial culture, imaging findings or rejection, they were divided into three groups of free-fever(FF, n=41)and defined-fever(DF, n=47)and fever of unknown origin(FUO, n=32). The differences in general clinical features, surgical complications, laboratory tests and prognoses were compared.Logistic regression was employed for analyzing the risk factors of FUO and Kapla-Meier for survival analysis.And P<0.05 was deemed as statistically significant. Results:Multivariate analysis revealed that preoperative diabetic gastroenteropathy was an independent risk factor for unexplained fever.Significant differences existed between FUO and DF groups in leucocyte count[6.50(5.13, 7.36)vs.10.36(6.11, 12.97)×10 9/L], C-reactive protein(CRP)[11.75(6.25, 16.85)vs.35.00(16.30, 75.00)μg/ml], procalcitonin[0.13(0.06, 0.18)vs.0.19(0.11, 1.05)ng/ml]( P<0.001, P<0.001, P=0.025). As compared with DF group, 19 recipients in FUO group only received 1-2 antibiotics and there was a shorter course of treatment[13(40.6%)vs.32(68.1%), P=0.016]. For 6(18.7%)recipients after a diagnosis of FUO, clinical outcome was achieved with only NSAIDs.Length of stay was(48.72±19.51)days in FUO group versus(57.36±27.46)days in DF group and the difference was statistically significant( P<0.001). Hospitalization expenses of two groups were 253 463.25 and 334 605.96 yuan respectively and the difference was also statistically significant( P=0.002). Conclusions:Diabetic gastroenteropathy is an independent risk factor for early FUO after SPK transplantation.Inflammatory markers of leukocytes, CRP and procalcitonin in FUO patients are significantly lower than DF group.And these clinical features can help diagnose FUO in an early stage.

Bipolar Disorder ; blood ; immunology ; metabolism ; Depressive Disorder ; blood ; immunology ; metabolism ; Humans ; Proteomics

AIM:To explore the effects of confluent endothelial progenitor cells (EPCs) derived from young and aged rats on the phenotype conversion, proliferation and migration of vascular smooth muscle cells ( SMCs) .METH-ODS:Mononuclear cells were obtained from the bone marrow of young (1~2 month old) and aged (19 to 26 month old) Sprague-Dawley rats and cultured with medium DMEM/F12 ( containing 15% fetal bovine serum, endothelial cell growth supplements (ECGs) 100 g/L, 1 ×105 units/L of penicillin and streptomycin, respectively).EPCs were characterized as double positive for DiI-Ac-LDL uptake and lectin binding.Abdominal aorta was obtained from 1 to 2 month old Sprague-Dawley rats.Vascular SMCs were cultured by tissue explant method and identified byα-SM-actin immunofluorescence.In transwell co-culture system, the confluent EPCs located in the upper chamber and SMCs were seeded on the lower cham-ber.The experiments were divided into passage 3 SMCs group (P3), passage 4 SMCs group (P4), passage 4 SMCs co-culture with EPCs derived from young rats group (P4YE) and passage 4 SMCs co-culture with EPCs derived from aged rats group (P4AE).The protein expression ofα-SM-actin and osteopontin was detected by Western blotting.[3H]-TdR incor-poration assay was used to determine the proliferation.SMC migration was analyzed by scratch wound healing assay.RE-SULTS:Compared with P3 group,α-SM-actin expression in P4 group significantly decreased and osteopontin protein ex-pression obviously increased, whereas no significant change was found in P4YE group.Compared with P4 group, confluent EPCs derived from young and aged rats both markedly increased α-SM-actin and decreased osteopontin expression in P4 SMCs.Compared with aged rat-derived EPCs, young rat-derived EPCs were more effectively to induce a delayed SMC phe-notype transition (from contractile phenotype to a synthetic phenotype), and to inhibit SMC proliferation and migration. CONCLUSION:Co-culture of confluent EPC induces a delayed vascular SMC phenotype transition and inhibits SMCs pro-liferation and migration.Young rat derived EPCs are more effective to induce a delayed vascular SMC phenotype transition and has stronger inhibitory effects on SMCs proliferation and migration compared with that derived from aged rats.

Objective To investigate early renal damage of chronic hepatitis B (CHB) patients and the risk factors related to their renal function. Methods CHB patients who visited the second people’s hospital but did not receive systemic treatment were enrolled in our study. Those who visited for general check-up with no hepatic findings during the same period were selected as control group. Glomerular filtration rate (GFR) of all the participants were estimated by simplified MDRD equation and CKD-EPI equation (designated as M-eGFR and C-eGFR respectively). Influence factors of eGFR were statistically analyzed. Results In the total 528 cases in CHB group, 88 (16.67%) and 62 (11.74%) suffered declined M-eGFR and C-eGFR respectively. By contrast, 10 (8.77%) and 6 (5.26%) cases in the total 114 cases in control group present declined M-eGFR and C-eGFR ac?cordingly. Percentages of renal function impairment, estimated by both M-eGFR and C-eGFR, were higher in the CHB group than those in control group. The difference was statistically significant (χ2=4.518, P<0.05;χ2=4.156, P<0.05). Multiple linear regression analysis indicated that age, HBsAg and body mass index (BMI) were risk factors of M-eGFR while age, HBsAg, gender and serum albumin were risk factors of C-eGFR. On the other hand, HBV-DNA and HBeAg were not risk factors for M-eGFR or C-eGFR. Conclusion HBV infection can lead to early renal damage. Age and HBsAg are main risk factors of renal function impairment. Therefore, renal function should be scrutinized in CHB patients.

OBJECTIVETo investigate the relationship of the mutational status of the ND4 gene and the clinical features of acute myelogenous leukemia (AML) patients with ND4 mutations.

METHODSUsing PCR combined with directly sequencing, we identified somatic mutations of ND4 in 121 primary AML patients to couple with their clinical features.

RESULTSThere were 58 male patients and 63 female patients (median age 49 years, 10-86 years). Eight of 121 patients (6.6%) with de novo AML were found harboring missense mutation of ND4 gene, including 3 patients with A131V (3/8, 37.5%), 2 patients with A404T (2/8, 25%), 1 patient with F149L (1/8, 12.5%), 1 patient with G242D (1/8, 12.5%) and 1 patient with Y409H (1/8, 12.5%), respectively. Patients with ND4 mutations were associated with good karyotype (P=0.049), regardless of gender, age, white blood cell, hemoglobin, platelet, blast cells of bone marrow or immunophenotype (P>0.05). There were no statistical significance in mutations of FLT3-ITD, NPM1, CEBPA, c-KIT and DNMT3A between patients with ND4 mutation and wild-type (wt) ND4 (P>0.05). The median overall survival of patients with ND4 mutations and wt ND4 were all not reached. The median relapse-free survival were not reached and 29(2-53) months, respectively (P>0.05). There was no significance in the ratio of CR and RR patients between wt ND4 and ND4 mutated groups (P>0.05).

CONCLUSIONIt was concluded that novel ND4 mutations could be found in de novo AML patients, especially in patients with good karyotype. Thus, ND4 mutations might play an important role in AML prognosis. However, whether the mitochondria dysfunction contribute to leukemogenesis needs to be further investigated.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Female ; Humans ; Leukemia, Myeloid, Acute ; drug therapy ; genetics ; Male ; Middle Aged ; Mutation ; NADH Dehydrogenase ; genetics ; Prognosis ; Young Adult

Research on novel pullulanase has major significance on the domestic industrialization of pullulanase and the breakdown of foreign monopoly. A thermophilic bacteria LM 18-11 producing thermostable pullulanase was isolated from Lunma hot springs of Yunnan province. It was identified as Anoxybacillus sp. by 16S rDNA phylogenetic analysis. Full-length pullulanase gene was cloned from Anoxybacillus sp. LM18-11. The optimum temperature of the pullulanase was between 55 and 60 degrees C with a half-life as long as 48 h at 60 degrees C; and its optimum pH was between 5.6 and 6.4. V(max) and K(m) of the pullulanase was measured as 750 U/mg and 1.47 mg/mL, which is the highest specific activity reported so far. The pullulanase crystals structure showed a typical alpha-amylase family structure. The N-terminal has a special substrate binding domain. Activity and substrate binding were decreased when the domain was deleted, the V(max) and K(m) were 324 U/mg and 1.95 mg/mL, respectively. The pullulanase was highly heterologous expressed in Bacillus subtilis by P43 promoter. The extracellular enzyme activity was 42 U/mL, which increased more than 40 times compared to the initial strain. This pullulanase has good application prospects.
Anoxybacillus ; classification ; enzymology ; China ; Glycoside Hydrolases ; metabolism ; Hydrogen-Ion Concentration ; Phylogeny ; RNA, Ribosomal, 16S ; genetics ; Temperature

Objective To study the potential role of PKC inhibitor Rottlerin in the treatment of asthma. Methods Balb/c mice were sensitized and challenged with ovalbumin(OVA) protein to construct murine experimental model of asthma. Then the mice were treated by means of injection of Rottlerin or phosphate buffered saline (PBS) into the abdominal cavity. Changes of the total serum IgE, pulmonary eosinophils, cytokines and pulmonary inflammation were investigated. Results Injection of Rottlerin at the dose 0.3 mg/kg into the murine abdominal cavity could inhibit the infiltration of pulmonary eosinophils and pulmonary inflammation and significantly decrease the production of total serum IgE and Th 2 cytokines as well. Conclusion Rottlerin can inhibit murine experimental asthma.