Objective To investigate the clinical efficacy of acupuncture at gastrointestinal Back-Shu and Front-Mu points for post-stroke functional constipation. Method Sixty-six patients with post-stroke functional constipation were randomized to treatment and control groups, 33 cases each. The treatment group received acupuncture at gastrointestinal Back-Shu and Front-Mu points: Weishu (BL21), Dachangshu (BL25), Xiaochangshu (BL27), Zhongwan (CV12), Tianshu (ST25) and Guanyuan (CV4). The control group took Chinese herbal medicine Liumo Decoction prescribed originally. The clinical therapeutic effects were compared between the two groups. The CCS score was recorded and fecal characteristics were scored in the patients at baseline, during treatment and at follow-up. Result The total efficacy rate was 90.9％ in the treatment group and 42.4％ in the control group. The CCS score and the fecal characteristic score were lower in the two groups after treatment compared with before (P<0.05) and the therapeutic effect was more marked in the treatment group (P<0.05). The CCS score and the fecal characteristic score increased in the two groups at follow-up, but the increments were significantly smaller in the treatment group than in the control group (P<0.05). Conclusion Shu-Mu point combination acupuncture can significantly relieve the symptoms of post-stroke functional constipation and change fecal characteristics. It is superior to Chinese herbal treatment alone.

Objective To analyze the risk factors of patients with trauma in intensive care unit (ICU), a new warning scoring system is established for predicting the incidence of sepsis in traumatic patients; and to provide a new simple method of clinical score, which could provide a reference for clinical prevention and treatment of sepsis. Methods The clinical data of 591 patients with trauma in the ICU of the Army Specialized Medical Center of Army Medical University and Affiliated Hospital of Zunyi Medical University from January 2012 to December 2017 were retrospectively analyzed. The patients were divided into sepsis group (n = 382) and non-sepsis group (n = 209) according to their clinical outcome. The basic clinical data of all ICU trauma patients were collected, and the differences in gender, age, underlying diseases, and vital signs, critical illness scores, blood culture results and laboratory biochemical examinations within 24 hours of ICU admission between the two groups were analyzed. Univariate Logistic regression analysis was used to screen the related factors leading to sepsis. The indexes with P < 0.12 analyzed by univariate Logistic regression analysis were included in multivariate Logistic regression analysis. The risk factors of sepsis in traumatic patients were screened and assigned, and the total score was sepsis early warning score. The receiver operating characteristic (ROC) curve was plotted to evaluate the predictive value of the warning score of sepsis in patients with trauma. Results The incidence of sepsis in ICU trauma patients was 64.6% (382/591), and the ICU mortality was 10.5% (40/382). The traffic accident was a common cause of ICU trauma patients. Compared with non-sepsis patients, Glasgow coma score (GCS), proportion of past history, red blood cell (RBC), platelet (PLT), albumin (Alb) were lower in patients with sepsis, and body temperature, pulse, acute physiology and chronic health evaluation Ⅱ (APACHEⅡ), sequential organ failure assessment (SOFA), injury severity score (ISS), new injury severity score (NISS), fraction of inspired oxygen (FiO2), blood sodium, activated partial thromboplastin time (APTT), prothrombin time (PT), procalcitonin (PCT), C-reactive protein (CRP) levels were higher, blood transfusion, central venous catheterization, mechanical ventilation, shock, multiple organ dysfunction syndrome (MODS), open injury and multiple injuries were more common, the duration of mechanical ventilation, ICU days and total hospital days were longer, and all the differences were statistically significant. Most of the traumatic patients with sepsis were undergone with multiple trauma. Compared with non-sepsis patients, the proportion of multiple position trauma was significantly higher than patients without sepsis. And most traumatic patients were insulted in head, face and neck. The risk factors were screened by univariate and multivariate Logistic stepwise regression analysis, the indexes into the regression model were pulse > 100 bpm [odds ratio (OR) = 1.617, 95% confidence interval (95%CI) = 0.992-2.635, P = 0.044], APTT > 36 s (OR = 2.164, 95%CI =1.056-4.435, P = 0.035), shock (OR = 1.798, 95%CI = 1.056-3.059, P = 0.031), mechanical ventilation (OR = 5.144, 95%CI = 2.302-11.498, P < 0.001), APACHEⅡ > 21 (OR = 3.348, 95%CI = 1.724-6.502, P < 0.001), NISS > 25 (OR = 3.332, 95%CI = 1.154-9.624, P = 0.026), assigning scores were 0.5, 1.0, 0.5, 1.5, 1.5, 1.5, respectively, which were included in the new warning score of sepsis. ROC curve analysis showed that the area under ROC curve (AUC) of warning score for predicting sepsis in patients with trauma was 0.782, which was significantly higher than the APACHEⅡ(AUC = 0.672), APTT (AUC = 0.574) and NISS (AUC = 0.515) with significant difference (all P < 0.01). When the cut-off value of sepsis warning score was 4.0, the sensitivity and specificity were 71.7% and 61.9%, respectively. Conclusions Close monitoring and stabilization of vital signs of traumatic patients within 24 hours of ICU admission and reduction of unreasonable invasive mechanical ventilation time are expected to reduce the incidence of sepsis in traumatic patients. New warning score of sepsis consisted of six factors: pulse, APTT, shock, mechanical ventilation, APACHEⅡ and NISS. Rational use of warning score of sepsis would help us to assess the prognosis of traumatic patients more easily and effectively, and the predicted effect is much better than APACHEⅡ, APTT and NISS.

OBJECTIVE: To investigate the clinical features, accompanying gene mutation characteristics and prognostic factors of adult patients with acute myeloid leukemia with mutated NPM1 (NPM1AML). METHODS: Seventy-three patients with newly diagnosed adult NPM1AML were selected. The mutations of 22 genes were detected by second generation sequencing and 43 fusion genes of AML were detected by real-time fluorescent quantitative PCR. The Kaplan-Meier survival curve and Cox multivariate regression analysis were used to study the prognostic factors. RESULTS: A total of 74 NPM1 site mutations were detected in 73 patients with NPM1AML. The incidence rates were 92.0% L287fs, 2.7% Q289fs and W288fs, 1.4% L258fs and Q289H, among which 1 patient had 2 NPM1 mutations; the different mutation sites had no effect on the prognosis of NPM1AML. The median value of NPM1 variant allele frequency (VAF) was 35.4% (1.8%-56.6%). Based on the uppermost quartile of 38.4%, the patients were classified as NPM1 VAF>38.4% (NPM1AML) and NPM1 VAF≤38.4% (NPM1AML). Compared with NPM1AML, the early mortality rate was statistically significantly higher (33.3% vs 7.3%, P<0.05), and median EFS (148 d，95%CI 58-238 d vs 372 d，95%CI 264-480 d) (P<0.01) and median OS (179 d 95%CI 6-352 d vs 444 d) (P<0.01) were significantly shorter in NPM1 AML. A total of 126 accompanying gene mutation sites were detected in 87.7% of patients with NPM1AML. The patients with NRAS gene mutation displayed a higher rate of complete remission (100% vs 58%) (P<0.05) and longer median OS (not reached to 320 d, 95%CI 150-490 d) (P<0.05). The 43 fusion genes were examined in 65 out of 73 cases of NPM1AML, and in all the patients the fusion gene test was negative. Multivariate analysis showed that NPM1 VAF>38.4% was an independent prognostic factor for EFS (HR=3.1, 95% CI 1.6-6.4, P<0.01) and OS (HR=3.0, 95% CI 1.4-6.2, P<0.01). CONCLUSION The NPM1 gene mutation in AML patients often is accompanied by other gene mutations, while the coexistence of fusion genes is rare; high NPM1 mutant allele burden is an independent prognostic factor for adult AML patients with mutated NPM1.
Alleles ; Humans ; Leukemia, Myeloid, Acute ; genetics ; Mutation ; Nuclear Proteins ; genetics ; Prognosis ; fms-Like Tyrosine Kinase 3

In this study, the synthetic pathway of β-amyrin was constructed in the pre-constructed Saccharomyces cerevisiae chassis strain Y0 by introducing β-amyrin synthase from Glycyrrhiza uralensis, resulting strain Y1-C20-6, which successfully produced β-amyrin up to 5.97 mg·L~(-1). Then, the mevalonate pyrophosphate decarboxylase gene(ERG19), mevalonate kinase gene(ERG12), 3-hydroxy-3-methylglutaryl-CoA synthase gene(ERG13), phosphomevalonate kinase gene(ERG8) and IPP isomerase gene(IDI1)were overexpressed to promoted the metabolic fluxto the direction of β-amyrin synthesis for further improving β-amyrin production, resulting the strain Y2-C2-4 which produced β-amyrin of 10.3 mg·L~(-1)under the shake flask fermentation condition. This is 100% higher than that of strain Y1-C20-6, illustrating the positive effect of the metabolic engineering strategy applied in this study. The titer of β-amyrin was further improved up to 157.4 mg·L~(-1) in the fed-batch fermentation, which was almost 26 fold of that produced by strain Y1-C20-6. This study not only laid the foundation for the biosynthesis of β-amyrin but also provided a favorable chassis strain for elucidation of cytochrome oxidases and glycosyltransferases of β-amyrin-based triterpenoids.
Fermentation ; Glycyrrhiza uralensis ; enzymology ; genetics ; Industrial Microbiology ; Intramolecular Transferases ; genetics ; Metabolic Engineering ; Oleanolic Acid ; analogs & derivatives ; biosynthesis ; Saccharomyces cerevisiae ; metabolism

BACKGROUND: Pulmonary vein (PV) occlusion generally depends on repetitive contrast agent injection when cryoballoon ablation for atrial fibrillation (AF). The present study was to compare the effect of cryoballoon ablation for AF guided by transesophageal echocardiography (TEE) vs. contrast agent injection. METHODS: Eighty patients with paroxysmal AF (PAF) were enrolled in the study. About 40 patients underwent cryoballoon ablation without TEE (non-TEE group) and the other 40 underwent cryoballoon ablation with TEE for PV occlusion (TEE group). In the TEE group during the procedure, PVs were displayed in 3-dimensional images to guide the balloon to achieve PV occlusion. The patients were followed up at regularly scheduled visits every 2 months. RESULTS: No differences were identified between the groups in regard to the procedure time and cryoablation time for each PV. The fluoroscopy time (6.7 ± 4.2 min vs. 17.9 ± 5.9 min, P < 0.05) and the amount of contrast agent (3.0 ± 5.1 mL vs.18.1 ± 3.4 mL, P < 0.05) in the TEE group were both less than the non-TEE group. At a mean of 13.0 ± 3.3 mon follow-up, success rates were similar between the TEE group and non-TEE group (77.5% vs. 80.0%, P = 0.88). CONCLUSIONS Cryoballoon ablation with TEE for occlusion of the PV is both safe and effective. Less fluoroscopy time and a lower contrast agent load can be achieved with the help of TEE for PV occlusion during procedure.
Aged ; Atrial Fibrillation ; diagnostic imaging ; surgery ; Contrast Media ; Cryosurgery ; methods ; Echocardiography, Three-Dimensional ; methods ; Echocardiography, Transesophageal ; methods ; Female ; Humans ; Male ; Middle Aged ; Pulmonary Veins ; diagnostic imaging ; surgery ; Treatment Outcome

OBJECTIVE: To investigate the effects of optical genetic techniques on new neurons through the Wnt/β-Catenin pathway. METHODS: Neural stem cells (ESCs)were extracted from the cerebral cortex of fetal rat and transfected by lentivirus carrying DCX-ChR2-EGFP gene and the expression of DCX of newborn neurons differentiated from neural stem cells were observed. All cells were divided into 3 groups(n=9): control group, NSCs+EGFP and NSCs+ChR2 groups. The control group was normal cultured NSCs (NSCs group); the neural stem cells in NSCs+EGFP group were transfected with lentivirus carrying EGFP gene. The neural stem cells in NSCs+ChR2 group were infected with lentivirus carrying DCX-ChR2-EGFP gene. After 48 hours of lentivirus infection, 470 nm blue laser irradiation was performed for 3 consecutive days. NeuN positive cell density(the maturation of neural stem cells)and the ratio of NeuN/Hoechst in each group were observed. Western blot was used to detect the expression levels of MAP2, NeuN, Neurog2, NeuroD1 and GluR2. Western blot was used to detect the expressions of β-catenin and TCF4 associated with Wnt/β-catenin signaling channel. Verapamil (100 μmol/L, L-type calcium channel blockers) and Dkk1 (50 μg/ml, β-catenin inhibitor) were used to treat stem cells of the NSCs+ChR2 group and then the expressions of MAP2, NeuN, Neurog2, NeuroD1 and GluR were detected by Western blot. RESULTS: After 3 days of 470 nm blue laser irradiation, NeuN positive cell density(the maturation of neural stem cells)and the ratio of NeuN/Hoechst, the expression levels of the protein MAP2, NeuN, Neurog2, NeuroD1, GluR and the protein β-catenin and TCF4 associated with Wnt/β-catenin signaling channel detected by Western blot were significantly increased in the group of NSCs+ChR2, compared with NSCs and NSCs+EGFP groups. The expressions of MAP2, NeuN, Neurog2, NeuroD1 and GluR were remarkably decreased after treated by verapamil and Dkk1 in the group of NSCs+ChR2. It was proved that the opening of ChR2 channel producing cationic influx promoted the maturation of neural stem cells and induced by the Wnt/β-catenin signaling pathway. CONCLUSION Optical genetic promoted the maturation of newborn neurons through the Wnt/β-catenin signaling pathway.
Animals ; Cells, Cultured ; Neural Stem Cells ; cytology ; Neurons ; cytology ; Optogenetics ; Rats ; Transfection ; Wnt Signaling Pathway

OBJECTIVE: To analyze the changes of blood biochemical index and the pathological changes of myocardium and kidney in type 2 diabetic mouse at different time points, which can provide the basis for the selection of type 2 diabetic modeling time for later research. METHODS: After 6 weeks of feeding with high-fat diet, 24 healthy male ICR mice were injected with streptozocin (STZ, 30 mg/kg) intraperitoneally for 5 days to establish diabetic models. After 9 days, a random blood glucose ≥ 11.1 mmol / L was measured as diabetic mice. 4, 6 and 8 weeks after successfully preparing the diabetic mouse, 8 diabetic mice (a group)would be sacrificed each time. Then the biochemical and pathological conditions were analyzed: ① the indexes of heart and kidney were calculated. ②the serum levels of creatine kinase (CK), lactate dehydrogenase (LDH), creatinine (Cr) and blood urine nitrogen (BUN) were determined. ③ Histopathological changes of myocardium and renal tissues were observed by hematoxylin and eosin (HE) staining. Masson staining was used to observe the fibrosis of myocardium. PAS staining was adopted to observe the pathological changes of renal tissue. In addition, 8 ICR male mice were taken as the control group. RESULTS: At the 4, 6 and 8 week, cardiac organ coefficient, the values of LDH and CK were all increased compared with the control group. Cardiomyocyte hypertrophy and myocardial fibrosis could be observed. Renal organ coefficient, the values of Cr and BUN were increased. Glomerular hypertrophy, basement membrane thickening and atrophy could be perceived. CONCLUSION At the 6 week, related biochemical and pathological changes in diabetic mice were comparatively obvious and breeding time was relatively short. Thus, 6 weeks after the preparation of the diabetic mice would be the optimal time for type 2 diabetes mellitus modeling, proper for inventions of drugs and other research purposes including pathology, physiology, biochemistry, etc.
Animals ; Diabetes Mellitus, Experimental ; pathology ; Diabetes Mellitus, Type 2 ; pathology ; Disease Models, Animal ; Kidney ; pathology ; Male ; Mice ; Mice, Inbred ICR ; Streptozocin

OBJECTIVE: To screen for pathogenic variants in the coding regions of STK11 gene among Chinese patients with Peutz-Jeghers syndrome (PJS). METHODS: Peripheral blood samples were collected from 64 patients. The coding regions of the STK11 gene were detected by PCR and Sanger sequencing. RESULTS: Fourty-eight patients were found to harbor STK11 gene variants, which included 39 types of variants consisting of missense, nonsense, insertional, deletional and splice site variants. Among 64 PJS patients, the detection rate of point variants was 75.00% (48/64), of which missense variants accounted for 29.17% (14/48), nonsense variants accounted for 29.17%(14/48), insertion variants accounted for 2.08% (1/48), deletional variants accounted for 10.42% (5/48), and splice site variants accounted for 29.17% (14/48). The detection rates of sporadic cases and those with a family history were 71.8% (28/39) and 80.0% (20/25), respectively. Two variants (c.250A>T, c.580G>A) occurred in 3 PJS probands. Thirteen variants were unreported previously and were considered to be pathogenic. CONCLUSION The detection rate of variants among Chinese PJS patients is similar to that of other countries. A number of novel common variant sites were discovered, which enriched the spectrum of PJS-related variants.
Asian Continental Ancestry Group ; China ; DNA Mutational Analysis ; Humans ; Peutz-Jeghers Syndrome ; genetics ; Protein-Serine-Threonine Kinases ; genetics

The aim of this study was to explore the neuroprotective effect and mechanism of XingNaoJing injections (XNJ) on cerebral ischemia injury and blood-brain barrier (BBB) disruption. Middle cerebral artery occlusion (MCAO) method was applicated to establish the model of cerebral ischemia/reperfusion (I/R) injury in rats. BBB permeability after I/R injury was assessed with the leaking amount of Evans Blue and the expression of occludin and ZO-1. The expression of NOD-like receptor family, pyrin domain containing (NLRP3) was checked to explore the inhibition of inflammation by XNJ. The results showed that XNJ could significantly increase the survival percent, decrease the infarct area and ameliorate neurological deficits and brain damage after I/R injury. Leaking amount of Evans Blue was reduced by XNJ, and the expression of tight junction protein, occludin and ZO-1 was also up-regulated by XNJ, which showed a role of protection on BBB disruption. The expression of NLRP3 was inhibited after exposure of XNJ, which was associated with inhibition of the inflammatory response. In summary, XNJ could suppress NLRP3 inflammasomes and improve BBB disruption and brain damage in rats after cerebral I/R injury, which provided a beneficial insight to further explore XNJ.

Resistance to cisplatin (DDP)-based chemotherapy is a major cause of treatment failure in human gastric cancer (GC). It is necessary to identify the drugs to re-sensitize GC cells to DDP. In our previous research, Zuo Jin Wan Formula (ZJW) has been proved could increase the mitochondrial apoptosis via cofilin-1 in a immortalized cell line, SGC-7901/DDP. Due to the immortalized cells may still difficult highly recapitulate the important molecular events in vivo, primary GC cells model derived from clinical patient was constructed in the present study to further evaluate the effect of ZJW and the underlying molecular mechanism. Immunofluorescent staining was used to indentify primary cultured human GC cells. Western blotting was carried out to detect the protein expression. Cell Counting Kit-8 (CCK-8) was used to evaluate cell proliferation. Flow cytometry analysis was performed to assess cell apoptosis. ZJW inhibited proliferation and induced apoptosis in primary DDP-resistant GC cells. Notably, the apoptosis in GC cells was mediated by inducing cofilin-1 mitochondrial translocation, down-regulating Bcl-2 and up-regulating Bax expression. Surprisingly, the level of p-AKT protein was higher in DDP-resistant GC cells than that of the DDP-sensitive GC cells, and the activation of AKT could attenuate ZJW-induced sensitivity to DDP. These data revealed that ZJW can increase the chemosensitivity in DDP-resistant primary GC cells by inducing mitochondrial apoptosis and AKT inactivation. The combining chemotherapy with ZJW may be an effective therapeutic strategy for GC chemoresistance patients.
Adult ; Aged ; Aged, 80 and over ; Apoptosis ; drug effects ; Cell Proliferation ; drug effects ; Cisplatin ; pharmacology ; therapeutic use ; Cofilin 1 ; metabolism ; Drug Resistance, Neoplasm ; drug effects ; Drugs, Chinese Herbal ; pharmacology ; Female ; Humans ; Male ; Middle Aged ; Mitochondria ; drug effects ; metabolism ; pathology ; Proto-Oncogene Proteins c-akt ; metabolism ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Stomach Neoplasms ; drug therapy ; metabolism ; pathology ; Tumor Cells, Cultured