[Acupressure with activating blood and dissipating blood stasis prescription for early swelling of calcaneal fractures].

OBJECTIVE: To observe clinical effects of acupressure combined with activating blood and dissipating blood stasis prescription for the treatment of early swelling of calcaneal fractures. METHODS: From June 2016 to April 2018, 78 patients with calcaneus were divided into observation group and control group. There were 39 patients in observation group, including 36 males and 3 females aged from 28 to 61 years old with an average of(41.7±13.3) years old; 20 patients were caused by falling down, 10 patients were caused by traffic accident and 9 patients were caused by other injuries; The time from injury to medical treatment ranged from 0.5 to 10.5 h with an average of(4.6±3.2) h; 15 patients were type II, 17 patients were type III and 7 patients were type IV according to Sanders classification; the patients began to be pressed the three points of "Xuehai(SP 10)" "Sanyinjiao(SP 6)" "Zusanli(ST 36)" for 10 min from the day of hospital admission to the day of operation, 3 times a day, and taken prescription of activating blood and dissipating blood stasis prescription orally for 5 days, one dose per day and twice a day. There were 39 patients in control group, including 34 males and 5 females aged from 26 to 62 years old with an average of (43.3±12.3) years old; 19 patients were caused by falling down, 12 patients were caused by traffic accident and 8 patients were caused by other injuries; the time from injury to medical treatment ranged from 1 to 11 h with an average of (4.4±3.7) h; 15 patients were typeII, 18 patients were type III and 6 patients were type IV according to Sanders classification; patients were given 20% mannitol intravenously for 5 days, twice a day, 125 ml each time from hospital admission. During the 5 days of admission, the degree of swelling of the affected ankle was measured and calculated by water spillover method every day. The result which was measured and calculated on the first day of admission without treatment was the degree of swelling before treatment, compared degree of swelling before operation and 1st, 2nd, 3rd and 4th day after treatment; the time needed for the first positive skin fold sign in the affected ankle and the number of cases of tension blisters between two groups before operation were observed. RESULTS: The swelling of the affected ankle on the1st, 2nd, 3rd and 4th day after treatment in the observation group separately were(12.67±0.82)%, (11.87±0.88)%, (10.65±0.92)%, (9.47±0.96)%, and lower than control group which separately were(13.31±0.98)%, (13.51±0.84)%, (12.22±0.94)%, (11.38±1.01)%; The time for the first appearance of the skin fold sign was (6.41±1.74) days in the observation group was earlier than that of the control group (8.15±2.01) days; There was no significant difference in occurrence of tension blisters between observation group(3 patients) and control group(9 patients). CONCLUSIONS: Acupressure combined with activating blood and dissipating blood stasis prescription for the treatment of early swelling of calcaneal fractures could accelerate the decline of swelling of the affected ankle, shorten the time of the first skin fold sign, reduce the incidence of tension blisters in the ankle, shorten the waiting time of operation, and the surgical treatment as soon as possible.

Brain networks modeling for studying the mechanism underlying the development of Alzheimer's disease.

Alzheimer's disease is a primary age-related neurodegenerative disorder that can result in impaired cognitive and memory functions. Although connections between changes in brain networks of Alzheimer's disease patients have been established, the mechanisms that drive these alterations remain incompletely understood. This study, which was conducted in 2018 at Northeastern University in China, included data from 97 participants of the Alzheimer's Disease Neuroimaging Initiative (ADNI) dataset covering genetics, imaging, and clinical data. All participants were divided into two groups: normal control (n = 52; 20 males and 32 females; mean age 73.90 ± 4.72 years) and Alzheimer's disease (n = 45, 23 males and 22 females; mean age 74.85 ± 5.66). To uncover the wiring mechanisms that shaped changes in the topology of human brain networks of Alzheimer's disease patients, we proposed a local naïve Bayes brain network model based on graph theory. Our results showed that the proposed model provided an excellent fit to observe networks in all properties examined, including clustering coefficient, modularity, characteristic path length, network efficiency, betweenness, and degree distribution compared with empirical methods. This proposed model simulated the wiring changes in human brain networks between controls and Alzheimer's disease patients. Our results demonstrate its utility in understanding relationships between brain tissue structure and cognitive or behavioral functions. The ADNI was performed in accordance with the Good Clinical Practice guidelines, US 21CFR Part 50-Protection of Human Subjects, and Part 56-Institutional Review Boards (IRBs)/Research Good Clinical Practice guidelines Institutional Review Boards (IRBs)/Research Ethics Boards (REBs).

Laparoscopic gastrectomy versus open gastrectomy for elderly patients with gastric cancer: a systematic review and meta-analysis.

BACKGROUND: The objective of this study was to evaluate the feasibility, safety, and potential benefits of laparoscopic gastrectomy (LG) comparing with open gastrectomy (OG) in elderly population. METHODS: Studies comparing LG with OG for elderly population with gastric cancer, published between January 1994 and July 2015, were identified in the PubMed, Embase, and ISI Web of Science databases. Operative outcomes (intraoperative blood loss, operative time, and the number of lymph nodes harvested) and postoperative outcomes (time to first ambulation, time to first flatus, time to first oral intake, postoperative hospital stay, postoperative morbidity) were included and analyzed. The Newcastle-Ottawa Scale was used to assess the quality of the pooled study. A funnel plot was used to evaluate the publication bias. RESULTS: Seven studies totaling 845 patients were included in the meta-analysis. LG in comparison to OG showed less intraoperative blood loss (weighted mean difference (WMD) -127.47; 95% confidence interval (CI) -202.79 to -52.16; P < 0.01), earlier time to first ambulation (WMD -2.07; 95% CI -2.84 to -1.30; P < 0.01), first flatus (WMD -1.04; 95% CI -1.45 to -0.63; P < 0.01), and oral intake (WMD -0.94; 95% CI -1.11 to -0.77; P < 0.01), postoperative hospital stay (WMD -5.26; 95% CI -7.58 to -2.93; P < 0.01), lower overall postoperative complication rate (odd ratio (OR) 0.39; 95% CI 0.28 to 0.55; P < 0.01), less surgical complications (OR 0.47; 95% CI 0.32 to 0.69; P < 0.01), medical complication (OR 0.35; 95% CI 0.22 to 0.56; P < 0.01), incisional complication (OR 0.40; 95% CI 0.19 to 0.85; P = 0.02), and pulmonary infection (OR 0.49; 95% CI 0.26 to 0.93; P = 0.03). No significant differences were observed between LG and OG for the number of harvested lymph nodes. However, LG had longer operative times (WMD 15.73; 95% CI 6.23 to 25.23; P < 0.01). CONCLUSIONS: LG is a feasible and safe approach for elderly patients with gastric cancer. Compared with OG, LG has less blood loss, faster postoperative recovery, and reduced postoperative morbidity.

A deficiency in chloroplastic ferredoxin 2 facilitates effective photosynthetic capacity during long-term high light acclimation in Arabidopsis thaliana.

Photosynthetic electron transport is the major energy source for cellular metabolism in plants, and also has the potential to generate excess reactive oxygen species that cause irreversible damage to photosynthetic apparatus under adverse conditions. Ferredoxins (Fds), as the electron-distributing hub in the chloroplast, contribute to redox regulation and antioxidant defense. However, the steady-state levels of photosynthetic Fd decrease in plants when they are exposed to environmental stress conditions. To understand the effect of Fd down-regulation on plant growth, we characterized Arabidopsis thaliana plants lacking Fd2 (Fd2-KO) under long-term high light (HL) conditions. Unexpectedly, Fd2-KO plants exhibited efficient photosynthetic capacity and stable thylakoid protein complexes. At the transcriptional level, photoprotection-related genes were up-regulated more in the mutant plants, suggesting that knockout Fd2 lines possess a relatively effective photo-acclimatory responses involving enhanced plastid redox signaling. In contrast to the physiological characterization of Fd2-KO under short-term HL, the plastoquinone pool returned to a relatively balanced redox state via elevated PGR5-dependent cyclic electron flow during extended HL. fd2 pgr5 double mutant plants displayed severely impaired photosynthetic capacity under HL treatment, further supporting a role for PGR5 in adaptation to HL in the Fd2-KO plants. These results suggest potential benefits of reducing Fd levels in plants grown under long-term HL conditions.

[Protective effect of Pinus massoniana bark extracts against cisplatin-induced nephrotoxicity in human embryonic kidney cells].

OBJECTIVE: To investigate the in vitro protective effect of Pinus massoniana bark extracts (PMBE) against cisplatin-induced nephrotoxicity in human embryonic kidney cells (HEK293), and preliminarily study its mechanism. METHOD: Human embryonic kidney cells (HEK293) were cultured in vitro. The MTT assay was adopted to test the effect of PMBE and cisplatin on growth of HEK293 cells, and the protective effect of PMBE on cisplatin-induced nephrotoxicity of HEK293, and then detect the intracellular reactive oxygen species (ROS), malondialdehyde (MDA), glutathione (GSH) content, catalase (CAT), superoxide dismutase (SOD) and activity of thioredoxin reductase (TrxR). RESULT: PMBE could promote growth of HEK293 cells at low concentrations, but generate slight nephrotoxicity at high concentration. Cisplatin could inhibit growth of HEK293 cells, increase ROS and MDA content, while reducing SOD, CAT and TrxR. The pre-protective PMBE was added to reduce cisplatin's injury to HEK293 cells, ROS, MDA and GSH content, SOD, CAT and TrxR within certain range. CONCLUSION: PMBE at specific concentration has the protective effect in cisplatin-induced nephrotoxicity in HEK293 cells. Its mechanism may be related to PMBE's antioxidant activity.

Antitumor effects of Pinus massoniana bark extract in murine sarcoma S180 both in vitro and in vivo.

Pinus massoniana bark extract (PMBE) is a mixture of flavonoids, and showed a capability of inducing cell apoptosis; however, its properties have not yet been fully investigated. This paper evaluates the antitumor effects of PMBE in murine sarcoma S180 both in vitro and in vivo. In vitro, the growth inhibition of S180 cells was concentration dependent on PMBE as shown by the CCK-8 assay. The AO/EB staining and flow cytometry assay showed that PMBE induced S180 cell apoptosis. Cell cycle analysis revealed that the cells in the S phase were decreased by treatment with PMBE. In vivo, the treatment of 100, 200, and 300 mg/kg PMBE reduced the tumor weight and volume of S180-bearing NIH mice by 9%-67% and 13%-68%, respectively. Peripheral leukocyte count and lymphoproliferation were increased significantly after treatment with PMBE. Our results suggest that PMBE inhibits the tumor cell growth by inducing cell apoptosis and improving lymphoproliferation.

[Case-control study on application of auricular acupuncture for the treatment of analgesia during perioperative period in total hip arthroplasty].

OBJECTIVE: To investigate clinical effects of auricular acupoint (AA) in the treatment of analgesia during perioperative period in total hip arthroplasty. METHODS: From March 2008 to August 2010, 60 patients with late osteonecrosis of the femoral head were treated by total hip arthroplasty and randomly divided into auricular acupuncture (AA) group and control group, 30 patients in each group. There were 11 males and 19 females in the AA group,with an average age of (60.93 +/- 5.90) years; the patients were treated with auricular acupuncture on the point of Shenmen, Subcortex, Kidney and hip joint for four times a week. There were 12 males and 18 females in control group, with an average age of (59.87 +/- 6.21) years; while the patients without auricular acupuncture. VAS score was used to evaluat the degree of pain; Harris score was used to evaluat the function of hip joint. All patients received patient controlled analgesia pump (PCA) for 48 hours after surgery (400 ml liquids were in PCA pump, including 800 mg tramadol and 0.8 mg fentanyl). The dosage of liquids and adverse reaction of PCA pump in different time were recorded. RESULTS: The VAS score on the 3rd, 4th, 5th and 7th day separately was (3.61 +/- 0.29), (3.59 +/- 0.30), (2.97 +/- 0.26), (2.29 +/- 0.45), and lower than control group, which separately was (4.19 +/- 0.28), (4.00 +/- 0.31), (3.15 +/- 0.29), (2.83 +/- 0.31). The dosage of PCA in AA group separately was (72.27 +/- 8.06), (60.40 +/- 8.16), (44.13 +/- 4.75), (40.40 +/- 3.69), and less than control group, which was (86.27 +/- 8.51), (73.87 +/- 8.32), (54.53 +/- 5.20), (44.67 +/- 6.31) on the time of 0-12, 12-24, 24-36 h and 36-48 h after surgery. During the using of PCA, nausea and vomiting occurred in 5 cases, less than control group (21 cases). Harris score in AA group (78.90 +/- 5.14) was higher than control group (73.37 +/- 5.99) 2 weeks after operation. CONCLUSION: Auricular acupuncture can reduce postoperative pain, reduce the usage of analgesic and complications, such as nausea and vomiting, improve the function of hip joint after operation.

[Experimental study on Yougui recipe in preventing osteolysis surrounding artificial prosthesis].

OBJECTIVE: To explore effects of Yougui recipe (see text) and salmon calcitonin acetate in preventing osteolysis surrounding artificial prosthesis. METHODS: Thirty-two SD male rats with weighted (250 +/- 20) g, aged 8 weeks, were randomly divided into four groups: blank group, model group, salmon calcitonin acetate group and Yougui recipe (see text) group, and 8 rats in each group. Blank group did not undergo any process, other 24 rats underwent anesthesia by chloral hydrate, their knee joints were exposed through medial patellar side,drilling from fermoral condyle nest to marrow cavity,high density of polythlene particles were injected into hole, titanium nail were put into, bone wax closed the window, then suturing step by step. After the molding, saline were used to gavaged in blank group and model group, Yougui recipe (see text) for Yougui recipe (see text) group, salmon calcitonin maximus injection for calcitonin group. After 10 weeks' mediation, rats were executed, and arterial blood and bilateral femoral organization were collected to biochemical, imaging morphology, tissue pathology and molecular biology detection. RESULTS: The key gene expression of activiting osteoclast were inhibited in Yougui recipe (see text) group and calcitonin group. The level of OPG, Ca, ALP in Yougui recipe group were higher than calcitonin group (P<0.01); the content of RANKL were lower (P<0.01). There were no significance meaning in RANK, Trap5b, P between two groups. CONCLUSION: Both of Yougui recipe (see text) and calcitonin can slow and treat surrounding osteolysis of artificial joint prosthesis, and Yougui recipe (see text) has better effect in promoting bone formation. The effect of Yougui recipe (see text) in promoting bone formation, inhibiting osteoclasts to provide a new method to treating surrounding osteolysis of artificial joint prosthesis.

MpAsr encodes an intrinsically unstructured protein and enhances osmotic tolerance in transgenic Arabidopsis.

Abscisic acid-, stress- and ripening (ASR) -induced proteins are plant-specific proteins whose expression is up-regulated under abiotic stresses or during fruit ripening. In this study, we characterized an ASR protein from plantain to explore its physiological roles under osmotic stress. The expression pattern of MpAsr gene shows that MpAsr gene changed little at the mRNA level, while the MpASR protein accumulates under osmotic treatment. Through bioinformatic-based predictions, circular dichroism spectrometry, and proteolysis and heat-stability assays, we determined that the MpASR protein is an intrinsically unstructured protein in solution. We demonstrated that the hydrophilic MpASR protein could protect L: -lactate dehydrogenase (L: -LDH) from cold-induced aggregation. Furthermore, heterologous expression of MpAsr in Escherichia coli and Arabidopsis enhanced the tolerance of transformants to osmotic stress. Transgenic 35S::MpAsr Arabidopsis seeds had a higher germination frequency than wild-type seeds under unfavorable conditions. At the physiological level, 35S::MpAsr Arabidopsis showed increased soluble sugars and decreased cell membrane damage under osmotic stress. Thus, our results suggest that the MpASR protein may act as an osmoprotectant and water-retaining molecule to help cell adjustment to water deficit caused by osmotic stress.

Pinus massoniana bark extract protects against oxidative damage in L-02 hepatic cells and mice.

The hepatoprotective activity of Pinus massoniana bark extract (PMBE) against hydrogen peroxide (H2O2)-induced damage in normal human liver L-02 cells and carbon tetrachloride (CCl4)-induced acute hepatotoxicity in mice was investigated. The L-02 cells were pre-treated with PMBE for 24 hours prior to exposure to 0.5 mM H2O2 for 3 or 24 hours. The cell viability, level of malondialdehyde (MDA) and glutathione (GSH), and the catalase (CAT) activity were evaluated. For in vivo experiments, mice were divided into groups and PMBE administered orally, after which each group was assigned a further treatment. Histopathological examination, the serum levels of alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), and GSH, the liver tissue levels of MDA and GSH, the activities of CAT and glutathione peroxidase (GSH-Px), were evaluated. PMBE treatment decreased the level of MDA and increased the cell viability, GSH content and CAT activity in H2O2 treated L-02 cells treated for 3 hours. PMBE obviously decreased serum ALT, AST, ALP, and liver tissue MDA, while increasing serum GSH, and liver tissue CAT and GSH-Px activities. In conclusion, PMBE treatment prevents H2O2 and CCl4-induced liver damage, and therefore could have a potential clinical usage.

Characterization of a novel plantain Asr gene, MpAsr, that is regulated in response to infection of Fusarium oxysporum f. sp. cubense and abiotic stresses.

Asr (abscisic acid, stress, ripening induced) genes are typically upregulated by a wide range of factors, including drought, cold, salt, abscisic acid (ABA) and injury; in addition to plant responses to developmental and environmental signals. We isolated an Asr gene, MpAsr, from a suppression subtractive hybridization (SSH) cDNA library of cold induced plantain (Musa paradisiaca) leaves. MpAsr expression was upregulated in Fusarium oxysporum f. sp. cubense infected plantain leaves, peels and roots, suggesting that MpAsr plays a role in plantain pathogen response. In addition, a 581-bp putative promoter region of MpAsr was isolated via genome walking and cis-elements involved in abiotic stress and pathogen-related responses were detected in this same region. Furthermore, the MpAsr promoter demonstrated positive activity and inducibility in tobacco under F. oxysporum f. sp. cubense infection and ABA, cold, dehydration and high salt concentration treatments. Interestingly, transgenic Arabidopsis plants overexpressing MpAsr exhibited higher drought tolerance, but showed no significant decreased sensitivity to F. oxysporum f. sp. cubense. These results suggest that MpAsr might be involved in plant responses to both abiotic stress and pathogen attack.

Carrot antifreeze protein does not exhibit the polygalacturonase-inhibiting activity of PGIP family.

The carrot (Daucus carota) antifreeze protein (DcAFP) has a strong antifreeze activity and identified as belonging to the plant polygalacturonase-inhibiting protein (PGIP) family based on its sequence similarities, including the presence of a leucine-rich repeat (LRR) motif. In this study, yeast two-hybrid technology was used to analyze whether the carrot AFP could act as a PGIP. The complete DcAFP and polygalacturonase (PGase; obtained from fungus Alternaria alternata by RT-PCR) coding sequences were cloned into the bait and capture vectors, respectively, and yeast two-hybrid assays were performed. The results revealed that there was no evidence of an interaction between DcAFP and PGase, which suggests that DcAFP probably lacks PGIP activity. An analysis of the electrostatic potential of DcAFP and other PGIPs revealed that a large number of nonconservative residues within the beta-helix of the DcAFP LRR motif had been substituted to basic amino acids, thus changing the surface from negative to positive. This will electrostatically prevent DcAFP from binding with the positively charged surface of PGase. This is the first report that showed the correlation between nonconservative amino acids within the LRR motif of the DcAFP and its loss of polygalacturonase inhibiting activity.

Effects of Pinus massoniana bark extract on cell proliferation and apoptosis of human hepatoma BEL-7402 cells.

AIM: To study the effects of Pinus massoniana bark extract (PMBE) on cell proliferation and apoptosis of human hepatoma BEL-7402 cells and to elucidate its molecular mechanism. METHODS: BEL-7402 cells were incubated with various concentrations (20-200 microg/mL) of PMBE for different periods of time. After 48 h, cell proliferation was determined by 3-(4,5-dimethyl-thiazolyl-2)-2,5-diphenyl tetrazolium bromide (MTT) assay. Apoptosis was evaluated by morphological observation, agarose gel electrophoresis, and flow cytometry analysis. Possible molecular mechanisms were primarily explored through immunohistochemical staining. RESULTS: PMBE (20-200 microg/mL) significantly suppressed BEL-7402 cell proliferation in a time- and dose-dependent manner. After treatment of BEL-7402 cells with 160 microg/mL PMBE for 24, 48, or 72 h, a typical apoptotic "DNA ladder" was observed using agarose gel electrophoresis. Nuclear condensation and boundary aggregation or split, apoptotic bodies were seen by fluorescence and electron microscopy. Sub-G1 curves were displayed by flow cytometry analysis. PMBE decreased the expression levels of Bcl-2 protein in a time-dependent manner after treatment of cells with 160 microg/mL PMBE. CONCLUSION: PMBE suppresses proliferation of BEL-7402 cells in a time- and dose-dependent manner and induces cell apoptosis by possibly downregulating the expression of the bcl-2 gene.

Expression, purification, and antifreeze activity of carrot antifreeze protein and its mutants.

Antifreeze proteins (AFPs) enable organisms to survive under freezing or sub-freezing conditions. AFPs have a great potential in the low temperature storage of cells, tissues, organs, and foods. This process will require a large number of recombinant AFPs. In the present study, the recombinant carrot AFP was highly expressed in Escherichia coli strain BL21 (DE3). The activity of the purified and refolded recombinant proteins was analyzed by measurement of thermal hysteresis (TH) activity and detection of in vitro antifreeze activity by measuring enhanced cold resistance of bacteria. Two carrot AFP mutants generated by site-directed mutagenesis were also expressed and purified under these conditions for use in parallel experiments. Recombinant DcAFP displayed a TH activity equivalent to that of native DcAFP, while mutants DcAFP-N130Q and rDcAFP-N130V showed 32 and 43% decreases in TH activity, respectively. Both the recombinant DcAFP and its mutants were able to enhance the cold resistance of bacteria, to degrees consistent with their respective TH activities.

Significance of conservative asparagine residues in the thermal hysteresis activity of carrot antifreeze protein.

The approximately 24-amino-acid leucine-rich tandem repeat motif (PXXXXXLXXLXXLXLSXNXLXGXI) of carrot antifreeze protein comprises most of the processed protein and should contribute at least partly to the ice-binding site. Structural predictions using publicly available online sources indicated that the theoretical three-dimensional model of this plant protein includes a 10-loop beta-helix containing the approximately 24-amino-acid tandem repeat. This theoretical model indicated that conservative asparagine residues create putative ice-binding sites with surface complementarity to the 1010 prism plane of ice. We used site-specific mutagenesis to test the importance of these residues, and observed a distinct loss of thermal hysteresis activity when conservative asparagines were replaced with valine or glutamine, whereas a large increase in thermal hysteresis was observed when phenylalanine or threonine residues were replaced with asparagine, putatively resulting in the formation of an ice-binding site. These results confirmed that the ice-binding site of carrot antifreeze protein consists of conservative asparagine residues in each beta-loop. We also found that its thermal hysteresis activity is directly correlated with the length of its asparagine-rich binding site, and hence with the size of its ice-binding face.