Electroacupuncture may alleviate diabetic neuropathic pain by inhibiting the microglia P2X4R and neuroinflammation.

Diabetic neuropathic pain (DNP) is a common and destructive complication of diabetes mellitus. The discovery of effective therapeutic methods for DNP is vitally imperative because of the lack of effective treatments. Although 2 Hz electroacupuncture (EA) was a successful approach for relieving DNP, the mechanism underlying the effect of EA on DNP is still poorly understood. Here, we established a rat model of DNP that was induced by streptozotocin (STZ) injection. P2X4R was upregulated in the spinal cord after STZ-injection. The upregulation of P2X4R was mainly expressed on activated microglia. Intrathecal injection of a P2X4R antagonist or microglia inhibitor attenuated STZ-induced nociceptive thermal hyperalgesia and reduced the overexpression of brain-derived neurotrophic factor (BDNF), interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) in the spinal cord. We also assessed the effects of EA treatment on the pain hypersensitivities of DNP rats, and further investigated the possible mechanism underlying the analgesic effect of EA. EA relieved the hyperalgesia of DNP. In terms of mechanism, EA reduced the upregulation of P2X4R on activated microglia and decreased BDNF, IL-1ß and TNF-α in the spinal cord. Mechanistic research of EA's analgesic impact would be beneficial in ensuring its prospective therapeutic effect on DNP as well as in extending EA's applicability.

Development of FAP-Targeted Chimeric Antigen Receptor NK-92 Cells for Non-Small Cell Lung Cancer.

OBJECTIVES: Over the past two decades, great progress has been made in advancing the early detection and multimodal treatment of non-small cell lung cancer (NSCLC). However, overall cure rates and survival rates of NSCLC are still not satisfactory, and research into new therapies is needed. This study attempted to construct human Fibroblast Activation Protein-Chimeric Antigen Receptor Natural killer (NK)-92 cells (hFAP-CAR-NK-92 cells) and explore their potential therapeutic effects in NSCLC. METHODS: Immunohistochemistry analysis was carried out to examine fibroblast activation protein (FAP) and Gasdermin E (GSDME) expression in clinical specimens of lung adenocarcinoma and squamous cell carcinoma tissue. Then the engineered hFAP-CAR-NK-92 cells efficiency was determined in vitro with lactate dehydrogenase (LDH) cytotoxicity assay and the cell morphology of A549, H226, and cancer-related fibroblast (CAF) was observed by electron microscopy. After the co-culture of target cells and effect cells, flow cytometry was employed for examining the CD107a expression in the effect cells, and western blotting was conducted for the cleavage levels of Caspase 3 and GSDME proteins in the target cells. The safety and efficacy of hFAP-CAR-NK-92 cells adoptive transfer immunotherapy in a tumor-bearing mouse were evaluated. RESULTS: Clinical studies have shown FAP positivity in patients with NSCLC. Compared with A549 or H226 cells alone, FAP expression was notably raised in A549+CAF cells or H226+CAF cells in nude mice, respectively (p < 0.05). The killing efficiency of K562 cells was not significantly different between hFAP-CAR-NK-92 and NK-92 cells (p > 0.05). The hFAP-CAR-NK-92 cells presented a higher killing efficiency against the hFAP-target (A549-hFAP, H226-hFAP and CAF-hFAP) cells than the NK-92 cells (p < 0.05). The degranulation of CD107a and cleavage levels of GSDME and Caspase 3 protein in the hFAP-CAR-NK-92 group were higher than those in the NK-92 group (p < 0.05). The 300 nM Granzyme B also induced pyroptosis in hFAP- or GSDME-positive cells (p < 0.05). In vivo experiments revealed that hFAP-CAR-NK-92 cells inhibited tumor progression of hFAP-positive NSCLC (p < 0.05). CONCLUSIONS: In this study, we successfully constructed hFAP-CAR-NK-92 cells and confirmed that hFAP-CAR-NK-92 cells could target hFAP-positive NSCLC to inhibit the progression of NSCLC by activating the Caspase-3/GSDME pyroptosis pathway.

[Risk Assessment Method of Non-point Source Pollution Output for Watershed Using High Resolution Data].

In recent years, non-point source pollution has become the main cause of water quality deterioration in some reservoirs in China. Taking the Panjiakou Reservoir as an example, the classical output risk model was improved by introducing a precipitation factor and terrain factor. Combined with high-resolution satellite precipitation products (GPM) and GF-6 satellite images, a high-resolution data-driven risk assessment method for non-point source pollution output was established to study the temporal and spatial distribution characteristics of non-point source pollution output risk in the Panjiakou Reservoir basin. The results showed that the non-point source pollution output risk was high in the study area in 2018. The areas with higher and highest risk of nitrogen pollution output accounted for approximately 70.6% of the total watershed area, whereas the higher risk of phosphorus pollution output accounted for approximately 21.9%. The temporal and spatial distribution characteristics of non-point source pollution output risk in the Panjiakou Reservoir basin were analyzed. It was found that the non-point source pollution output risk in the Panjiakou Reservoir basin increased first and then decreased from April to September. This was consistent with the spatial and temporal distribution of precipitation in the basin. Combined with the analysis of land use distribution characteristics, the upstream area of the basin was mainly cultivated land, whereas cities were concentrated in the downstream portion of the basin. Affected by agricultural production and human activities, the risk of non-point source pollution output was higher in these regions. In view of the temporal and spatial distribution characteristics of non-point source pollution output risk, it is necessary to formulate a reasonable agricultural fertilization method, plan the landscape layout of source-sinks, and construct vegetation buffer zones.

A programmable and sensitive CRISPR/Cas12a-based MicroRNA detection platform combined with hybridization chain reaction.

MicroRNAs (miRNAs) play an essential role in cancer diagnosis and prognosis. Developing a new method for sensitive detection of miRNA is constantly in demand. CRISPR/Cas12a system can nonspecifically cleave single-stranded DNA after specific recognition of target DNA, showing tremendous potential in molecular diagnostics. However, CRISPR-based detection methods require synthesizing different crRNAs for detecting different targets, which limit their widespread application. Herein, we design a versatile and sensitive miRNA detection platform based on CRISPR/Cas12a system combined with a hybridization chain reaction (HCR) circuit. In this design, the HCR circuit as the signal transducer converts each miRNA into multiple DNA duplexes, which act as the activators to activate the trans-cleavage activity of Cas12a for further signal amplification. More importantly, this platform can sensitively detect different miRNAs without changing the spacer sequence of crRNA due to the fixed activators formed by HCR. In addition, the consistency between the proposed platform and RT-qPCR in miRNA detection extracted from different cell lines validated its practicability, demonstrating the potential in clinical diagnosis of cancers and monitoring therapy.

CD103-CD23+ classical hair cell leukemia: A case report and review of the literature.

INTRODUCTION: This case report is presented to improve our understanding of the atypical immunophenotype of hairy cell leukemia. PATIENT CONCERNS: A 58-year-old woman presented to our department with fatigue for >10 days. DIAGNOSIS: The patient was diagnosed with an increased proportion of abnormal lymphocytes in peripheral blood and bone marrow smear, positive for CD11c, CD19, CD20, CD22, CD25, CD123, CD200, and Kappa, partial expression of CD23, but no expression of CD103, positive for BRAF V600E mutation. INTERVENTIONS AND OUTCOMES: Cladribine combined with rituximab achieved complete remission of minor residual disease negativity. CONCLUSION: Hairy cell leukemia is rare, and the diagnosis and differential diagnosis should be made by combining the patient's medical history, clinical manifestations, immunophenotype, gene detection, and other means. Purine nucleoside analogs are the first-line treatments.

Effects of Peroral Endoscopic Myotomy on Esophageal Function in the Treatment of Achalasia.

Peroral endoscopic myotomy (POEM) is a new technique to treat achalasia, but the effects on esophageal motor function and structure are still unclear. This study aimed to examine the esophageal function and anatomical changes of patients with achalasia treated with POEM. This was a retrospective study of 43 patients with achalasia treated with POEM between January 2013 and January 2016 at the Second Affiliated Hospital of Xi'an Jiaotong University. The patients were grouped as previous treatments for achalasia (n = 19) versus no previous treatment (n = 24). Surgical success (defined as Eckardt score ≤3 points or decreased by >3 points compared with baseline), recurrence, and reintervention were analyzed. Three patients (7.0%) were Eckardt grade I, 16 (37.2%) were grade II, and 24 (55.8%) were grade III. Operation time was 35 to 150 (median = 49) minutes. Both groups showed improvements in the Eckardt score after surgery (both P < .001), without a difference between the 2 groups (P = .749). The maximal mean diameter of the esophagus was reduced, and the lower esophageal sphincter pressure was improved after surgery (both groups, all P < .001), without difference between the 2 groups (all P > .05). One case of failure was probably due to the presence of an esophageal stent. POEM has a high success rate and is possibly unaffected by previous treatments, except maybe stent implantation. Clinical symptoms of achalasia are significantly relieved by POEM; the function of the esophageal sphincter and the esophagus structure are improved. Previous esophageal stent implantation could increase failure likelihood, but this will have to be confirmed.

Transcutaneous Electrical Acupoint Stimulation Improves the Outcomes of In Vitro Fertilization: A Prospective, Randomized and Controlled Study.

OBJECTIVES: To explore whether transcutaneous electrical acupoint stimulation (TEAS) can improve the outcomes of in vitro fertilization (IVF). DESIGN: A prospective, randomized, and controlled study. SETTING: IVF center in a university hospital. PARTICIPANTS: Four hundred and eighty-one infertile patients with bilateral tubal blockage who were referred for IVF. Patients were randomized into four groups. INTERVENTION: TEAS was administered for 30min, respectively, at 24h before TVOR and two hours before ET. The acupoints included SP10 (Xuehai, bilateral), SP8 (Diji, bilateral), LR3 (Taichong, bilateral), ST36 (Zusanli, bilateral), EX-CA1 (Zigong, bilateral), RN4 (Guanyuan), PC6 (Neiguan, bilateral), and RN12 (Zhongwan). Based on different frequencies of TEAS, patients were grouped into a TEAS-2Hz group, a TEAS-100Hz group and a TEAS-2/100Hz group. Patients in the control group only received routine IVF treatment and no TEAS was applied on them. PRIMARY AND SECONDARY OUTCOME MEASURES: The number of mature oocytes, normally fertilized oocytes and good-quality embryos were used to evaluate oocyte developmental competence of the patients. Data of clinical pregnancy rate (CPR), implantation rate (IR), and live birth rate (LBR) were also obtained. The levels of neuropeptide Y (NPY), transforming growth factor alpha and granulocyte colony-stimulating factor in the follicular fluids were measured with enzyme-linked immunosorbent assay (ELISA). RESULTS: No significant differences were found between the control, TEAS-2Hz, TEAS-100Hz and TEAS-2/100Hz groups on the numbers of metaphase II oocytes, normally fertilized zygotes, early cleavage embryos or good quality embryos (P > .05). However, the CPR, IR and LBR of the TEAS-2/100Hz group were significantly higher than those of the other groups, respectively (P < .05). The NPY levels in the follicular fluids of TEAS-2/100Hz group were significantly higher than those of the other groups (P < .05). CONCLUSION: TEAS using a frequency of 2/100Hz could help to improve the IVF outcomes partly by increasing NPY levels in the follicular fluids.

[Effect of RAAS antagonist on the expression of gap junction cx43 in myocardium of spontaneously hypertensive rat].

UNLABELLED: OBJECTIVE To investigate the expression of gap junction protein Cx43 in the cardiac muscle of spontaneous hypertensive rat and the effects of various antagonists against renin angiotensin aldosterone system (RAAS) on Cx43 expression. METHODS 70 spontaneous hypertensive rats of 8-week age, 200-gram weight were separated into 7 groups, as hypertension, ramipril, telmisartan, eplerenone, ramipril + telmisartan, telmisartan + eplerenone, and ramipril+eplerenone treatment group. Another 10 healthy Wistar rats of the same age and weight were used as control group. All the rats were given intragastric administration at 8 a. m. every morning, and measured arteria caudilis pressure at 0, 4 and 8 week, respectively. 8 weeks later, all the rats were sacrificed, and the hearts were taken to measure the weight of left ventricle and the ratio of left ventricle to body weight. Myocardial fibrosis was observed by H&E staining of paraffin embedded sections, and Cx43 expression was examined by RT-PCR and western blot. RESULTS: The arteria caudilis pressure of spontaneous hypertensive rats was significantly higher than that of healthy control Wistar rats (P < 0.01). The decreased blood pressure was observed in RAAS antagonists treated rats, compared with hypertension group (P < 0.05). The combined treatment of telmisartan and eplerenone had the best effect of lowering blood pressure. Moreover, the weight of left ventricle, the ratio of left ventricle to body weight, myocardial fibrosis and angiotensin 11 were all prominently decreased in telmisartan and eplerenone combination group (P < 0.01). The expression of Cx43 in spontaneous hypertensive rats was significantly lower than that of healthy control Wistar rats (P < 0.01). Increased Cx43 expression was observed in RAAS antagonists treated rats, compared with hypertension group (P < 0.05). CONCLUSION: The expression of gap junction protein Cx43 was significantly down-regulated in spontaneous hypertensive rats, while RAAS antagonists increased Cx43 expression. The combination of telmisartan and eplerenone effectively recovered the expression of Cx43 and probably reversed hypertension.

[A method for screening microRNA target genes in goat skin by microRNA seed sequence mediated PCR].

The purpose of the present study was to establish a new microRNA seed mediated controllable genetic operation, namely MicroRNA Targets Finger Print (MTFP), for screening microRNA targets and detecting target gene expression profiles. Based on combining the complementary sequence of seed sequence, upstream and downstream anchor sequence including special adaptor, microRNA targets were amplified by means of the reverse transcription and special two step PCR. The polyacrylamide gel electrophoresis was used to analyze the sizes of amplified microRNA targets and their abundance of expression, which were used to screen specifically expressed genes in different physiological or experimental conditions. Specific target genes were obtained through isolation of DNA fragments and sequencing methods. As an example, by screening the targets of miR-203 in goat skin, five genes were amplified and sequenced with the sizes of 718 bp (JN709494), 349 bp (JN709495), 243 bp (JN709496), 159 bp (JN709497), and 97 bp (JN709498) from goat skin collections. The novel universal MTFP method could be applied for finding microRNA regulation targets or assessing target gene expression profile.

3,5-Dicaffeoylquinic acid isolated from Artemisia argyi and its ester derivatives exert anti-leucyl-tRNA synthetase of Giardia lamblia (GlLeuRS) and potential anti-giardial effects.

An aqueous ethanol extract of Artemisia argyi inhibited the aminoacylation activity of LeuRS from Giardia lamblia (GlLeuRS). The bioassay-guided fractionation of the extract led to the isolation of 3,5-dicaffeoylquinic acid (1), with an IC50 of 5.82 µg/mL. The ester derivatives of 1 were also found to possess strong anti-GlLeuRS effects, with IC50 values of 1.79, 5.51 and 2.56 µg/mL respectively. Anti-giardial assay showed that the derivatives, especially 3,5-dicaffeoylquinic acid propyl ester (4) (IC50=4.62 µg/mL), were effective at killing G. lamblia.

[Effects of conservation tillage on soil water conservation and crop yield of winter wheat-spring maize rotation field in Weibei highland].

A field experiment was conducted in 2007-2010 to study the effects of no-tillage, subsoiling, and deep-ploughing combined with balanced fertilization, traditional fertilization, and no (or lower amount) fertilization on the soil water storage, crop yield, water use efficiency (WUE), and economic return of winter wheat-spring maize rotation field in Weibei highland. Among the tillage measures, no-tillage in fallow period had the best effect in soil water conservation, followed by sub-soiling, and deep-ploughing. The average water storage in 0-200 cm soil layer in crop growth period under no-tillage and sub-soiling was 6.7% and 1.9% higher than that under deep-ploughing, respectively. Under the balanced, traditional, and no (or lower amount) fertilizations, subsoiling all showed the highest yield, WUE, and economic return, with the best effect under balanced fertilization. The three-year crop yield under sub-soiling combined with balanced fertilization was 6909, 9689, and 5589 kg x hm(-2), WUE was 18.5, 25.2, and 23.0 kg x hm(-2) x mm(-1), and economic return was 5034, 5045, and 7098 yuan x hm(-2), respectively. It was suggested that balanced fertilization combined with sub-soiling had the best effect in soil water conservation and yield- and income increase, being the more appropriate fertilization and tillage mode for the wheat-maize rotation field in Weibei highland.

[Study on the microstructure and ESR dosimetry of tetracycline-stained teeth with SEM observation and ESR measurement].

PURPOSE: To compare the microstructure and ESR dosimetry between tetracycline-stained teeth and normal teeth by means of scanning electron microscopy (SEM) and electron spin resonance (ESR) dosimeter. METHODS: Ten first or second premolars of tetracycline-stained teeth and ten normal teeth extracted for adult orthodontic persons were collected. The enamel on the surface and the dentine on the cross section of both type of teeth were observed with SEM. The ESR signal of teeth components (enamel and dentine) was evaluated by X-band ESR spectroscopy. RESULTS: Compared with normal teeth, the enamel of tetracycline-stained teeth was of porosity and the enamel prisms were irregular. The dentinal tubules and dentinal matrix also showed obvious difference between the two type of teeth. The X-band ESR spectrum of tetracycline-stained teeth was different from normal teeth. CONCLUSION: The microstructure and the native radicals have significant effect on the tetracyclines deposited in the teeth.

[Randomized controlled clinical trials for electroacupuncture treatment of urinary incontinence in stroke patients].

OBJECTIVE: To observe clinical efficacy of electroacupuncture (EA) for post-stroke urinary incontinence in patients. METHODS: A total of 111 stroke inpatients with urinary incontinence were randomly divided into EA group (n = 56) and control group (n = 55). Patients of the control group were treated by administration of calcium ion antagon, angiotensin-converting enzyme inhibitor, angiotensin II receptor antagonist, compound thromb-clearing agent (i.v.), etc.,and acupuncture of Jianyu (LI 15), Xuehai (SP 10), etc. Patients of the EA group were treated by EA of bilateral Shenshu (BL 23), Huiyang (BL 35), etc., combined with the treatment as the control group. The treatment was conducted once daily for 4 weeks. Crooks (1995) Scores of the clinical symptoms and clinical efficacy assessment were given before and after the treatment. RESULTS: After the treatment, the severity of urinary incontinence and symptom scores in both control and EA groups were decreased significantly in comparison with pre-treatment (P < 0.01). Following the treatment, of the 55 and 56 stroke patients in the control and EA groups, 0 and 4 (7.1%) were cured in the symptoms of frequent micturition, urgency of micturition and incontinence of urine, 20 (36.4%) and 35 (62.5%) were effective, 35 (63.6%) and 17 (30.4%) were invalid, with the effective rates being 36.4% and 69.6%, respectively. The therapeutic effect of EA group was apparently superior to that of the control group (P < 0.01). In addition, the infection rate of the EA group was strikingly lower than that of the control group (P < 0.05). CONCLUSION: EA can lower the severity of urinary incontinence and improve clinical symptoms of micturition in stroke patients.

[Correlation of human resistin gene expression with leukemia incidence].

Although the effect of mouse resistin on insulin-resistance has been well defined, but the biological function of human resistin is still unknown. This study was aimed to explore the possible physiological and pathological effects of human resistin, as well as the tissue distribution of human resistin and correlation of resistin gene expression with leukemia incidence. 152 leukemia patients without inflammatory complication and 100 healthy persons were selected as experimental and control groups respectively. The blood samples were collected, the total RNA was extracted, the expression distribution of resistin in different tissues was detected by semi-quantitative RT-PCR and then the statistical analysis was carried out. The results indicated that the expression of the human resistin gene was detected in normal fetus liver, adult bone marrow and umbilical cord blood and peripheral blood cells, while the resistin gene could not be amplified in fat, umbilical cord, placenta and adult liver. The resistin expression was detected in 21% leukemia patients and 27% healthy persons. The difference of the resistin gene expression between the two groups was not statistically significant (p>0.05). It is concluded that the higher expression of resistin exists in normal human fetus liver, adult bone marrow, umbilical cord blood and peripheral blood cells, which indicates that the distribution of human resistin correlates with normal hematopoiesis in certain extent, but its expression level and rate may not correlate with the incidence of leukemia.

[Immunoregulation of IL-18 in mice with radiated damage].

AIM: To explore the immunoregulation of IL-18 in the mice radiated by 60Co. METHODS: 32 C57 mice were radiated by 60Co and then treated by IL-18.2 weeks later, the transformation of lymphocytes, the ability of NK cells to kill tumor cells, the subtype of T cells and the content of IgG in serum were tested. RESULTS: IL-18 increased the function of lymphocyte transformation in 60Co radiated mice, enhanced the cytotoxic activity of NK cells against tumor cells of A375, U973 and KG1, and up-regulated the amount of CD4+T cells. However, the level of IgG in the serum of the radiated mice was not regulated by IL-18. CONCLUSION: IL-18 can enhance the immune function of the mice radiated by 60Co.

[Construction of apoptin gene delivery system and its effect on apoptosis of A375 cells].

AIM: To construct the delivery nanoparticles system of apoptin gene with O-carboxymethylated chitosan(CMC) and study its effect on inducing apoptosis of human melanoma cells A375 in vitro. METHODS: CMC nanoparticles containing apoptin gene were prepared by an ultrasonic method. Restriction enzymes, DNA gel retardation assay and PCR were used to identify apoptin gene stability and to decide the best N/P ratio as well as the model effect in the progress of replication. Human melanoma cells A375 are transiently transformed by nanoparticles containing apoptin gene and apoptosis was measured by MTT assay at various time period. RESULTS: Morphology studies revealed that the particles were spherical in shape with smooth surface. The mean particle diameter ranged from 200-300 nm. The ratio of the chitosan to apoptin DNA (N/P ratio) was 5.5:1. The apoptin gene in chitosan/apoptin nanoparticles could be protected from DNase degradation and could be used as the model in the process of replication. The nanoparticles with apoptin gene could induce apoptosis of A375 cells in dose-dependent manner in vitro at 48 h after transformation. CONCLUSION: The chitosan vector and apoptin gene could be combined to be a safe gene delivery nanoparticles system, which could induce apoptosis in human melanoma cells A375.

[Serum IL-1 receptor antagonist and IL-1beta in patients with Graves' opthalmopathy].

OBJECTIVE: To investigate the mechanism of Graves' ophthalmopathy (GO) and the relationship between serum IL-1 ra and IL-1beta and the effectiveness of glucocoids in treating GO. METHODS: Serum IL-1ra and IL-1beta were measured by Enzyme linked immunosorbent assay (ELISA) and Radioimmunoassay (RIA) in three groups of people, which included 20 patients with severe GO (NOSPACES> or = IV), 20 patients with Graves' disease (GD) without ophthalmopathy, and 20 healthy volunteers. Repeated measurements were undertaken in GO patients before and after glucocoids treatment to analyze the relationship between serum IL-1ra, and IL-1beta and the effectiveness of glucocoids therapy. RESULTS: Before glucocoids therapy,GO patients had higher serum IL-1beta and lower IL-1ra/IL-1beta ratio. Glucocoids therapy increased serum IL-1ra and decreased IL-1beta, and thus increased the IL-1ra/IL-1beta ratio. CONCLUSION: Patients with GO have deficiency in IL-1ra synthesis. Glucocoids therapy can stimulate IL-1ra synthesis and inhibit IL-1beta synthesis.

[Apoptin-induced apoptosis of human melanoma cells A375 via activation of caspase-3].

AIM: To study whether apoptin gene induces apoptosis in human melanoma cells A375 via the activation of caspase-3. METHODS: The human melanoma cells A375 were transiently transfected with recombinant pcDNAA3 plasmid, which contained apoptin gene, and the apoptosis was measured by DNA agarose electrophoresis, RT-PCR and flow cytometry. Caspase-3 relative activity was analyzed by colorimetric assay at various time. RESULTS: At 48 h after transfection, the apoptin gene could induce apoptosis of human melanoma cells A375 in vitro and the mRNA of apoptin gene could be detected by RT-PCR. Caspase-3 activity began to rise at 24 h after transfection and reached the peak at 72 h. CONCLUSION: Apoptin gene can induce apoptosis in human melanoma cells A375 via active caspase-3 and the activity of caspase-3 is time-dependent.