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.

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.

[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.

[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.

[Mechanism of apoptin-induced apoptosis of human lymphoma cells U937].

AIM: To study the role of c-Jun N-terminal kinase (JNK) in the apoptosis induced by apoptin gene in human lymphoma cell U937. METHODS: The U937 cells were transiently transfected by pcDNAA3 plasmids containing apoptin gene. Apoptosis of U937 cells was measured by flow-cytometry. Activation of JNK signal pathway was detected by Western blot. RESULTS: Apoptin could induce apoptosis of U937 cells in vitro at 48 h after transfection. The level of phosphorylated JNK was increased at 24 h and reached the peak level at 48 h after transfection. CONCLUSION: Apoptin gene can induce apoptosis of U937 cells in which JNK signal pathway plays an important role.