ObjectiveTo explore the potential mechanism underlying electroacupuncture therapy with the midnight-noon ebb-low method with hour-prescription for chronic obstructive pulmonary disease （COPD） with lung-qi deficiency syndrome from the perspective of brain and muscle basic helix-loop-helix ARNT-like 1 （BMAL1） as well as neutrophil extracellular traps （NETs）. MethodsThe experiment was conducted in two phases. For the therapeutic effectiveness observation， 24 rats were randomly allocated into control group 1， model group 1， midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group， with 6 rats per group. Rat models of COPD with lung-qi deficiency syndrome were established via cigarette smoke exposure combined with intratracheal instillation of lipopolysaccharide （LPS）. After successful modelling， rats in midnight-noon electro-acupuncture group 1 received electro-acupuncture at acupoints "Taiyuan （LU 9）"， "Feishu （BL 13）" and "Zusanli （ST 36）" during 05：00—07：00， 30 minutes per treatment once every other day for 14 consecutive days （7 sessions in total）. Rats in the conventional electro-acupuncture group received identical electroacupuncture manipulation at random daytime hours （08：00—18：00）. Pulmonary function parameters including forced expiratory volume in 0.3 second （FEV_0.3）， forced vital capacity （FVC） and FEV_0.3/FVC ratio were detected. Pulmonary histopathological changes were observed via hematoxylin-eosin （HE） staining. Plasma levels of pro-inflammatory factors interleukin-1β （IL-1β） and tumor necrosis factor-α （TNF-α）， as well as pulmonary reactive oxygen species （ROS） content， BMAL1 protein expression and the levels of neutrophil extracellular traps （NETs） biomarkers such as myeloperoxidase （MPO）， neutrophil elastase （NE）， and citrullinated histone H3 （CitH3） in lung tissue were determined. For the mechanistic verification phase， 36 rats were divided into control group 2， model group 2， midnight-noon electro-acupuncture group 2， overexpression empty group， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group， with 6 rats in each group. At the 5th week of model construction， rats in the overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group received intratracheal instillation of 100 μl adenoviral suspension carrying overexpressed Bmal1 gene. Rats in the overexpression empty group were injected with an equal titer and equal volume of blank adenovirus without the Bmal1 coding sequence into the lung at the identical time point. Midnight-noon electro-acupuncture group 2 and overexpression with midnight-noon electro-acupuncture group were consistent with the aforementioned protocol. Finally， pulmonary levels of MPO， NE and CitH3 were measured in all groups. ResultsCompared with the control group 1， the model group 1 exhibited decreased FEV_0.3， FVC and FEV_0.3/FVC ratio， elevated plasma IL-1β， TNF-α and pulmonary ROS levels， downregulated lung BMAL1 protein expression， and increased contents of MPO， NE and CitH3 （all P＜0.05）； typical NETs-like structures with sparse granular substances attached to fibrous networks were observed under scanning electron microscopy； HE staining revealed damaged alveolar architecture accompanied by inflammatory cell infiltration. Compared with model group 1， both the midnight-noon electro-acupuncture group 1 and conventional electro-acupuncture group achieved obvious improvements in all above indicators， with superior therapeutic effects in midnight-noon electro-acupuncture group 1 （P＜0.05）. Consistently， midnight-noon electro-acupuncture group 1 showed more prominent alleviation of NETs-like structure formation and lung pathological injury compared to the conventional electro-acupuncture group. Mechanistic validation results demonstrated that compared with model group 2， the average optical density of MPO as well as the protein levels of MPO， NE and CitH3 were markedly reduced in midnight-noon electro-acupuncture group 2， overexpression BMAL1 group and overexpression with midnight-noon electro-acupuncture group （P＜0.05）； furthermore， these parameters were significantly lower in the overexpression with midnight-noon electro-acupuncture group than in midnight-noon electro-acupuncture group 2 （P＜0.05）. ConclusionElectro-acupuncture based on the midnight-noon ebb-low method with hour-prescription may alleviate pulmonary inflammation in COPD by upregulating the expression of clock protein BMAL1 and inhibiting excessive accumulation of NETs in lung tissue.

Objective:To analyze the clinical characteristics of monkeypox patients.Methods:The clinical data and laboratory findings of 4 patients with monkeypox patients diagnosed at Yiwu Central Hospital in July 2023 were analyzed. Herpes fluid and skin tissue samples were collected, the viruses were isolation and cultured in African green monkey kidney cells (Vero) and identified with whole gene sequencing.Results:All four patients were male, aged 24-35 years. All patients had male-to-male behavior within 21 days before onset of the disease. Among them, one patient has AIDS and one patient has syphilis. Four patients presented with perineal skin lesions with itching, and 3 patients were found to have enlarged lymph nodes upon admission. Laboratory testing: lymphocyte abnormality (4.57×10 9/L) in 1 case; increased procalcitonin (0.25 ng/mL) in 1 case; elevated IL-10 levels ( 7.11 ng/L and 9.42 ng/L) in 2 cases; increased IL-6 (66 ng/L) and IL-4 (3.24 ng/L) in 1 case, respectively. One case had abnormal myocardial zymogram with a elevated lactate dehydrogenase level of 313 U/L. The monkeypox virus was isolated from lesion tissue and herpes fluid, and the whole gene sequencing identified it as the B. 1.3 subtype of the IIb evolutionary branch, exhibiting typical pathological effects on Vero cells. Conclusion:The clinical manifestations of the 4 monkeypox patients confirmed in Zhejiang province are mild, patients had a definitive history of male-to-male sexual behavior and the virus strains belong to the B. 1.3 lineage of the IIb evolutionary branch.

A high throughput measurement method of human red blood cells (RBCs) deformability combined with optical tweezers technology and the microfluidic chip was proposed to accurately characterize the deformability of RBCs statistically. Firstly, the effective stretching deformation of RBCs was realized by the interaction of photo-trapping force and fluid viscous resistance. Secondly, the characteristic parameters before and after the deformation of the single cell were extracted through the image processing method to obtain the deformation index of area and circumference. Finally, statistical analysis was performed, and the average deformation index parameters (
Erythrocyte Deformability ; Erythrocytes ; Humans ; Microfluidics ; Optical Tweezers ; Viscosity

The contractile force of hepatic stellate cells plays a very important role in liver damage, hepatitis and fibrosis. In this paper, a method based on polydimethylsiloxane (PDMS) thin micropillar arrays is proposed to measure the contractile force of human hepatic stellate cell line LX-2, which enables dynamic measurement of the subcellular distribution of force magnitude and direction. First, thin micropillar arrays on glass bottom dish were fabricated using two-step casting process in order to meet the working distance requirement of 100× objective lens. After hydrophilic treatment and protein imprint, cells were seeded on the micropillar arrays. LX-2 cells, which were quiesced by growth in serum-free medium, were activated by adding fetal bovine serum (FBS). The deflections of the micropillars were achieved by image processing technique, and then the contractile force of cells exerted on the micropillars was calculated according to mechanical simulation results, and was analyzed under both quiescent and activated conditions. The experimental results show that the average traction force of quiescent cells is about 20 nN, while the contractile force of activated cells increased to 110 nN upon adding FBS. This method can quantify the contractile force of LX-2 cell on subcellular scale in both quiescent and activated states, which may benefit pathology study and drug screen for chronic liver diseases resulted from liver fibrosis.
Cell Line ; Hepatic Stellate Cells ; cytology ; Humans ; Image Processing, Computer-Assisted ; Mechanical Phenomena

The traditional method of multi-parameter flow data clustering in flow cytometry is to mainly use professional software to manually set the door and circle out the target cells for analysis. The analysis process is complex and professional. Based on this, a clustering algorithm, which is based on -distributed stochastic neighbor embedding ( -SNE) algorithm for multi-parameter stream data, is proposed in the paper. In this algorithm, the Euclidean distance of sample data in high dimensional space is transformed into conditional probability to represent similarity, and the data is reduced to low dimensional space. In this paper, the stained human peripheral blood cells were treated by flow cytometry, and the processed data were derived as experimental sample data. The -SNE algorithm is compared with the kernel principal component analysis (KPCA) dimensionality reduction algorithm, and the main component data obtained by the dimensionality reduction are classified using -means algorithm. The results show that the -SNE algorithm has a good clustering effect on the cell population with asymmetric and trailing distribution, and the clustering accuracy can reach 92.55%, which may be helpful for automatic analysis of multi-color multi-parameter flow data.

Objective To evaluate the performance of Zika virus (ZIKV) disease prevention and control.Methods Descriptive epidemiological analysis was conducted on the clinical manifestations,laboratory detection results and disease progression of the third imported ZIKV disease case in the mainland of China.Results On 19 February 2016,a ZIKV disease case was confirmed in Yiwu,Zhejiang province,which was the third imported case of ZIKV disease confirmed by China CDC laboratory and expert consulting.The patient just had a travel to Fiji and Samoa and had mosquito bite history in Samoa.The patient was hospitalized on 16 February after the onset on 14 February and the eruption on 15 February.The body temperature of the patient became normal on 17 February,the rash disappeared on 19 February and the conjunctivitis resolved on 20 February.The positive detection of the viral nucleic acid in blood was only for 3 consecutive days,and the viral nucleic acid could be detected in urine even after negative detection in blood for 4 days.Conclusion The symptoms of the patient were typical.ZIKV can be detected by using blood sample in early phase,but after body temperature become normal,the virus can be detected in urine.

Objective To assess the correlation between inflammation,specific immune response and coronary heart disease(CHD). Methods Thirty healthy cases passed the health examination were taken as the control group. Eighty cases who were diagnosised into CHD,affirmed by coronary angiography,were divided into three groups: acute myocardial infarction (AMI) group(26 cases),unstable angina pectoris (UP) group (24 cases),and stable angina pectoris (SP) group(30 cases). All the cases were tested on the concentrations of C-reactive protein(CRP),IgA,IgG,IgM in serum. Results The serum indices of CRP,IgG,IgA in AMI group and UP group were significantly difference than those in the control group (P0.05). Conclusion The correlation between inflammation and immune system activation are closely associated with CHD.

BACKGROUNDTo identify the relationship of cervical cancer with pathogen infectious, cytokine and Se.

METHODSOn the one hand regarded tissues of the carcinoma of the uterine cervix with 195 cases as the experimental group and ordinary cervical tissues with 75 cases as the control group. Polymerase chain reaction (PCR) to detect them. On the other hand applied ELISA to detect cytokine IL-2R, TNF and fluorescent luminosity technique to detect element Se in the serums.

RESULTSIn the experimental group those infected pathogen were 166 cases (85.1%), In all of pathogen HPV 16,18,35 type were 60 cases (30.8%), HSV 2 were 60 cases (30.8%), While those ordinary tissues infected pathogen were 15 cases (20 0%),in the contrast group. HPV 16,18,35 type and HSV 2 mere 4.0% and 6.7% respectively,(P<0.001). In the serums of effective 62 experimental objects IL-2R (x=356.44 U/ml) and TNF (x=373.48 pg/ml) were much high than them in the serums of effective 36 contrast objects (P<0.001). But Se (x=0.058 mg/ml) was lower than it in the serums of contrast objects (P<0.05).

CONCLUSIONSThe occurrence of carcinoma of the uterine cervix is closely connected with infection of HPV 16,18,35 and HSV2, high level of cellular factors IL-2R, TNF and low level of element Se.

Female ; Herpes Simplex ; virology ; Humans ; Papillomavirus Infections ; virology ; Polymerase Chain Reaction ; Receptors, Interleukin-2 ; blood ; Selenium ; blood ; Tumor Necrosis Factor-alpha ; metabolism ; Tumor Virus Infections ; virology ; Uterine Cervical Neoplasms ; blood ; virology

OBJECTIVETo evaluate the etiology of Shiga like toxin producing Escherichia coli (SLTEC) in children with diarrhea.

METHODSWe designed and synthesized 3 pairs of primers located in the SLT1, SLT2, and eaeA genes of enterohaemorrhagic Escherichia coli (EHEC), while the virulent genes SLT1, SLT2, and eaeA from E.coli species were amplified by polymerase chain reaction (PCR).

RESULTSOne strain of EHEC with SLT1, SLT2, and eaeA in 29 reference strains of diarrhea-causing E.coli (DCEC) and 10 strains of other enterobacteria detected by PCR had positive reactions, while all other DCEC and enterobacteria were negative. Of 474 strains of E. coli isolated from 1032 children with diarrhea and detected by PCR, 20 strains of SLT1 producing E. coli (4.2%) positive, and 7 strains of SLT2 producing E. coli (1.5%) positive; while of 74 strains of entero-SLTs-producing and invasive Escherichia coli (ESIEC), 15 strains of SLT1 (20.3%) and 5 strains of SLT2 (6.8%) were positive.

CONCLUSIONShiga-like toxin E. coli has been identified as a major etiologic agent of children with diarrhea in Taiyuan, China.

Child ; Diarrhea ; microbiology ; Escherichia coli ; classification ; isolation & purification ; pathogenicity ; Feces ; microbiology ; Humans ; Polymerase Chain Reaction ; methods ; Sensitivity and Specificity ; Shiga Toxins ; genetics