By organizing literature, summarizing and analyzing the relevant experimental and clinical studies, this article analyzed the study status of the analgesic action of electroacupuncture with different frequencies from the chemical mechanism, neuro- physical mechanism, molecular mechanism, and action mechanism. It brought up that electroacupuncture of different frequencies can activate different regions in the central system and generate different central opioid peptides, excite different receptors and afferent neurons, and different acupuncture manipulations can induce different neural electrical information encoding sequences. This article has provided theoretical evidences for future researches on the mechanism of acupuncture analgesia.

BACKGROUND: Inhibiting the degradation of extracellular matrix in the intervertebral disc can delay the degenerative process of intervertebral disc. Matrix metalloproteinases-3 (MMP3) is considered as a key enzyme for degradation of extracelular matrix components such as type II collagen and aggrecan.
OBJECTIVE:To construct the short hairpin RNA lentiviral vector targeting human MMP3 gene and to detect its efficiency of gene silence by infecting human degenerative nucleus pulposus cells.
METHODS:According to the human MMP3 mRNA (NM_002422.4) sequence, four groups of the short hairpin RNA gene sequences targeting MMP3 were designed, synthesized and annealed to form double stranded DNA fragments, which were connected with the LV3 vectors digested by BamHI andEcoRI enzymes, and then transfected into the competent cels. The positive clones were identified by PCR, and analyzed by sequencing. The packaging and titer of lentivirus were determined after transfecting 293T cells. Human degenerative nucleus pulposus cels were infected with lentivirus vector, and the transfection efficiency of each group was observed under inverted fluorescence microscope. The interfering efficiency was detected by real time-PCR and western blot at 72 and 96 hours.
RESULTS AND CONCLUSION:The ds-oligo DNA was successfully inserted into the lentiviral vector as confirmed by electrophoresis and sequence analysis. The recombinant lentivirus was harvested from 293T cels with a viral titer of 1-5 ×108 TU/mL. RNA interference targeting the GCC AGG CTT TCC CAA GCA AAT sequences with the highest interfering efficiency in MMP3 gene at 72 and 96 hours resulted in suppression of MMP3 mRNA expression by 98% and 72%, respectively; and at 96 hours, the interfering efficiency of protein expression was 57.2%. The recombinant lentivirus vector containing RNA interference targeting MMP3 gene is successfuly constructed, which lays a foundation for further studies on the MMP3 function and gene therapy.

Objective:To systematically evaluate the volumetric changes of upper airway after rapid maxillary expansion using Meta analysis.Methods:All literatures about volumetric changes in the upper airway after rapid maxillary expansion were searched from the database in general.The literatures were screened according to the correlation and the inclusion criteria,included the literatures were analyzed by Rev Man 5.3.Results:11 literatures were finally selected.Meta analysis indicated that,after treatment nasopharyngeal volume increased by 0.62 cm3(P=0.000 2),palatopharyngeal volume increased by 0.62 cm3(P=0.02),glossopharyngeal volume increased by 0.39 cm3(P=0.11),oropharyneal volume increased by 0.40 cm3(P=0.27).Conclusion:The existing evidence indicates that rapid maxillary expansion can increase the volume of nasopharynx and retropalatal part of upper airway.

Abstract: Objective　To analyze the results of the surveillance of hand, foot and mouth disease (HFMD) pathogens in Baoshan City, Yunnan Province in 2022, and to analyze the genetic characteristics of the main epidemic strain coxsackievirus A16 (CVA16), in order to provide scientific basis for the prevention and control of HFMD. Methods　Samples of hand, foot and mouth disease cases in Baoshan City submitted to Yunnan Province in 2022 were selected, the samples were processed, and the viral nucleic acid was extracted, and the Enterovirus (EV) VP4/VP2 binding region gene was amplified with primers (MD91/OL68-1) and sequenced. The sequence was BLAST searched in GenBank to determine the virus type, and then the full sequence of VP1 region was amplified with relevant primers of each type of virus and sequenced. Viruses were identified by using Enterovirus Genotyping Tool Version 1.0. according to the reference documents, the reference sequences were downloaded for making the phylogenetic tree, and the test results were statistically analyzed. Results　A total of 307 clinical samples of hand-foot-mouth disease in Baoshan City in 2022 were detected by real-time RT-PCR. There were 280 laboratory-confirmed cases (91.21%), among which the detection rate of CVA16 was 55.05% (169/307), EV-A71 was 3.58% (11/307), and other enteroviruses were 32.57% (100/307). Of the 206 HFMD test specimens, the VP4/VP2 junction and VP1 gene sequences of enterovirus were amplified and identified, and 29 enterovirus strains were obtained, with a positive virus rate of 14.08% (29/206). All viruses belonged to group A and were divided into 3 serotypes, which included 27 strains of CVA16 (93.10%, 27/29), 1 strain of EV-A71 (3.45%, 1/29), and 1 strain of CVA10 (3.45%, 1/29). Group B, C, and D viruses were not detected. The positive rate of VP4/VP2 binding region and VP1 region of coxsackievirus A16 gene in Baoshan City in 2022 was 13.11% (27/206). The genetic evolution analysis showed that the 27 strains of CVA16 belong to B1a subtype and can be divided into two evolutionary branches. Conclusions　In 2022, the main epidemic strain of HFMD in Baoshan City, Yunnan Province is CVA16 gene, belonging to B1a subtype, which can be divided into two branches, indicating 2 transmission chains prevalent in Baoshan City. Future efforts should focus on strengthening surveillance, improving the quality of monitoring, and understanding the characteristics of viral prevalence..

OBJECTIVETo screen long non-coding RNA (lncRNA) associated with radiosensitivity in colorectal carcinoma cell lines.

METHODSColony formation assay was performed in colorectal cancer cell lines HT29, SW480, RKO, Lovo and HCT116 after irradiation with different radiation doses. Radiation sensitivity of these 5 cell lines was detected through survival fraction at 2 Gy (SF2 value). High-throughput lncRNA chip was used to screen lncRNA genes with expression differences more than 2 folds among SW480, RKO and Lovo. Further experiment on the expression differences of lncRNAs selected was conducted by realtime PCR.

RESULTSThe radiosensitivity order of these 5 cell lines from low to high (SF2 value from high to low) was HT29 (0.83 ± 0.03), SW480 (0.69 ± 0.02), RKO(0.53 ± 0.02), Lovo (0.47 ± 0.05), HCT116 (0.32 ± 0.03) (P < 0.01). Five lncRNAs associated with radiation sensitivity were screened. Among them, expression levels of R05532, NR_015441, and NR_033374 were positively correlated with radiation resistance(all P < 0.01), and expression levels of the other 2 lncRNAs, NR_073156 and AA745020, were not correlated with radiation resistance of colorectal cancer cells (both P>0.05).

CONCLUSIONSlncRNA R05532, NR_015441 and NR_033374 may be used as the predictive marker of radiosensitivity of colorectal cancer cells. Higher expression of these genes shows radiation resistance.

Objective:To investigate the influencing factors of urinary tract infection in pregnant women with diabetes and the distribution characteristics of pathogens in the middle urinary tract.Methods:A total of 220 patients with gestational diabetes who visited the Second People′s Hospital of Lianyungang City from December 2018 to December 2021 were selected as the study subjects, and the incidence of urinary tract infection was counted. According to the diagnosis results of urinary tract infection, they were divided into infected group and uninfected group. The infected group took the middle urine for pathogen culture, and the resistance rate of main gram-negative bacteria to antibiotics was analyzed; Logistic regression model was used to analyze the influencing factors of urinary tract infection in pregnant women with diabetes.Results:There were 32 cases of urinary tract infection in 220 patients with gestational diabetes, and the infection rate was 14.55%(32/220). 43 strains of pathogenic bacteria were identified, mainly gram-negative bacilli [72.09%(31/43)], followed by gram-positive cocci [20.93%(9/43)] and fungi [6.98%(3/43)]. Amongthe main gram-negative bacteria, escherichia coli had a high resistance rate to ampicillin and levofloxacin, while Klebsiella pneumoniae had a high resistance rate to ampicillin and cefazolin; There were significant differences between the infected group and the non infected group in age, hospital stay, personal urinary tract infection history, pregnancy sexual life history, use of antibiotics, fasting blood sugar, serum albumin, and glycated hemoglobin (all P<0.05); Multivariate logistic regression results showed that personal history of urinary tract infection, sexual life during pregnancy, non-standard use of antibiotics, serum albumin<30 g/L, glycated hemoglobin ≥7%, and fasting blood sugar ≥8.5 mmol/L were independent risk factors for urinary tract infection in pregnant diabetes patients (all P<0.05). Conclusions:There is a high incidence of urinary tract infection in patients with gestational diabetes, and the risk factors are complex. Gram negative bacilli are the main pathogenic bacteria. Antibacterial drugs can be reasonably selected for intervention according to drug sensitivity test in clinical practice.

OBJECTIVETo study the chemical constituents of the leaves of Psidium guajava.

METHODThe chemical constituents were isolated by column chromatography on silica gel, Sephadex LH-20 and MPLC. Their structures were elucidated on the basis of spectral analysis.

RESULTNine compounds were isolated from this plant, and the structure of them were identified as ursolic acid (1), 2alpha-hydroxyursolic acid (2), 2alpha-hydroxyoleanolic acid (3), morin-3-O-alpha-L-arabopyranoside (4), quercetin (5), hyperin (6), myricetin-3-O-beta-D-glucoside (7), quercetin-3-O-beta-D-glucuronopyranoside (8), 1-O-galloyl-beta-D-glucose (9).

CONCLUSIONCompounds 3, 7-9 were isolated from this plant for the first time.

Objective To analyze the genetic characteristics of VP1 3'region of human coxsack-ievirus B2 (CV-B2) strains isolated from Yunnan province. Methods RT-PCR and gene sequencing were performed to analyze the VP1 3'region of 15 CV-B2 strains isolated from acute flaccid paralysis ( AFP) cases during 2005 to 2006, healthy children in 2013 and hand, foot and mouth disease (HFMD) cases in 2014 in Yunnan province. CV-B2 VP1 gene reference sequences were downloaded from the Genbank. Nucleotide (nt) and amino acid (aa) diversities were calculated by MEGA5. 2 software and a phylogenetic tree was constructed. Genetic and molecular epidemiological characteristics of CV-B2 strains circulating in Yunnan province were analyzed. Results A total of 15 CV-B2 strains were isolated, which were one from 232 AFP cases in 2005, one from 240 AFP cases in 2006, 12 from 400 healthy children in 2013 and one from 500 HFMD cases in 2014. Phylogenetic analysis of the 15 CV-B2 strains in Yunnan province and those down-loaded from the GenBank showed that CV-B2 could be genetically divided into five genotypes. The prototype strain Ohio-1 and one strain (01-1) isolated in Taiwan in 1988 belonged to genotype 1. Strains isolated in France in 2006, 2007 and 2010 belonged to genotype 2. Strains isolated in Yunnan, Shandong, Henan, Fu-jian and Taiwan belonged to genotype 3. Strains isolated in Russia, Yunnan AFP cases in 2005 and 2006 and India belonged to genotype 4. Strains isolated in Taiwan, Shandong and New South Wales, Australia be-longed to genotype 5. Different genotypes distributed in different countries/areas with some confined within specific countries/areas. Conclusions The 12 strains isolated from healthy children and one from HFMD cases in Yunnan province belonged to genotype 3, while the two strains isolated from AFP cases belonged to genotype 4. Diversities in nt and aa sequences between the strains isolated from the healthy children and HFMD case were only 0. 76％ and 0. 03％, respectively, indicating that they might come from the same transmission source. However, the nt and aa diversities between the isolates of genotype 3 ( from healthy children and HFMD case) and genotype 4 (from AFP cases in 2005 and 2006) were 15. 11％-15. 22％ and 2. 76％-2. 72％, respectively. Correlation of CV-B2 with AFP and HFMD was worthy of further study.

Objective:To analyze the etiology of hand, foot and mouth disease (HFMD) cases collected from Wenshan prefecture from 2014 to 2018 and the molecular epidemiology of coxsackievirus A6(CV-A6).Methods:Viruses were isolated by RD cells and Hep-2 cells from stool samples collected from HFMD patients in Wenshan prefecture from 2014 to 2018. Virus RNA was extracted and virus VP4/VP2 junction region sequence was firstly amplified and sequenced by MD91 and OL68-1 primer pairs, then the virus serotype was determined. Virus entire VP1 gene sequences were determined by relative primer pairs according to the references. The reference sequences of CV-A6 virus entire VP1 gene were downloaded from the GenBank and the phylogenetic tree was constructed and the genetic characteristics and molecular epidemiology were analyzed.Results:During five years of study period, a total of 581 strains of enteroviruses (EVs) was isolated with an isolation rate of 20.40% (581/2 848). Among 581 strains, 74 strains were CV-A6, accounting for 12.74% (74/581); 124 were CV-A16, accounting for 21.34% (124/581); 374 were EV-A71, accounting for 64.37% (374/581); nine were other EVs, accounting for 1.55% (9/581). The entire VP1 sequences of 74 CV-A6 strains were filtered by constructing a phylogenetic tree and the completely same strains were excluded from analysis. We finally analyzed the phylogenetic characteristics of 22 strains isolated in this study with 52 reference strains. The results showed that all 22 Wenshan strains belonged to D3a sub-genotype, of which 21 strains belonged to cluster 1, and only one strain belonged to cluster 2.Conclusions:From 2014 to 2018, the outbreaks of HFMD in Wenshan prefecture were mainly caused by EV-A71, CV-A16 and CV-A6, accounting for 64.37%, 21.34% and 12.74% respectively. Phylogenetic analysis showed, similar to the situation in China, the sub-genotype D3a of CV-A6 was the predominant virus and the cluster 1 was the main sub-genotype in this outbreak.

Infectious diseases are still one of the leading causes of morbidity and death globally, affecting public health and life, social and economic development, and even national security. Early detection focuses on detecting the abnormal information of infectious disease outbreaks or epidemics in a timely and sensitive way to conduct field investigation and verification. It is also a precursor to effective surveillance and early warning system. The effective surveillance and early warning system can fully and accurately understand the real conditions, driving forces, and transmission chain of the occurrence of a specific infectious disease outbreak and epidemic and put forward scientific and effective prevention and control strategies and measures. Due to the measurement of the resources support and the particular data collection value, it is not easy to obtain epidemiological, etiological, and other data information in a timely, complete and accurate manner. This paper summarized the theory and technology on early detection, effective surveillance, and early warning information on infectious diseases. It also integrated and utilized the multi-source data, including effective infectious disease surveillance and the country's early warning system, to better understand the outbreak epidemic, causes, risks, processes, and driving forces. Thus, it is possible to set up a sensitive, specific staging measurement innovative technical system to monitor, early warning, and timely respond to acute infectious diseases through multidisciplinary cooperation in China. It provides the basis for strengthening the surveillance and early warning of new emerging and major infectious diseases and public health emergencies, avoiding the spread of inadequate response to infectious disease, and preventing the resources waste of over-response.