Objective　To compare the changes in the genetic diversity of Anopheles minimus through the research on the population genetic characteristics of Anopheles minimus between different years in Nabang Town, Yingjiang County, Yunnan Province. Methods Anopheles mosquitoes were collected by light traps in Nabang Town, Yingjiang County, Yunnan Province in May 2014 and May 2021. After morphological identification, each mosquito was individually stored in separate tubes for further analysis. DNA of Anopheles minimus was extracted using kits. Microsatellite sequences in the template DNA were amplified using eight pairs of fluorescent primers, and the resulting products were subjected to capillary electrophoresis by a sequencing company. PopGen32 software was used to calculate the observed number of alleles (Na), effective number of alleles (Ne), observed heterozygosity (Ho), expected heterozygosity (He), and Shannon's information index (I) for individual microsatellite loci and population groups. PIC-CALC software was used to calculate the polymorphic information content (PIC). Results A total of 158 mosquitoes belonging to 6 Anopheles species were captured in 2014, while 529 mosquitoes belonging to 5 Anopheles species were captured in 2021. The composition ratio of Anopheles minimus among the mosquito species differed significantly between 2014 and 2021 (χ2=70.48, P<0.01). For 8 microsatellite loci, a total of 85 alleles were detected, a range of 6-20 alleles per locus and an average of 10.625 alleles. Ne ranged from 1.717 to 7.797, with an average of 4.011. The highest PIC was found in the am4 locus, and the lowest in the am25 locus. For the population groups, 77 alleles were found in 2014, and 62 alleles were found in 2021. Ne ranged from 1.630 to 8.658, with an average of 4.147 in 2014. Ne ranged from 1.760 to 6.744, with an average of 3.698 in 2021. The average Ho was 0.641 in 2014 and 0.650 in 2021, while the average He was 0.699 in 2014 and 0.691 in 2021. The Shannon's index ranged from 0.774 to 2.493 in 2014, with an average of 1.579, and from 0.938 to 2.224 in 2021, with an average of 1.464. Na, Ne, I, and PIC were all higher in 2014 compared to 2021, with Na: 9.625>7.750, Ne: 4.147>3.698, I: 1.579>1.464, and PIC: 0.655>0.640, respectively. Conclusions The populations of Anopheles minimus in Nabang Town, Yingjiang County, exhibited high levels of polymorphism in both 2014 and 2021. However, the genetic diversity of the population in 2021 was lower than that in 2014.

Objective To explore the prognosis related genes of pancreatic ductal adenocarcinoma (PDAC)and investigate the molecular regulation mechanism.Methods Gene expression data of 102 PDAC patients with complete clinical survival data were selected from gene expression database of National Center for Biotechnology Information.The 106 transcription regulation gene collection was collected from Transfac database.The 715 microRNA (miRNA)target regulation gene collection was selected according to PicTar and TargetScanS method.Biological pathway data obtained from the Kyoto Encyclopedia of Genes and Genomes (KEGG).The known cancer genes were collected from the cancer gene census (CGC) database.Univariate Cox proportional hazards model was used to analyze the correlation between gene expression data and survival time,then obtained survival related candidate genes from the whole genome. Then the enriched genes were analyzed by hypergeometric distribution algorithm from three databases. Multiple correction testing was performed by BH-FDR method (FDR < 0.05 ).Kaplan-Meier was performed for survival curve analysis of PDAC.Results The results of data of 102 PDAC patients analyzed by univariate Cox proportional hazards model indicated that 273 genes were significantly related to the survival time of patients (P <0.000 1 ).After 273 survival genes were enrichment analyzed in 106 transcription factor regulation gene collection,12 survival genes enriched transcription factor target gene sets were found.After 273 survival genes were enrichment analyzed in 715 miRNA target regulation gene collection,11 survival genes enriched miRNAs target sets were discovered.After 273 survival genes were enrichment analyzed in pathway data of KEGG,15 survival genes enriched pathways were obtained. Period circadian clock 2 (PER2 )was regulated by CCAAT/enhancer binding protein (CEBPA)at transcription level and regulated by miRNA-32 after transcription.The prognosis of PDAC was affected by circadian rhythm pathway.The 102 patients with PDAC were ranked according to the expression of PER2 from high to low,the first 51 cases were included in PER2 higher expression group and the left were included in PER2 lower expression group.Kaplan-Meier survival analysis indicated that PER2 was significantly correlated with prognosis of PDAC (P <0.01 ).Conclusion CEBPA/miRNA-32/PER2 and its related circadian clock pathway may be the target pathway in interfering the development of PDAC,and is correlated with the prognosis of PDAC.

Background Dyslipidemia is one of the major causes of age-related macular degeneration (AMD).At present,the study of the preventive and treating methods of A MD is still a hot spot.Objective The purpose of this study was to observe the effect of tonifying the spleen and promoting blood circulation on the retina and Bruch membrane in apolipoprotein E-deficient (ApoE-/-) mice with dyslipidemia.Methods Thirty-six ApoE-/-mice aged 2 months were randomly divided into the normal diet group,high fat diet group and medicine group.A diet with a higher content of fat was given for 5 consecutive months to the mice of the high fat diet group and medicine group,and in the last month,a concoction that tonifies the spleen and promotes blood circulation was gavagely administered in the medicine group,and an equivalent volumes of normal saline solution was administered in the same way in the normal diet group and high fat diet group.Total plasma cholesterol (TC),low density lipoprotein (LDL)and triglyceride (TG) were detected by (ELISA? Name of assay?) using the 7170 Hitachi automatic biochemical analyzer,and morphological changes of the retina and Bruch membrane were examined by light microscopy and transmission electron microscopy.The number of outer nuclear layer (ONL) cells,retinal pigment epithelial (RPE) cells and the thickness of the Bruch membrane were examined by semi-quantitative histopathology with the Mias 2000 Imaging Analyzer System.Results The concentrations of TC,LDL and TG were (6.47 ±0.49) mmol/L,(1.46 ±0.10)mmol/L and (0.62 ±0.21) mmol/L,respectively,in 7-month-old mice of the medicine group,showing a significant reduction in comparison with (10.53 ±0.30) mmol/L,(1.90±0.13) mmol/L,(1.15±0.29) mmol/L of the high fat diet group,and (9.63 ± 0.18) mmol/L,(1.12 ± 0.15) mmol/L,(0.88 ± 0.21) mmol/L in the normal diet group (P＜0.05-0.01).The disorder and atrophy of ONL and RPE cells,divergence of fiber of the Bruch membranes were found in both the high fat diet group and normal diet control group under the light microscope,and drusen formed in some of the mice in the high fat diet group.However,ONL and RPE were well organized in the medicine group.The cell numbers in the ONL and RPE layer in the 7-month-old mice were (23 124.00±755.18) and (10.75±0.59),respectively,in the medicine group,(19 107.00 ± 1436.82) and (8.55 ± 1.11),respectively,in the high fat diet group,(21 663.00± 1073.27) and (9.75 ±0.58),respectively,in the normal diet group,with significant differences among them (P＜0.05-0.001).Thickness of the Bruch membrane in the medicine group extensively reduced in high fat diet group and normal diet control group (P＜0.01).The ultrastructures of the RPE and Bruch membrane were much more improved in the mdedicine group.Conclusions Tonifying the spleen and promoting blood circulation can attenuate hyperlipemia in ApoE-/-mouse;furthermore,it lessens the pathological abnormalities in the ONL,RPE and Bruch membrane.

ObjectiveTo discuss the experimental method and the mechanisms on treating diseases by acupuncturing the ST36(Zusanli).MethodsUsing Positron Emission Tomography(PET) and functional Magnetic Resonance Imaging(fMRI) to obtain the experimental data about glycometabolism and cerebral blood stream,using SPM and ROI image-analytical method to obtain the visual experimental evidence when acupuncturing the ST36. ResultsThere are certain increases of glycometabolism and cerebral blood stream in ipsilateral hypothalamus and bilateral temporal lobe, when acupuncturing the ST36. Conclusions Acupuncturing the ST36 can lead to the functional changes in vegetative nerve center and temporal lobe, which is close correlated with the therapeutical effects of ST36.

Objective To evaluate the effectiveness of blood gas analysis in samples from two regions.Methods 112 patients with APACHE Ⅱ scores 27 ～ 43 were taken arterial blood from dorsalis pedis artery and femoral artery respectively. Compare the clinical effect of the samples. Results There is no statistically significant difference between results of two blood collection methods ( P ＞ 0. 05 ), While there is statistically significant difference in puncture success ( 110 vs 100, χ2 = 14.23, P ＜ 0. 05 ), vein mistaken injury, hematoma formation, thrombosis, and compression time ( 12 vs 1,7 vs 0,6 vs 0,5.5 ±0.7 vs 2.5 ±0.5 ;χ2 =9.32,5.99,4.85,t = 38. 06, P ＜ 0. 05 ). Conclusions Blood collection method should be determined by the different situation of critically ill patients. Femoral artery blood sample should be taken with caution in order to prevent complication and improve quality of nursing care.

Previous methods used for nuclear transplantation were further investigated to develop a method that was both easy to carryout and did not require any special apparatus, such as Piezoimpact or Spindle-View. Following the puncture of zona pellucida with two holes by injection pipette that contained donor nuclei or cells, the injection pipette was pulled back to the perivitelline space while the negative pressure was increased in the holding pipette until the polar body and karyoplasm were wiped off completely. Then a reconstructed embryo was completed by the direct injection of the donor nucleus or cell without pulling out the injection pipette. 200 oocytes were manipulated using this method and it cost about 40 seconds with nucleus injection and about 30 seconds with cell injection to complete a reconstructed embryo. The success rates were 62.6% and 86. 0%, respectively, and enucleation rate was about 73.3% validated by Hoechst 33342. Using this method, the nucleus was completely eliminated and another was injected using the microscope and micromanipulator. Moreover, the efficiency of nuclear transplantation and survival rate of reconstructed embryos were greatly improved. Furthermore, it is very easy to manipulate and popularize in practice.
Animals ; Cell Culture Techniques ; methods ; Cell Nucleus ; metabolism ; Cells, Cultured ; Cloning, Organism ; methods ; Embryo, Mammalian ; cytology ; metabolism ; Embryonic Development ; Female ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Inbred DBA ; Mice, Inbred Strains ; Nuclear Transfer Techniques ; Oocytes ; cytology ; metabolism ; Zona Pellucida ; metabolism

Background There is a growing interest in studying the relationship between intrinsic resistance and biofilms resistance to drugs. However, the relationship still remains unclear in the macroscopic bacterial growth. Our study is to illuminate the change of bacterial drug resistance of gyrA mutant and active efflux pump during the development of Pseudomonas aeruginosa (P. aeruginosa) biofilms. Methods The strains of type Ⅱ topoisomerase gene mutant (gyrA mutant) and multidrug resistance (MDR) efflux pump were clinical isolates and detected by polymerase chain reaction (PCR). The process of bacterial biofilms development was observed by scanning electron microscope. Triparental mating experiments were performed to transfer report gene of green fluorescent protein (GFP) into P. aeruginosa biofilms strains and followed by analysis of bacterial survival rate between intrinsic resistance and biofilms resistance.Results The fluorescent strains with pGFPuv could develop mature biofilms on Teflon surface. Before a period of 72 hours, the survival rate of biofilms bacteria and intrinsic resistance strains in ciprofloxacin solution was significantly different (P<0.05). The survival number of intrinsic resistance strains (gyrA mutation and active efflux pump) was illustriously higher than biofilm strain in the initial stage of biofilms development. After 72 hours incubation, there was no clearly difference between mutants and biofilms strains in the survival rate (P>0.05). The carbonyl cyanide m-chlorophenylhydrazone and azithromycin could significantly reduce the drug resistance of biofilm strains and efflux pump strains.Conclusions In the development of P. aeruginosa biofilms, the strains of gyrA mutation and MDR efflux could be conferred with new level of drug resistance. When co-cultured mutated strains with biofilm strains, biofilms may play a major role in bacterial resistance. But after 72 hours incubation (a mature biofilms had been developed), there was no clearly difference between the number of mutant strains and biofilm strains.

[Objective]To investigate the etiology,clinical manifestation,treatment and prognosis of optic disc vas-culitis.[Method]Twenty-four eyes of 21 patients were enrolled in this retrospective study.Eye examinations,treatment, and effect were recorded.[Result]Six were male and 15 were female.The age was between 19 and 43 years old(average:28.7±1.6).85.7% of the patients referred to the clinic with mild to moderate decreased vision.Edema of the optic disc can be seen in both types while tortuous veins can also be found in type 2.Similar characteristics were noticed in OCT,FFA, and etc.With a follow-up of 4.52±0.98 months after treatment(prednisone:initial dose 1.0-1.2 mg/kg),the BCVA of the affected eyes improved significantly.[Conclusion]Optic disc vasculitis is affected by autoimmune disorder,infection,hy-perlipidemia,and etc.Edema of the optic disc with/without tortuous veins and retinal hemorrhage can be noticed.Similar diseases should be excluded in avoidance of misdiagnosis. Systemic examination and complete solution should be per-formed.Glucocorticoid helps to improve the visual function.The application of anti-VEGF is effective in secondary macu-lar edema.However,the long-term efficacy is awaiting being confirmed.

Paeoniflorin (PF) is the chief active component of paeonia, with diverse pharmacological actions and wide application. Recently, the effect of PF on nervous system has attracted increasingly more attention. According to current study findings, PF can ameliorate the decline of memory and learning capacities in many dementia model animals, and have effect in protecting the cerebral ischemia injury, treating Parkinson's disease, reliving pain and improving neural synapse plasticity. Thought its mechanism has not been clarified, current findings show that adenosine A1 receptor plays an important role, while M cholinergic receptor, opiate receptor, calcium ion channel and NF-KB may also play a part in paeoniflorin's effect on nervous system.
Animals ; Benzoates ; pharmacology ; Bridged-Ring Compounds ; pharmacology ; Calcium Channels ; metabolism ; Glucosides ; pharmacology ; Learning ; drug effects ; Memory ; drug effects ; Monoterpenes ; NF-kappa B ; metabolism ; Nervous System ; drug effects ; metabolism ; Receptor, Adenosine A1 ; metabolism

Objective: To investigate the value of initial CT quantitative analysis of ground-glass opacity (GGO), consolidation, and total lesion volume and its relationship with clinical features for assessing the severity of coronavirus disease 2019 (COVID-19). Materials and Methods: A total of 84 patients with COVID-19 were retrospectively reviewed from January 23, 2020 to February 19, 2020. Patients were divided into two groups: severe group (n = 23) and non-severe group (n = 61). Clinical symptoms, laboratory data, and CT findings on admission were analyzed. CT quantitative parameters, including GGO, consolidation, total lesion score, percentage GGO, and percentage consolidation (both relative to total lesion volume) were calculated. Relationships between the CT findings and laboratory data were estimated. Finally, a discrimination model was established to assess the severity of COVID-19. Results: Patients in the severe group had higher baseline neutrophil percentage, increased high-sensitivity C-reactive protein (hs-CRP) and procalcitonin levels, and lower baseline lymphocyte count and lymphocyte percentage (p < 0.001). The severe group also had higher GGO score (p < 0.001), consolidation score (p < 0.001), total lesion score (p < 0.001), and percentage consolidation (p = 0.002), but had a lower percentage GGO (p = 0.008). These CT quantitative parameters were significantly correlated with laboratory inflammatory marker levels, including neutrophil percentage, lymphocyte count, lymphocyte percentage, hs-CRP level, and procalcitonin level (p < 0.05). The total lesion score demonstrated the best performance when the data cut-off was 8.2%. Furthermore, the area under the curve, sensitivity, and specificity were 93.8% (confidence interval [CI]: 86.8–100%), 91.3% (CI: 69.6–100%), and 91.8% (CI: 23.0–98.4%), respectively. Conclusion CT quantitative parameters showed strong correlations with laboratory inflammatory markers, suggesting that CT quantitative analysis might be an effective and important method for assessing the severity of COVID-19, and may provide additional guidance for planning clinical treatment strategies.