BACKGROUND:Wogonin is a flavonoid extracted from the root of Scutellaria baicalensis.Previous studies have shown that baicalein has protective effects against cerebral ischemia-reperfusion injury,and can also reduce blood sugar and complications in diabetic mice,but its role and mechanism in diabetic cerebral infarction remain unclear. OBJECTIVE:To explore the effect of wogonin on nerve injury in rats with diabetic cerebral infarction and its mechanism. METHODS:Sprague-Dawley rats were randomly divided into six groups:control group,model group,low-dose wogonin group,medium-dose wogonin group,high-dose wogonin group,and high-dose wogonin+Ras homolog gene family member A(RhoA)activator group.Except for the control group,the other rats were established with diabetes and cerebral ischemia models using intraperitoneal injection of streptozotocin and middle cerebral artery occlusion.Low,medium-and high-dose wogonin groups were intragastrically given 10,20,40 mg/kg wogonin,respectively;high-dose wogonin+RhoA activator group was intragastrically given 40 mg/kg wogonin and intraperitoneally injected 10 mg/kg lysophosphatidic acid;control group and model group were given the same amount of normal saline once a day for 7 consecutive days.Rats in each group were evaluated for neurological deficits and their blood glucose levels were measured after the last dose.TTC staining was applied to detect the volume of cerebral infarction.Hematoxylin-eosin staining was applied to observe pathological changes in brain tissue.ELISA kit was applied to detect tumor necrosis factor-α,interleukin-6,malondialdehyde,and superoxide dismutase levels in brain tissue.Western blot was applied to detect the protein expression of RhoA and Rho-associated protein kinase(ROCK)2 in brain tissue. RESULTS AND CONCLUSION:Compared with the control group,the neuronal structure of rats in the model group was severely damaged,with cell necrosis and degeneration,the neurological deficit score,blood glucose level,and infarct volume were significantly elevated(P<0.05),the levels of tumor necrosis factor-α,interleukin-6,and malondialdehyde,and the protein expression of RhoA and ROCK2 in brain tissue were significantly increased(P<0.05),and the superoxide dismutase level was decreased(P<0.05).Compared with the model group,the low-,medium-,and high-dose wogonin groups showed improved neuronal damage,reduced cell degeneration and necrosis,a significant reduction in neurological deficit score,blood glucose level,infarct volume,and the levels of tumor necrosis factor-α,interleukin-6,and malondialdehyde,and the protein expression of RhoA and ROCK2 in brain tissue,and an increase in the superoxide dismutase level(P<0.05).Compared with the high-dose wogonin group,the high-dose wogonin+RhoA activator group significantly weakened the improvement in the above indexes of rats with diabetic cerebral infarction(P<0.05).To conclude,wogonin can improve the blood glucose level in rats with diabetic cerebral infarction,reduce cerebral infarction and nerve injury,and its mechanism may be related to the inhibition of RhoA/ROCK signaling pathway.

Mental state is an important part of the normal life activities of the human body, and it is also the most external expression and the most easily obtained information of the physical condition. The normal activities of the mind depend on the normal operation of the viscera, qi, and blood, and are a unified whole that prospers together and suffers together. The theory of the five-spirit-viscera in the Yellow Emperor’s Inner Classic revealed that the normal mental activities of the human body were dominated by the five internal organs, that is, the five internal organs were the body and the five spirits were the function. And it highlighted the viewpoint that the five internal organs store the spirits and are actually one. The heart governs the spirit and belongs to the four internal organs. On this basis, Synopsis of Golden Chamber used the internal organs to diagnose and treat mental diseases, integrating the theory of the five spirits into it, forming a unique method of diagnosis and treatment with the heart as the leading factor and regulating the qi and blood of the four internal organs. It identified the pathogenesis of diseases such as pathogenic crying, lily disease, and hysteria from five levels: heart deficiency and weak qi, heart-lung disharmony, heart-liver disharmony, the heart of the loss of the spleen nourishment, and disharmony between heart and kidney. The treatment was mainly to replenish the deficiency of the viscera and eliminate the pathogens, reflecting the characteristics of regulating the mind and calming the four internal organs. This unique view on diagnosis and treatment has profoundly influenced the diagnosis and treatment theories of mental illnesses by later doctors, and is of great significance to the current clinical treatment of such illnesses.

Objective:To construct and test the reliability and validity of an assessment scale for self-management of patients with dry eye, so as to help patients establish dry eye related health behaviors.Methods:Based on the knowledge attitude and practice theory, the initial scale was constructed on the basis of literature review and qualitative interview, and revised through Delphi expert consultation and expert argumentation. After the preliminary investigation, 211 dry eye patients treated in the dry eye nursing outpatient department of Beijing Tongren Hospital from August 2023 to January 2024 were selected by convenience sampling method for questionnaire investigation to test the reliability and validity of the scale.Results:Among 211 dry eye patients, there were 58 males and 153 females, aged (39.31 ± 12.81) years old. The scale of self-management of patients with dry eye included 27 items in 3 dimensions: knowledge, attitude and behavior. The authority coefficient of the two rounds of expert consultation was 0.86, and the Kendall coordination index increased from 0.175 to 0.223 (both P < 0.01). The cumulative variance contribution rates of the three dimensions were 41.843%, 54.219% and 44.273%, respectively. The Cronbach α coefficient of the scale was 0.787, and the Spearman-Brown coefficient was 0.641. Conclusions:The scale of self-management of patients with dry eye has good reliability and validity and can be used in the assessment of dry eye patients to provide a reference basis for targeted guidance and intervention.

Objective:To analyze the influencing factors of genotypic drug resistance mutations in people living with human immunodeficiency virus and acquired immunodeficiency syndrome(PLWHA) who failed anti-retroviral therapy (ART) in Henan Province, in order to provide a basis for adjusting ART regimens and reducing drug resistance.Methods:PLWHA with virological failure (human immunodeficiency virus (HIV) RNA≥500 copies/mL) after receiving ART for more than 24 weeks were included in Henan Province from January 2018 to December 2022. Baseline CD4 + T lymphocyte counts, ART regimens and other clinical data were collected. HIV-1 gene subtypes and their drug resistance sequence mutations were detected in the Sixth People′s Hospital of Zhengzhou, and the sequences were submitted to the HIV Drug Resistance Interpretation System of Stanford University for comparison of test results. Genotypic drug resistance to nucleotide reverse transcriptase inhibitors (NRTI), non-nucleoside reverse transcriptase inhibitors (NNRTI), protease inhibitors (PI) and integrase inhibitors (INSTI) was determined. Multivariate logistic regression was used to analyze the influencing factors of drug resistance in patients with ART failure. Results:Among 982 PLWHA, the sequences of 899 cases were successfully amplified, and drug resistance was detected in 737 cases, with the drug resistance rate of 81.98%(737/899). The rates of resistance to NRTIs, NNRTIs, PIs and INSTIs were 71.97%(647/899), 79.31%(713/899), 5.23%(47/899) and 2.72%(20/734), respectively.The largest number of those who developed concomitant resistance to two classes of drugs was 588 cases (79.78%), mainly NRTI and NNRTI concomitant resistance in 583 cases (79.10%). There were 99 cases (13.43%) who developed resistance to only one class of drugs, and those who developed concurrent resistance to three classes of drugs were 48 cases (6.51%), and two cases (0.27%) were found to be resistant to all four classes of drugs mentioned above. A total of 10 HIV genotypes were detected, among which subtype B accounted for the most (59.73%(537/899)), followed by circulating recombinant form (CRF)01_AE subtype (21.91%(197/899)) and CRF07_BC subtype (9.45%(85/899)). The risk factors affecting the development of drug resistance were baseline CD4 + T lymphocyte counts, ART regimens and HIV-1 genotypes. The risk of drug resistance in patients with baseline CD4 + T lymphocyte counts <100/μL was 4.55 times (95% confidence interval ( CI) 2.69 to 7.70) higher than patients with CD4 + T lymphocyte counts≥250/μL, the risk of drug resistance in patients using 2NRTIs+ NNRTI regimen was 4.51 times (95% CI 1.75 to 11.63) higer than those using 2NRTIs+ INSTI regimen, and patients infected with B and CRF01_AE subtype was 2.18 times (95% CI 1.10 to 4.29) and 2.70 times (95% CI 1.26 to 5.78) higer than those with CRF07_BC subtype, respectively. Conclusions:The incidence of genotypic drug resistance in PLWHA with ART failure in Henan Province is high. Low baseline CD4 + T lymphocyte counts, 2NRTIs+ NNRTI regimens, and genotype B and CRF01_AE are risk factors for drug resistance in PLWHA.

Five new saponins, including three steroid saponins, paristenoids A-C (1-3), and two triterpenoid saponins, paristenoids D-E (4-5), along with four known ones (6-9) were isolated from the rhizomes of Paris polyphylla var. stenophylla. The structures of the isolated compounds were identified mainly by detailed spectroscopic analysis, including extensive 1D and 2D NMR, MS, as well as chemical methods. Compound 3 is a new cyclocholestanol-type steroidal saponin with a rare 6/6/6/5/5 fused-rings cholestanol skeleton, and this skeleton has been first found from the genus Paris. The cytotoxicities of the isolated compounds against three human three glioma cell lines (U87MG, U251MG and SHG44) were evaluated, and compound 7 displayed certain inhibitory effect with IC₅₀ values of 15.22 ± 1.73, 18.87 ± 1.81 and 17.64 ± 1.69 μmol·L^-1, respectively.
Humans ; Rhizome/chemistry* ; Steroids/chemistry* ; Liliaceae/chemistry* ; Saponins/chemistry* ; Triterpenes/analysis*

Humans ; Mutation/genetics* ; HIV Infections/drug therapy* ; Drug Resistance ; Treatment Failure ; China ; Drug Resistance, Viral/genetics*

Objective:To investigate the influencing factors of recurrence after radical resection of middle and low rectal cancer, and to establish a prediction model based on magnetic resonance imaging (MRI) measurement of perirectal fat content and investigate its application value.Methods:The retrospective cohort study was constructed. The clinicopathological data of 254 patients with middle and low rectal cancer who were admitted to Tianjin Union Medical Center from December 2016 to December 2021 were collected. There were 188 males and 66 females, aged (61±9)years. All patients underwent radical resection of rectal cancer and routine pelvic MRI examina-tion. Observation indicators: (1) follow-up and quantitative measurement of perirectal fat content; (2) factors influencing tumor recurrence after radical resection of middle and low rectal cancer; (3) construction and evaluation of the nomogram prediction model of tumor recurrence after radical resection of middle and low rectal cancer. Measurement data with normal distribution were represented as Mean± SD, and measurement data with skewed distribution were represented as M(rang) and M( Q1, Q2). Count data were described as absolute numbers. Univariate and multivariate analyses were conducted using the COX regression model. The rms software package (4.1.3 version) was used to construct the nomogram and calibration curve. The survival software package (4.1.3 version) was used to calculate the C-index. The ggDCA software package (4.1.3 version) was used for decision curve analysis. Results:(1) Follow-up and quantitative measurement of perirectal fat content. All 254 patients were followed up for 41.0(range, 1.0?59.0)months after surgery. During the follow-up period, there were 81 patients undergoing tumor recurrence with the time to tumor recurrence as 15.0(range, 1.0?43.0)months, and there were 173 patients without tumor recurrence. The preoperative rectal mesangial fascia envelope volume, preoperative rectal mesangial fat area, preoperative rectal posterior mesangial thickness were 159.1(68.6,266.5)cm3, 17.0(5.1,34.4)cm2, 1.2(0.4,3.2)cm in the 81 patients with tumor recurrence, and 178.5(100.1,310.1)cm3, 19.8(5.3,40.2)cm2 and 1.6(0.3,3.7)cm in the 173 patients without tumor recurrence. (2) Factors influencing tumor recurrence after radical resection of middle and low rectal cancer. Results of multivariate analysis showed that poorly differentiated tumor, tumor pathological N staging as N1?N2 stage, rectal posterior mesangial thickness ≤1.43 cm, magnetic resonance extra mural vascular invasion, tumor invasion surrounding structures were independent risk factors of tumor recurrence after radical resection of middle and low rectal cancer ( hazard ratio=1.64, 2.20, 3.19, 1.69, 4.20, 95% confidence interval as 1.03?2.61, 1.29?3.74, 1.78?5.71, 1.02?2.81, 2.05?8.63, P<0.05). (3) Construction and evaluation of the nomogram prediction model of tumor recurrence after radical resection of middle and low rectal cancer. Based on the results of multivariate analysis, the tumor differentiation, tumor pathological N staging, rectal posterior mesangial thickness, magnetic resonance extra mural vascular invasion, tumor invasion surrounding structures were included to construct the nomogram predic-tion model of tumor recurrence after radical resection of middle and low rectal cancer. The total score of these index in the nomogram prediction model corresponded to the probability of post-operative tumor recurrence. The C-index of the nomogram was 0.80, indicating that the prediction model with good prediction accuracy. Results of calibration curve showed that the nomogram prediction model with good prediction ability. Results of decision curve showed that the prediction probability threshold range was wide when the nomogram prediction model had obvious net benefit rate, and the model had good clinical practicability. Conclusions:Poorly differentiated tumor, tumor pathological N staging as N1?N2 stage, rectal posterior mesangial thickness ≤1.43 cm, magnetic resonance extra mural vascular invasion, tumor invasion surrounding structures are independent risk factors of tumor recurrence after radical resection of middle and low rectal cancer. Nomogram prediction model based on MRI measurement of perirectal fat content can effectively predict the probability of postoperative tumor recurrence.

Objective:To analyze the drug resistance characteristics of HIV/AIDS patients in Henan Province with antiretroviral treatment (ART) failure through the genotypic drug resistance detection.Methods:Blood samples were collected from HIV/AIDS patients who received ART for more than 6 months with viral loads ≥1 000 copies/ml in 18 cities of Henan from January 2018 to May 2021. The genotypic drug resistance detection was conducted by using an In-house drug resistance detection method. The drug resistance mutation (DRM) and antiretroviral susceptibility were analyzed by submitting the determined sequences to the Stanford HIV-1 drug resistance database. The information about patients' demographic characteristics and antiviral treatment data were collected.Results:A total of 887 HIV/AIDS patients with ART failure, 812 sequences were successfully amplified with the success rate of 91.54%. In the 812 patients, 676 were drug resistant (83.25%, 676/812). The drug resistance ratesto nucleoside reverse transcriptase inhibitors (NRTIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs), protease inhibitors (PIs), and integrase strand transfer inhibitors (INSTIs) were 73.40% (596/812), 80.54% (654/812), 5.54% (45/812), and 2.56% (17/663), respectively. There were significant differences in drug resistance rates among four types of drugs ( χ2=1 686.34, P<0.001). The drug resistance rate to two drugs was 66.38% (539/812), and the drug resistance rate to three drugs was 5.79% (47/812). A total of 9 subtypes of HIV-1were detected, in which subtype B accounted for 59.61%(484/812), followed by subtype CRF01_AE (22.17%, 180/812) and subtype CRF07_BC (9.48%, 77/812). There were significant differences in drug resistance rate among different subtypes ( χ2=21.33, P=0.001). Among NRTIs related mutation sites, the DRM rate of M184V/I was highest (63.42%, 515/812), followed by K65R (27.46%, 223/812). The top three DRM rates were detected for K103N/S (34.98%, 284/812), G190A/S (26.11%, 212/812) and V106M/I (24.63%, 200/812) among NNRTIs related mutation sites, and M46I (4.31%, 35/812), V82A/F (3.82%, 31/812), and I54V/MV (3.69%, 30/812) among PIs related mutation sites. While among INSTIs related mutation sites, E157Q/EQ had the highest DRM rate (3.47%, 23/663), followed by R263K (0.75%, 5/663) and G140A (0.75%, 5/663). The resistance to lamivudine and emtricitabine of NRTIs was at high-level (65.52%, 532/812), and the resistance to nevirapine (77.46%, 629/812) and efavirenz (71.18%, 578/812) of NNRTIs was also at high-level. The medium/high-level resistance to lopinavir/ritonavir of PIs was only 4.19% (34/812), the medium/high-level resistance to elvitegravir and raltegravir of INSTIs was 1.66% (11/663) and 1.21% (8/663), respectively, and no high-level resistance to bictegravir or dolutegravir was found. Conclusions:The drug resistance in HIV/AIDS patients with ART failure was high in Henan, characterized by high drug resistance rates to NRTIs and NNRTIs, and diverse and complex resistance mutations. So high resistance barrier ART-regimens were recommended, and the viral load monitoring and drug resistance testing after ART should be strengthened.

Objective : To analyze the application value of metagenomic next-generation sequencing (mNGS) in the detection of pathogenic bacteria in pulmonary infection ，and to provide a theoretical basis for clinical diagnosis. Methods : A retrospective study was conducted on 161 patients with suspected pulmonary infection，and samples of bronchoalveolar lavage fluid ，sputum and lung tissue were collected specimens ，pleural effusion and blood were simultaneously subjected to routine culture，respiratory pathogen nucleic acid detection and pathogenic microorgan- ism mNGS detection．The sensitivity ，specificity，positive predictive value ，and negative predictive value of the three detection methods were calculated respectively to evaluate the clinical application value of mNGS. Results: Among 161 patients with suspected pulmonary infection，113 cases were finally confirmed as pulmonary infection. Among the 161 patients enrolled，the sensitivity and negative predictive value of mNGS were improved compared with routine culture and nucleic acid detection of respiratory pathogens，and the difference was statistically signifi- cant(P＜0. 001) ．Although the specificity of mNGS was higher than that of respiratory pathogen nucleic acid de- tection，the difference was not statistically significant．The positive predictive value of mNGS was the highest，and was higher than that of respiratory pathogen nucleic acid detection，but had no statistical significance compared with conventional culture． Conclusion The sensitivity of mNGS in the diagnosis of pathogenic microorganisms in pul- monary infection is higher than that of conventional culture and nucleic acid detection of respiratory pathogens， which reduces the false negative rate of diagnosis and has a better comprehensive evaluation ; conventional culture detection has low sensitivity，but high specificity ; respiratory pathogen nucleic acid detection compared with con- ventional culture，the sensitivity is improved，but the specificity decreased.

Leishmaniasis is a prevalent cause of death and animal morbidity in underdeveloped countries of endemic area. However, there is few vaccine and effective drugs. Antimicrobial peptides are involved in the innate immune response in many organisms and are being developed as novel drugs against parasitic infections. In the present study, we synthesized a 5-amino acid peptide REDLK, which mutated the C-terminus of Pseudomonas exotoxin, to identify its effect on the Leishmania tarentolae. Promastigotes were incubated with different concentration of REDLK peptide, and the viability of parasite was assessed using MTT and Trypan blue dye. Morphologic damage of Leishmania was analyzed by light and electron microscopy. Cellular apoptosis was observed using the annexin V-FITC/PI apoptosis detection kit, mitochondrial membrane potential assay kit and flow cytometry. Our results showed that Leishmania tarentolae was susceptible to REDLK in a dose-dependent manner, disrupt the surface membrane integrity and caused parasite apoptosis. In our study, we demonstrated the leishmanicidal activity of an antimicrobial peptide REDLK from Pseudomonas aeruginosa against Leishmania tarentolae in vitro and present a foundation for further research of anti-leishmanial drugs.