BACKGROUND:At present,modern medical treatment has certain limitations on the treatment of knee osteoarthritis.Traditional Chinese medicine alkaloids can effectively prevent and treat knee osteoarthritis through various mechanisms. OBJECTIVE:To review the mechanism of alkaloids in traditional Chinese medicine in the prevention and treatment of knee osteoarthritis,providing a scientific basis for the clinical development of drugs for the treatment of knee osteoarthritis. METHODS:CNKI,WanFang,PubMed,Web of Science and Google Scholar were retrieved for relevant literature published from database inception to May 2024.The key words were"knee osteoarthritis""osteoarthritis""osteoclast""chondrocyte""alkaloids"in Chinese and English.Duplicates and obsolete non-referenced literature were excluded,and a total of 68 eligible papers were included for further review. RESULTS AND CONCLUSION:Although traditional Chinese medicine has a long history of treating knee osteoarthritis,only a small number of natural compounds are in the preclinical stage of research against knee osteoarthritis.Alkaloids have a greater potential for the prevention and treatment of knee osteoarthritis,among which,sophocarpidine,oxymatrine,sinomenine,and betaine have been shown to be effective in the prevention and treatment of knee osteoarthritis by modulating multiple signaling pathways.Alkaloids can delay the progression of knee osteoarthritis by inhibiting inflammatory response,exerting antioxidant response,inhibiting chondrocyte apoptosis,promoting chondrocyte proliferation,and inhibiting osteoclast formation and differentiation.

Objective To compare the lipid-lowering efficacy and safety of fixed-dose combination and free combination of rosuvastatin and ezetimibe in hypercholesterolemia patients who fail to achieve low-density lipoprotein cholesterol(LDL-C)goal with statin monotherapy.Methods A total of 45 hypercholesterolemia patients who switched from statin monotherapy to fixed-dose combination of rosuvastatin and ezetimibe after failing to achieve target LDL-C goal admitted at cardiological departments of First Affiliated Hospital of Sun Yat-sen University,Nanhai Fourth People's Hospital,Foshan First People's Hospital,and Guangdong Provincial People's Hospital between March and June 2024 were enrolled and served as the study group.Another 120 hyper-cholesterolemia patients who treated with free combination of rosuvastatin and ezetimibe were se-lected from Xiamen Regional Health Medical Big Data Platform with propensity score matching and served as control group.The LDL-C level,LDL-C reduction,and changes in TC,HDL-C and TG levels in 4-6 weeks after the medication switch,as well as the safety indicators(AST,ALT,CK,Cre and eGFR)were compared between the two groups.Results In 4-6 weeks after the medication switch,the patients in the study group exhibited a significant decrease in LDL-C level(1.70±0.44 mmol/L vs 2.12±0.87 mmol/L,P＜0.01),obvious LDL-C reduction[(43.17±16.11)％vs(29.14±29.13)％,P＜0.01]when compared to those of the control group.The LDL-C goal attainment rate was significantly higher in the study group than the control group(71.11％vs 45.00％,P=0.003).In addition,there were no statistical differences in the levels of HDL-C and TG and the reductions of HDL-C and TG between the two groups in 4-6 weeks after treatment(P＞0.05).The study group obtained notably lower TC level and TC reduction than the control group in the time(P＜/0.05,P＜0.01).After treatment,no statistical differences were observed between the two groups in terms of AST,ALT,CK,Cre and eGFR(P＞0.05).Conclusion Com-pared to free combination of rosuvastatin and ezetimibe,fixed-dose combination can further reduce LDL-C level in hypercholesterolemia patients who have not achieved LDL-C goal with statin monotherapy,with higher LDL-C goal attainment rate and good safety.

Objective:To investigate the predictive value of serum vascular endothelial growth factor (VEGF), squamous cell carcinoma-associated antigen (SCCAg) and miRNA let-7a in lymph node metastasis and postoperative recurrence in patients with laryngeal cancer.Methods:A total of 82 patients with laryngeal cancer in the Second Central Hospital of Baoding from November 2017 to October 2019 were selected as the research subjects, including 18 cases of lymph node metastasis (metastasis group) and 64 cases of non metastasis (non metastasis group). The blood routine was tested before operation, and the baseline data, serum VEGF, SCCAg and miRNA let-7a levels were compared between the two groups. Logistic regression was used to analyze the related influencing factors of lymph node metastasis in patients with laryngeal cancer. The correlation between serum VEGF, SCCAg, miRNA let-7a levels and clinicopathological characteristics was analyzed. The receiver operating characteristic (ROC)curve was used to analyze the value of each index and the combined diagnosis of lymph node metastasis in patients with laryngeal cancer. After 1 year of follow-up, the serum VEGF, SCCAg and miRNA let-7a levels of patients with or without recurrent laryngeal cancer were compared. ROC curve was used to evaluate the value of VEGF, SCCAg, and miRNA let-7a in predicting the recurrence of laryngeal cancer.Results:There were statistically significant differences in tumor node metastasis (TNM) stage, degree of infiltration, degree of differentiation, serum VEGF, SCCAg, and miRNA let-7a levels between the metastasis group and non metastasis group (all P<0.05). Serum VEGF, SCCAg, miRNA let-7a levels in patients with laryngeal cancer were related to TNM stage, degree of infiltration and degree of differentiation (all P<0.05). The combined diagnosis of serum VEGF, SCCAg and miRNA let-7a levels in the diagnosis of lymph node metastasis in patients with laryngeal cancer showed that the diagnostic sensitivity and specificity were 88.89% and 70.31%, respectively. The serum VEGF and SCCAg levels of patients with recurrence after operation were higher than those without recurrence, and the level of miRNA let-7a was lower than those without recurrence (all P<0.05). The sensitivity and specificity of combined serum VEGF, SCCAg and miRNA LET-7a levels in predicting postoperative recurrence of laryngeal cancer were 72.97% and 91.11%, respectively. Conclusions:VEGF, SCCAg, miRNA let-7a in patients with laryngeal cancer have a certain correlation with clinicopathological characteristics, which can assist in the diagnosis of lymph node metastasis and help clinical prediction of postoperative recurrence in patients with laryngeal cancer, and provide a reference for the formulation of clinical treatment plans.

Objective: To investigate the prognostic factors of hyperamylasemia following pancreaticoduodenectomy (PD) . Methods: Clinical data of 359 patients were collected prospectively who underwent PD by the same group at Changhai Hospital of Navy Medical University from January 2017 to June 2018.There were 212 males and 147 females.The median age was 63 years old (range: 23 to 82 years old) .According to whether the patient′s serum amylase was greater than 120 U/L at 0 or 1 day after surgery,the patients were divided into hyperamylasemia group and non-hyperamylasemia group. Univariate analysis and multivariate analysis were used to find out the prognostic factors of hyperamylasemia after PD. Results: Of the 359 patients, 238 cases (66.3%) developed hyperamylasemia.The incidence rate of clinically related pancreatic fistula (15.1% vs.2.5%, P<0.01) , grade B/C post pancreatectomy hemorrhage (8.8% vs. 2.5%, P<0.01) , and surgical site infection (9.2% vs. 3.3%, P=0.04) was significantly higher in the hyperamylasemia group.The severity of complications (CD grade≥Ⅲ: 11.3% vs.4.1%, P=0.023) and postoperative hospital stay (11 days vs. 9 days, P=0.001) were higher in the hyperamylasemia group.In the multivariate analysis, the main pancreatic duct diameter (MPD) ≤3 mm (OR=4.469, 95% CI: 2.563-7.793, P<0.01) , pathological type of disease (pancreatic cancer or pancreatitis) (OR=0.230, 95% CI: 0.122-0.436, P<0.01) and soft texture of pancreas (OR=3.297, 95%CI: 1.930-5.635, P<0.01) were independent prognostic factors for hyperamylasemia. Conclusions Post-PD hyperamylasemia increased the incidence and severity of postoperative complications after PD.MPD≤3 mm, soft texture of pancreas and pathological type of disease were independent prognostic factors of hyperamylasemia.

Objective: To study the epidemiology and antimicrobial resistance of Gram-negative bacterial bloodstream infections in children, and to guide the choice of antimicrobials and the control of nosocomial infection. Method: Clinical data, bacteriology and antimicrobial susceptibility test results were collected retrospectively in hospitalized children who were diagnosed with gram-negative bacterial bloodstream infections in Yuying Children′s Hospital of Wenzhou Medical University from January, 2010 to December, 2015. Result: A total of 399 cases (253 male and 146 female) were identified. The age ranged from 16 hours to 16 years (median age 10.1 months). The majority of cases were collected from division of neonatology (n=261, 65.4%), followed by 31 cases (7.8%) from pediatric intensive care unit and 29 cases (7.3%) from Gastroenterology Department; 275 cases (68.9%) had underlying diseases, mainly including preterm birth(n=172), neonatal respiratory distress syndrome(n=67) and newborn asphyxia(n=53). Eighty cases had received invasive procedures and 20 had surgical operation; 149 cases (37.3%) were community-acquired and 250 cases (62.7%) were hospital acquired. Fifty cases had complications, among those, 40 cases had septic shock, 32 cases had multiple organ dysfunction syndrome and 7 cases had disseminated intravascular coagulation; 288 cases were cured, 48 improved, 17 gave up treatment and discharged, and 46 died; totally 408 strains were isolated from 399 children, including Enterobacteriaceae (346, 84.8%), non-fermentative Gram-negative bacteria (49, 12.0%) and other gram-negative bacteria (13, 3.2%). The resistance rates of Escherichia coli (n=175) and Klebsiella pneumoniae (n=106) to carbapenems, β-lactams enzyme and its inhibitors, amikacin and cefoxitin were all lower than 10%. Totally 245 multi-drug resistant strains (60.1%) were isolated, including 225 strains of Enterobacteriaceae and 18 strains of non-fermentative Gram-negative bacteria (P<0.01) . Nine strains of Carbapenem-resistant Enterobacteriaceae were isolated, which were all sensitive to amikacin and the sensitive rates to fluoroquinolone reached 88.9%. Five strains which were detected sensitive to tigecycline were all sensitive. The proportion of Klebsiella sp in Gram-negative bacteria between 2013-2015 and 2010-2012 were 32.9% and 21.2%, respectively. The resistance rates of Escherichia coli and Klebsiella pneumoniae to β-lactams and its enzyme inhibitors and carbapenems had no significant change. Conclusion Gram-negative bacterial bloodstream infections occur more frequently in newborns. Most children had combined underlying diseases. Escherichia coli and Klebsiella pneumoniae are the most common pathogens. β-Lactams and its enzyme inhibitors and carbapenems are the empirical choice of antimicrobial therapy for severe Enterobacteriaceae bloodstream bacterial infections.

BACKGROUND:Human and rat ovarian granulosa cels in dominant folicles have the phenomenon of expressing stem cel characteristics. OBJECTIVE:To investigate the expression of stem cel-related factors in rat ovarian granulosa cels. METHODS: After the paraffin sections of rat ovarian tissue, immunohistochemical method was used to detect CD34, CD133, ABCG2/Bcrp1, Pou5f1/Oct-4 expressions. Granulosa cels culturedin vitro were harvested by folicular puncture method, and then the immunohistochemical method was used to detect the expression of FSHR receptor in order to identify the purity of granule cels. In the cultured granulosa cels, CD44 and C-Kit expressions were detected immunohistochemicaly, RT-PCR was used to detect ABCG2/Bcrp1, Pou5f1/Oct-4, Nanog gene expressions in ovarian tissue and granulosa cels. RESULTS AND CONCLUSION:Immunohistochemistry detection on paraffin sections showed that a part of ovarian granulosa cels expressed CD34, CD133, ABCG2/Bcrp1 and Pou5f1/Oct-4, and the expression of Pou5f1/Oct-4 protein gradualy increased in the development of ovarian folicles, significantly enhanced during the luteal phase, and then disappeared after the formation of corpus albicans, displaying a periodic expression characteristics. FSHR receptor positive identification rate of primary cels harvested by the foliclar puncture method was more than 95%. Granulosa cels culturedin vitrowere mainly long spindle-shaped or diamond, and some cels presented with aggregation growth and expressed CD44 and C-Kit. RT-PCR test results showed that there were no Nanog in the ovarian tissue and cultured granulosa cels, low expression of Pou5f1/Oct-4 in the ovarian tissue, strong expression of ABCG2/Bcrp1 in the ovarian tissue, weak expression of Pou5f1/Oct-4 in the cultured granulosa cels, and strong expression of ABCG2/Bcrp1 in the cultured granulosa cels. These findings suggest that a part of granulosa cels in the rat ovarian have the characteristics of stem cels.

In order to study T-bet function in mouse cells, a novel retroviral vector expressing mouse T-bet and reporter gene Thy1.1 was constructed. Retrovirus particles were then produced by transfection of the recombinant retroviral plasmid into a packaging cell line Platinum-E. The recombinant retrovirus played considerable infection ability. T-bet expression was then identified by FACS after infection of CD4+ primary T cells from T-bet knockout mouse with recombinant retrovirus. To determine if exogenous expressing T-bet has normal function, we checked the expression level of T-bet target gene, Ifng. IFN-y expression was upregulated in the T-bet knockout T cells infected with recombinant retrovirus. In conclusion, we successfully constructed an effective mouse T-bet recombinant retroviral vector.
Animals ; Cell Line ; Genetic Vectors ; Interferon-gamma ; metabolism ; Mice ; Mice, Knockout ; Recombinant Proteins ; biosynthesis ; Retroviridae ; T-Box Domain Proteins ; biosynthesis ; T-Lymphocytes ; metabolism ; Transfection

ObjectiveTo observe the effects of hypoxia inducible factor-1 alpha (HIF-1α)-targeting small interfering RNA(siRNA) on the expression of HIF-1α and vascular endothelial growth factor (VEGF) in HaCaT ceils under hypoxic conditions. MethodsHaCaT cells were cultured and divided into four groups, normal control group (without any treatment), hypoxia group (cultured under hypoxic conditions for 24 hours),liposome control group (transfected with liposome followed by hypoxic culture for 24 hours), RNA interference group (transfected with HIF-1α-targeting siRNA/liposome complexes followed by hypoxic culture for 24 hours). Fluorescence real-time quantitative PCR was utilized to determine HIF-1oα and VEGF mRNA expression in HaCaT cells, and Western blot to detect HIF-1α and VEGF protein expression. ResultsNo significant difference was observed in the mRNA expression of HIF-1α between the hypoxia group and normal control group(0.907 ± 0.032 vs. 0.878 ± 0.034, F =1.108, P ＞ 0.05), while the expression levels of VEGF mRNA,HIF-1α and VEGF proteins were significantly higher in the hypoxia group than in the normal control group (0.935 ± 0.032 vs. 0.652 ± 0.053, 0.813 ± 0.047 vs. 0.236 ± 0.014, 0.791 ± 0.030 vs. 0.316 ± 0.013, all P ＜0.05). A significant decline was noted in the mRNA and protein expression levels of VEGF (0.230 ± 0.044 vs.0.978 ± 0.030, 0.213 ± 0.026 vs. 0.817 ± 0.049, both P ＜ 0.05) and HIF-1α(0.497 ± 0.033 vs. 0.806 ±0.040, 0.249 ± 0.028 vs. 0.833 ± 0.052, both P ＜ 0.05) in the RNA interference group than in the liposome control group. ConclusionsHypoxia may enhance the expression of HIF-1α and VEGF in HaCaT cells, and to inhibit the HIF-1α expression may suppress the expression of VEGF in HaCaT cells under hypoxia.

OBJECTIVE: To observe the effect of hypoxia inducible factor -1α (HIF-1α) small interfering RNA (siRNA) on the expression of HIF-1α and inducible nitric oxide synthase (iNOS) in HaCaT cells under hypoxia. METHODS: HaCaT cells were divided into 4 groups: the normal control group (without any treatment), the hypoxia group (under hypoxia for 24 h), the liposome control group (HaCaT cells transfected with liposome before hypoxia treatment), the RNA interference group (HaCaT cells transfected with siRNA sequences then under hypoxia for 24 h). Real-time PCR and Western blot were utilized to determine HIF-1α and iNOS mRNA and protein expression in HaCaT cells. RESULTS: There was no significant difference of the mRNA expression of HIF-1α between the hypoxia group and the normoxia group (P>0.05), but the protein expressions of HIF-1α was increased in the hypoxic group than that in the normoxia group (P<0.05). Both the mRNA and protein expression of iNOS were increased in hypoxic conditions than that in the normoxia (P<0.05). Decreases were more significant in the mRNA and protein expression of HIF-1α and iNOS in the RNA interference group than that in the liposome control group in HaCaT cells (P<0.05). CONCLUSION Hypoxia increased HIF-1α and iNOS expression in HaCaT cells and inhibition of HIF-1α expression decreased iNOS expression.
Cell Hypoxia ; Cell Line ; Humans ; Hypoxia-Inducible Factor 1, alpha Subunit ; genetics ; metabolism ; Keratinocytes ; cytology ; metabolism ; Nitric Oxide Synthase Type II ; genetics ; metabolism ; RNA Interference ; RNA, Messenger ; genetics ; metabolism ; RNA, Small Interfering ; genetics

Objective To investigate the changes in skin microcirculatory perfusion during induction of general anesthesia and the effects oftwo fluid therapy regimens in patients undergoing abdominal surgery.Methods Thirty-six ASA Ⅰ or Ⅱ patients aged 18-64 yr scheduled for elective major abdominal surgery were randomized to receive either 6% hydroxyetlayl starch(130/0.4)7 ml/kg(HES group,n=18)or lactated Ringer's solution 7 ml/kg(RL group,n=18)for compensatory intravascular volultne expansion(CVE)before tracheal intubation.Meanwhile both groups received continuous intravenous infusion of RL at a of 8 ml·kg~(-1)·h~(-1).Tracheal intubation was performed at 40 min after the onset of infusion.Anesthesia was maintained with with sevoflurane,remifentanil and rocuronium.Operation was started at 20 min after tracheal intubation.The microcirculatory perfusion was measured on forehead skin by using Doppler perfusion imaging system(LDPI)PI)at the onset of fluid infusion(T_0,baseline),the end of endotracheal intubation(T_1)and the onset of skin incision(T_2).Rwsults The MAP,HR,blood gases and body temperature were within the normal during the experiment and there was no significant difference between the 2 groups.The skin microcirculatory perfusion and CVP at T_1 were significantly higher in group HES than in group RL(P<0.05 or 0.01).Compared with the baseline value at T_0,the skin microcirculatory perfusion at T_1 was significantly increased in group HES(P<0.01),but there was no significant change in the skin microcirculatory perfusion at T_1 in group RL(P>0.05),the skin microcirculatory perfusion at T_2 was singificantly decreased in both groups(P<0.01),and CVP and PaO_2/FiO_2 at T_(1.2) were significantly increased,while Hb at T_(1.2) was significantly decreased in both groups(P<0.05).The skin microcirculatory perfusion in both groups was significantly lower at T_2 than at T_1(P<0.01).Conclusion The infusion of 6% HES 130/0.4 can improve the skin microcirculatory perfusion and the effect is better than that of RL during induction of general anesthesia in patients seheduled for abdominal surgery.