OBJECTIVETo investigate the inhibitory effect of puerarin on invasive and metastatic abilities of tumor cells, and its possible mechanism through observing its impacts on the migratory, adhesive and invasive capacities of human oophoroma cells HO-8910 to the artificial recombined basement membrane.

METHODSExpression of estrogen receptor (ER) in HO-8910 cells was detected using PCR assay. Effects of puerarin on HO-8910 proliferation was detected with MTT assay; on its adhesion potential was tested with cell-matrigel adhesion assay, and on invasive and migratory capacities were measured with Transwell matrigel invasion assay and Transwell motility assay respectively.

RESULTSER was positively expressed in HO-8910 cells. After being treated with 20 micromol/L puerarin for 12 h, the adhesive test showed that OD value in the tested group was significantly lower than that in the control (P < 0.01), the inhibiting rate reached 50.63%; and the Transwell assay showed a significant lowering of penetrated cells (P < 0.01), the inhibition rate for invasion was 38.59% and that for motility migration 40.63%. The number of penetrated cells was lower in the group intervened with combination of Puerarin and estrogen than in the group intervened with estrogen alone, 33.40 +/- 3.30 vs 48.05 +/- 3. 56 for invasion and 35.35 +/- 3.03 vs 52.45 +/- 1.04 for motility (all P < 0.01).

CONCLUSIONPuerarin can inhibit the adhesion, invasion and migration of HO-8910 cells, plays an antagonist effect against the stimulation of estrogen on the malignant behavior of tumor cells.

Cell Adhesion ; drug effects ; Cell Line, Tumor ; Cell Movement ; drug effects ; Female ; Humans ; Isoflavones ; pharmacology ; Neoplasm Invasiveness ; Neoplasm Metastasis ; Ovarian Neoplasms ; pathology ; Receptors, Estrogen ; metabolism

Objective To observe the distribution of oligodexynucleotide (ODN)MT01 in main organ tissues of the rats at different time points and to discuss the regularity of the distribution of MT01 preliminarily. Methods 60 male Wistar rats were randomly divided into experimental group(n=30)and control group(n=30). The rats in experimental group was locally injected with Cy5 labeled MT01 in gingival mucosa,whereas the rats in control group were injected with MTO1.The samples of rat lung,liver spleen,kidney,heart,and brain tissues were collected at 15 min,1 h,4 h,8 h,16 h,1 d,2 d,3 d,4 d,and 5 d after injection,and the distribution of MT01 fluorescence was observed by laser scanning confocal microscope.The ratio of fluorescence positive cells indicated the amount of MT01 that had been taken up by different organs.Results No positive fluorescence cells were observed in control group.Whereas,in experimental group ,the positive fluorescence cells were detected in the tissue samples of lung,liver,spleen and kidney but not in the tissue samples of heart and brain.The positive fluorescence cells distributed focally in kidney tissue and presented primarily in the cytoplasm of renal tubular epithelial cells.The ratios of positive fluorescence cells changed regularly with time in liver, spleen and kidney tissues and the highest level was detected at 4,3 and 4 d after injection.No distinct regularity of the ratio of positive fluorescence cells was observed in lung tissue.Conclusion MT01 can be taken up by liver,spleen and lung tissue and primarily by kidney with regularity in distribution.

Objective: To monitor the adverse events following immunization (AEFI) with combined attenuated live measles, mumps and rubella vaccines (MMR) in Huzhou City from 2015 to 2021, so as to provide insights into the implementation of the MMR immunization strategy. Methods: All AEFI caused by MMR immunization in Huzhou City from 2015 to 2021 were captured from the AEFI Monitoring Information Management System of the Immunization Planning System of Chinese Disease Control and Prevention Information System, and the incidence, clinical features and epidemiological features of AEFI were analyzed descriptively. Results: The reported incidence of AEFI caused by MMR immunization appeared a tendency towards a rise in Huzhou City from 2015 to 2021 (χ2trend=124.126, P<0.001). Totally 324 386 doses of MMR vaccines were immunized, and 317 cases with AEFI were reported, with an reported incidence rate of 9.77/104 doses. Following two-dose immunization, the reported incidence of AEFI caused by two-dose MMR immunization was significantly lower than by one-dose immunization (6.01/104 doses vs. 25.43/104 doses; χ2=113.692, P<0.001). The incidence rates of general reactions, abnormal reactions and coincidental events were 6.20/104 doses, 3.42/104 dose and 0.15/104 doses, respectively. Fever and allergic rash were predominant clinical manifestations of AEFI, and no vaccine quality accident, inoculation accident or psychogenic reaction were reported. There were 246 (77.60%) cases with AEFI within 24 hours following vaccination, and among children with AEFI, there were 173 boys (54.57%), and 200 children (63.09%) age ages of less than one year (63.09%). AEFI was reported in each quarter, and 99 cases (31.23%) were reported in the fourth quarter. The largest number of children with AEFI was reported in Wuxing District (78 cases, 24.61%). Conclusions The safety of MMR vaccination is high in Huzhou City. General reaction is the predominant AEFI, which mainly occurs within 24 hours following vaccination. Two-dose MMR vaccination does not increase the risk of AEFI.

Objective: To investigate the incidence of adverse events following immunization (AEFI) with inactivated influenza vaccine among the elderly in Huzhou City, Zhejiang Province, so as to provide insights into safety monitoring and evaluation of inactivated influenza vaccines. Methods: Data pertaining to surveillance on AEFI with inactivated influenza vaccines among the elderly at ages of 60 years and older in Huzhou City from 2020 to 2022 were collected from the AEFI Monitoring Information Management System of the Immunization Planning System of Chinese Disease Control and Prevention Information System, including demographics, time of AEFI occurrence, classification of AEFI and clinical syndromes, and the reported incidence and epidemiological features of AEFI with inactivated influenza vaccines were analyzed using a descriptive epidemiological method. Results: Totally 84 elderly cases at ages of 60 years and older were reported with AEFI with inactivated influenza vaccines in Huzhou City from 2020 to 2022, with a reported incidence rate of 9.83/105 doses, and the reported incidence rates of AEFI with trivalent and quadrivalent inactivated influenza vaccines were 9.74/105 doses and 48.71/105 doses, respectively. The reported incidence rates of general, abnormal, coincidence and psychogenic reactions were 7.96/105 doses, 1.52/105 doses, 0.23/105 doses and 0.12/105 doses, respectively, and no vaccine quality accidents or wrong vaccine administered were reported. The cases with AEFI included 52 women and 32 men, and most cases were aged from 60 to 69 years (44 cases, 52.38%). The highest incidence of AEFI was reported in Nanxun District (17.94/105 doses), and there were 79 cases (94.05%) with AEFI within 24 hours following vaccination. The clinical symptoms mainly included fever, local redness and swelling, and local induration, with reported incidence rates of 2.22/105 doses, 3.74/105 doses, and 1.99/105 doses, respectively. Conclusions The reported incidence of AEFI with inactivated influenza vaccines is low among the elderly at ages of 60 years and older in Huzhou City, with general reactions as predominant AEFI, and most AEFI occurs within 24 hours following vaccination.

Background Femtosecond laser in situ keratomileusis (LASIK) inevitably injury keratocytes and corneal nerve fibers.The research report about postoperative morphological changes of corneal nerve regeneration and keratocytes in femtosecond LASIK is still rare.Objective The aim of this study was to observe the kinetic changes of keratocytes and corneal nerve in corneal flap after femtosecond LASIK.Method Femtosecond laser manufacture corneal flap of LASIK surgery was performed on 60 eyes of 30 patients with refractive error using both femtosecond laser system and excimer laser treatment system.The repair of corneal wound was examined by slit lamp microscope,and the morphology of keratocytes and corneal nerve were observed with confocal microscope 1 week,1month,3 months after surgery,respectively.Results No haze or flap folds were found under the slit lamp microscope from 1 week through 3 months after operation.One week after surgery,the corneal stromal cells at the interface of the corneal flap appeared to be a mild activation status in 42 eyes (70％),but the activated cells gradually reduced with lapse of time.Three months after surgery,mild activation state still was found in 7 eyes (12％).One week after surgery,independent,short (＜50 μm),curved subbasal nerve fibers were exhibited in 7 eyes (12％),and curved filamentous nerve fibers were discovered in 48 eyes (80％) one month after surgery.The nerve fiber length of subbasal nerve was ＞200 μm in 27 eyes (45％) and classes beaded structure appeared 3 months after operation but were still different with preoperative subbasal nerve fibers.One week after the operation,filaments or discontinuous nerve fibers could been seen in 46 eyes (77％) at theinterface,and long nerve fibers or filamentous nerves were visible around the terminal or periphery of nerve fibers in 49 eyes (82％) one month after surgery,and long nerve fibers or filaments of nerve fibers were visible in 57 eyes (95％) 3 months after the surgery.Conclusions Femtosecond LASIK cause wound reaction at cellular level.Corneal nerve fibers recover with the extension of time,but there are still some morphological differences 3 months after surgery from preoperation.

Objective To study the effect of body fat distribution on weight-losing treatment.Methods 471 simple obesity patients were divided into two groups: visceral fat normal group and visceral fat excess group, which is according to the visceral fat measured by Inbody720 Body Compositions Analysis. An organized program with energy intake controlling and exercise for one month was taken. Body weight of all patients was measured before and after the program.Results After weight-losing treatment, the losing weight of visceral fat normal group was significantly higher than visceral fat excess group (P＜0.01), and this difference is in existence in both gender (P＜0.05 of male, P＜0.01 of female). There were no gender difference in weight-losing (P＞0.05).The higher the BMI was, the more weight-losing was. There was no age difference (P＞0.05). Conclusions The effect of weight-losing is associated with visceral fat and BMI.

Objective To construct a recombinant oncolytic virus vvmIL33 that can steadily se-crete mouse IL-33 protein (mIL-33) in targeted tumor cells and to study its synergistic inhibitory effect on tumor. Methods Mouse IL-33 gene sequence was amplified by PCR and inserted into the eukaryotic ex-pression vector pCMS1. The constructed pCMS1-mIL33 was transfected into the parent virus (vJS6)-infected cells by Lipofactamine. Recombinant oncolytic virus vvmIL33 was purified by cell flow sorting. Enzyme-linked immunosorbent assay ( ELISA) was used to detect the level of mIL-33 protein in the culture superna-tant of vvmIL33-infected tumor cells. Recombinant oncolytic virus vvmIL33 and parental virus vJS6 were re-spectively used to infect tumor cells, and then analyzed by plaque formation assay and MTS kit. T cell co-culture experiments were performed to examine the anti-tumor ability of T cells induced by vvmIL33-infected tumor cells. Results Electrophoresis results of the recombinant plasmid pCMS1-mIL33 showed that mIL-33 gene was inserted successfully. Compared with the control group, vvmIL33 could steadily secrete high levels of mIL-33 protein in MC38 cells after infection (P<0. 001). Results of the plaque formation assay showed that vvmIL33-or vJS6-infected CV1 and MC38 cells produced similar amounts of virus at various time points without statistical difference (P>0. 05). Under different multiplicity of infection (MOI), the lytic ability of vvmIL33 against tumor cells was similar to that of vJS6 without statistical difference (P>0. 05). In the T cell co-culture experiments, the concentration of INF-γ protein produced by T cells in the vvmIL33-infected MC38 cell group was significantly increased as compared with that of the vJS6 group (P<0. 05). Moreover, the cytotoxic effect of induced T cells on tumor cells was also significantly increased (P<0. 05). Conclusion The recombinant oncolytic virus vvmIL33 was successfully constructed without damaging its ability to repli-cate and induce tumor cell lysis. Oncolytic virus carrying mIL-33 enhanced the immune effect of T cells and increased anti-tumor effect.

Objective To explore the correlation between polarization status of microglia/macrophages(MG/MP)in brain tissue and edema around hematoma in patients with acute cerebral hemorrhage.Methods A total of 52 patients with acute intracerebral hemorrhage admitted to Anyang People's Hospital from December 2020 to November 2022 were selected as the research subjects.All patients underwent craniotomy to remove hematoma,and the normal brain tissue in the cortical area that was not invaded by the hematoma and the fragmented brain tissue around the hematoma(brain tissue around the hematoma)on the surgical pathway were obtained.The expression levels of inflammatory factors such as interleukin(IL)-1β,IL-6,tumor necrosis factor-α(TNF-α),IL-10 and transforming growth factor-β(TGF-β)protein in brain tissue were detected by Western blot.The expression levels of IL-1 β,IL-6,TNF-α,IL-10 and TGF-β mRNA in brain tissue were detected by fluorescence quantitative polymerase chain reaction.The levels of M1-type and M2-type MG/MP in brain tissue was detected by immunofluorescence confocal technique.CT images data of patients before operation were collected and the relative-erihema-tomal edema(r-PHE)was calculated.The patients were divided into high r-PHE group(2.0≤ r-PHE＜2.5)and low r-PHE group(1.5＜r-PHE＜2.0)according to r-PHE.The relative expression of IL-1 β,IL-6,TNF-α,IL-10 and TGF-β mRNA in brain tissue around the hematoma of patients between the high r-PHE group and the low r-PHE group was compared.Results The relative expressions of IL-1 β,IL-6,TNF-α protein and mRNA in brain tissue around the hematoma were significantly higher than those in the normal brain tissues(P＜0.05),but there was no significant difference in the relative expressions of IL-10 and TGF-β protein and mRNA between the brain tissue around the hematoma and the normal brain tissue(P＞0.05).The levels of M1 type and M2 type MG/MP in the brain tissue around the hematoma were significantly higher than those in normal brain tissue(P＜0.05).The relative expressions of IL-1β,IL-6 and TNF-α mRNA in the brain tissue around the hematoma of patients in the high r-PHE group were significantly higher than those in the low r-PHE group(P＜0.05),and there was no significant difference in the relative expressions of TGF-β and IL-10 mRNA in the brain tissue around the hematoma of patients between the two groups(P＞0.05).Conclusion The levels of pro-inflammatory factors and M1-type MG/MP are increased in the brain tissue around the hematoma in patients with acute cerebral hemorrhage,and the degree of polarization of M1-type MG/MP is consistent with the degree of edema around hematoma after cerebral hemorrhage.

OBJECTIVETo establish a quantitative HPLC method for determination of protopine, palmatine hydrochloride, bicuculline and tetrahydropalmatine, in Corydalis decumbens.

METHODThe separation was performed on a ZORBAX Eclipse XDB-C18 column (4.6 mm x 150 mm, 5 microm) at a flow rate of 1.0 mL x min(-1) using mixtures of two solvents [A(20 mmol x L(-1) ammonium acetate)-B(acetonitrile)]: with a gradient elution. The column oven temperature was 30 degrees C and the detection wavelength was set at 280 nm.

RESULTThe 4 alkaloids were well separated by this HPLC method. Linearifies of protopine, palmatine hydrochloride, bicuculline and tetrahydropalmatine were good in the ranges of 1.44-46.0 (r = 0.999 4), 1.2640.2 (r = 0.999 8), 1.37-44.0 (r = 0.999 9), and 1.3643.6 mg x L(-1) (r = 0.999 9), respectively. The average recoveries were 98.2% with RSD 2.7% for protopine, 101.9% with RSD 2.5% for palmatine hydrochloride, 102.8% with RSD 3.5% for tetrahydropalmatine, and 98.8% with RSD 3.1% for tetrahydropalmatine.

CONCLUSIONThis method is proved to be convenient, reliable and accurate., and it can be used for quality control of C. decumbens.

Alkaloids ; analysis ; Chromatography, High Pressure Liquid ; Corydalis ; chemistry ; Drug Stability ; Plants, Medicinal ; chemistry ; Reproducibility of Results

OBJECTIVETo study the abnormal reactions of a series of free radicals and the oxidative damages induced by free radical abnormal reactions in the bodies of patients with chronic glomerulonephritis.

METHODSEighty chronic glomerulonephritis patients (CGNP) and eighty healthy adult volunteers (HAV) were enrolled in a random control study, in which concentrations of nitric oxide (NO) in plasma, lipoperoxides (LPO) in plasma and in erythrocytes, and vitamin C (VC), vitamin E (VE) and beta-carotene (beta-CAR) in plasma as well as activities of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX) in erythrocytes were determined with spectrophotometric assays.

RESULTSCompared with the average values of the above biochemical parameters in the HAV group, the average values of NO in plasma, and LPO in plasma and erythrocytes in the CGNP group were significantly increased (P = 0.0001), while those of VC, VE and beta-CAR in plasma as well as those of SOD, CAT and GPX in erythrocytes in the CGNP group were significantly decreased (P = 0.0001). Pearson product-moment correlation analysis showed that with increase of the concentration of blood creatinine as well as prolongation of the course of disease in the CGNP, the concentrations of NO in plasma, and LPO in plasma and erythrocytes in the CGNP increased gradually, while the concentrations of VC, VE and beta-CAR in plasma as well as the activities of SOD, CAT and GPX in erythrocytes in the CGNP decreased gradually (P = 0.002454-0.000001). The relative risk ratio (RR) of the above biochemical parameters reflecting oxidative damages in the bodies of CGNP ranged from 6.061 to 72.429. The reliability coefficient (alpha) that the above biochemical parameters were used to reflect the oxidative damages of the CGNP was 0.8137, standardized item alpha = 0.9728, Hotelling's T-Squared = 1135680.191, F = 53274.6478, P = 0.000001.

CONCLUSIONSThe findings in this study show that in the bodies of CGNP a series of free radical chain reactions result in severe pathological aggravation and induce oxidative damages in their bodies. Therefore, suitable dose of antioxidants should be supplemented to them so as to alleviate oxidative damages in their bodies.

Adult ; Antioxidants ; pharmacology ; therapeutic use ; Catalase ; pharmacology ; Chronic Disease ; Erythrocytes ; enzymology ; Female ; Free Radicals ; adverse effects ; Glomerulonephritis ; physiopathology ; Glutathione Peroxidase ; pharmacology ; Humans ; Lipid Peroxidation ; Male ; Nitric Oxide ; adverse effects ; analysis ; Oxidation-Reduction ; Oxidative Stress ; Superoxide Dismutase ; pharmacology