Objective To investigate the effect of the new polyamine analogue tetrabutyl propanediamine (TBP) on cell proliferation and the underlying mechanism. Methods MTT assay was performed to determine cell proliferation. Flow cytometry was performed to detect cell cycle transition. DNA fragmentation and mitochondrial membrane potential determinations were performed to detect cell apoptosis. The activity of key enzymes in polyamine catabolism was detected by chemiluminescence assay. Results TBP could significantly inhibit the proliferation of HL-60 cells by blockingcell cycle transition and by inducing apoptosis. The TBP-induced apoptosis of HL-60 cells was in a dose-dependent manner. The enzyme activities of SMO and APAO were also significantly increased in HL-60 cells after treatment with 100 μM TBP for 24 hours. Conclusions TBP, as a new putrescine analogue, could inhibit proliferation of HL-60 cells by increasing the enzyme activity of SMO and APAO and inducing apoptosis.

Abstract: Objective　To evaluate the value of bronchoalveolar lavage fluid Aspergillus-specific fluorescent PCR assay combined with galactomannan (GM) assay in the diagnosis of patients with non-neutropenic invasive pulmonary aspergillosis (IPA). Methods From March 2022 to December 2023, 113 hospitalized patients with clinically suspected IPA were selected from the Zibo First Hospital of Zibo City, Shandong Province. Bronchoalveolar lavage fluid samples from each patient were simultaneously subjected to potassium hydroxide microscopy, fungal culture, GM assay, and Aspergillus-specific fluorescence PCR assay. According to the diagnostic criteria of IPA, patients were divided into clinically diagnosed IPA and non-IPA groups. The values of these four methods for the diagnosis of IPA were compared. Results According to the diagnostic criteria for IPA, 37 out of the 113 suspected patients were clinically diagnosed as IPA. The proportion of diabetic patients was significantly higher in the IPA group compared to the non-IPA group (χ2=7.494, P=0.006); similarly, the proportion of patients using glucocorticoids was significantly higher in the IPA group (χ2=6.981, P=0.008). Patients in the IPA group more frequently showed cavitation within consolidation areas on imaging, which was statistically significant (χ2=15.603, P<0.001). There were significant differences in the sensitivity of the four fungal detection methods in the diagnosis of IPA (χ2=45.803, P<0.001), with Aspergillus-specific PCR assay showing the highest sensitivity at 94.59%. Specificity also varied significantly across the four methods (χ2=31.511, P<0.001), with the highest specificity being seen in potassium hydroxide microscopy and fungal culture at 100.00%. There were significant differences in the clinical coincidence rate of the four methods in the diagnosis of IPA (χ2=11.768, P=0.008), with Aspergillus-specific fluorescence PCR assay having the highest coincidence rate at 90.27%. The AUC of the ROC curve of Aspergillus-specific fluorescent PCR assay combined with the GM assay was 0.976 7, higher than 0.913 8 by Aspergillus-specific fluorescent PCR assay merely. Conclusions The combination of Aspergillus-specific fluorescent PCR assay and GM assay using bronchoalveolar lavage fluid could significantly improve the accuracy of IPA diagnosis in patients without neutropenia.

AIMTo establish the RP-HPLC fingerprint analysis for the quality control of Radix Angelicae dahuricae.

METHODSHPLC fingerprint analysis method of Radix Angelicae dahuricae was developed. Kromasil C18 column (250 mm x 4.6 mm ID, 5 microm) was used, with mixture of acetonitrile and water as mobile phase in a gradient mode. The flow rate was 1.0 mL x min(-1). The wavelength of measurement was 254 nm. Twenty-one batches of Radix Angelicae dahuricae were determined.

RESULTSThe 21 samples were classified as 4 clusters by cluster analysis and the 11 superior in producing area samples were confirmed to establish the mutual model. The samples' quality was assessed by Similarity Evaluation System for Chromatographic Fingerprint of TCM 2004.

CONCLUSIONThe method can be used to identify and evaluate the quality of Radix Angelicae dahuricae conveniently.

Angelica ; chemistry ; Chromatography, High Pressure Liquid ; methods ; Cluster Analysis ; Plant Extracts ; analysis ; chemistry ; standards ; Plant Roots ; chemistry ; Plants, Medicinal ; chemistry ; Quality Control ; Reproducibility of Results

Objective To investigate the correlation between ankle-brachial index and ultrasonography in lower ex- tremities vascular in type 2 diabetic patients.Methods 218 type 2 diabetic patients were enrolled,and ankle-brachial index(ABI)measurements and uhrasonography in lower extremities vascular were performed in all patients.Lower extremi- ties vascular disease(LEVD)were divided into six species according to the uhrasonography results and scored in accord- ante with its serious degree,then investigate the correlation between ABI and uhrasonography results score.All patients were divided into deferent groups according to uhrasonography results and ABI range,and then compared their deference. Results The lower the ABI,the more serious of the LEVD showed by ultrasonography.ABI and ultrasonic inspection score was significantly negatively correlated(r=-0.65,P

AIM:To investigate the effect of insulin on vascular smooth muscle cells (VSMCs) proliferation and to evaluate the intracellular signaling pathways involved.METHODS:VSMCs separated from Sprague-Dawley rats were used in this study. The proliferation of VSMCs induced by insulin was assayed by [3H]-thymidin incorporation. The protein expression and activity of p-ERK1/2 were determined by immunblot and [?-32P]ATP incorporation.RESULTS:Insulin induced cell proliferation in a concentration-dependent manner. The proliferative effect of insulin on VSMCs was inhibited partly by LY294002 (48.8%),an inhibitor of PI-3 kinase,and the ERK1/2 inhibitor PD98059 (43.6%),respectively. Moreover,phosphorylation of ERK1/2 and activity of ERK1/2 induced by insulin were also inhibited partly by LY294002.CONCLUSION:PI-3 kinase and ERK1/2 are involved in insulin induced VSMCs proliferation.

Objective To find out related proteins that could possibly predict curative effects of neoadjuvant chemotherapy(NACT) by applying surface-enhanced laser desorption/ionization time-of-flight mass spectrometry(SELDI-TOF MS)to detect the difference in protein spectrum of breast cancer tissues.Methods Cancer tissue samples from 30 cases of breast cancer patients were detected by SELDI-TOF MS before NACT to get protein spectrum of breast cancer tissues.After NACT,the curative effects were evaluated according to RECIST standard and patients were divided into effective group(19 cases with complete response or partial response)and invalid group (11 cases with stable disease or progressive disease).Results 11 different protein peaks (P ＜ 0.05) were screened out:3491 Da(t =3.189 P =0.004),5158 Da(t =3.897 P =0.001),5360 Da(t =2.157 P =0.04),7549 Da(t =2.173 P=0.038),8451 Da(t =2.258 P =0.032),8694 Da(t =3.234 P =0.003),9089 Da (t =2.653 P =0.013),10528 Da(t =3.127 P =0.004),13445 Da(t =2.231 P =0.034),15118 Da(t =3.255 P =0.003),and 44065 Da(t =2.554 P =0.017).They were all expressed higher in effective group than in invalid group.The difference had statistical significance(P ＜ 0.05).Conclusion SELDI-TOF MS technology can screen out protein spectrum sensitive to NACT,providing evidences for breast cancer patients whether or not taking NACT.

Objective To explore ideal surgical axillary management of early breast cancer,and to determine the feasibility of reducing false negative rate of sentinel lymph node biopsy(SLNB)by combination of axillary suspicious lymph node biopsy and SLNB.Methods From Jan.2008 to Oct.2009,42 consecutive cases with early breast cancer were enrolled.All patients underwent suspicious node hook-wire location by doppler ultrasonography before operation.SLNB and suspicious lymph node biopsy were performed during operation.Complete axillary nodes dissection(ALND)or level II dissection would be conducted according to the biopsy result.The difference of node status prediction between SLNB and SLNB with axillary suspicious lymph node biopsy and was compared.Results All the 42 cases successfully underwent SLNB (100%).There were 2 false negatives occurred in SLNB,resulting in false negative rate of 11%,sensitivity of 88.9%and accuracy of 95.2%in predicting axillary nodes status.By contrast,SLNB with axillary suspicious node biopsy showed a false-negative rate of 0%,sensitivity of 100%,and accuracy of 100%.Conclusions Compared to SLNB in early breast cancer,combination of suspicious node biopsy and SLAB has a tendency of reducing false negative rate.However,Because of the limited samples,the difference has no statistical significance(P=0.2500).

Objective To detect the sperm apoptosis rate and level of reactive oxygen species (ROS) in seminal plasma and explore their correlation among infertile males. Methods Ninety-two inferitile males were divided into varicocele (VC) group (n=32), leukocytospermia group(n=30) and the other cause group (n=30), and another 24 in vitro fertilization sperm samples were sereved as controls. The routine sperm parameters including seminal pH, sperm viability and sperm density were examined by computer assisted sperm analysis, the sperm apoptosis rate was asseseed using Annexin V/PI staining, and the ROS level in seminal plasma was detected by TBA method. The differences in seminal parameters between three infertile groups and control group were compared, and the correlation of sperm apoptosis rate with level of ROS in seminal plasma was explored in each group. Results The sperm viability of three infertile groups was significantly lower than that of control group (P<0.01). The sperm apoptosis rates and levels of ROS in seminal plasma in VC group and leukocytospermia group were significantly higher than those in control group (P < 0.05 or P < 0.01). The sperm apoptosis rate was positively correlated with the level of ROS in seminal plasma in leukocytospermia group(r=0. 573, P < 0.05). Conclusion The increased sperm apoptosis rate and level of seminal plasma ROS may be related to the infertility of patients with VC and leukocytospermia. The increased level of seminal plasma ROS may be one of the causes of increased sperm apoptosis rate in patients with leukocytospermia.

Objectives To investigate the pathogenic mechanism of herpes simplex encephalitis (HSE) and the mecha-nism of action of dexamethasone and acyclovir. Methods 102 male mice were randomly assigned to normal control group, HSV-1 infection group, acyclovir-treated group and combination-treated group. The model of HSE was established by in-tracranial injection of HSV-1 in mice except normal controls. One day after intracranial injection, mice in acyclovir-treated group and combination-treated group were intragastrically administrated with acyclovir, and mice in normal control group and HSV-1 infection group were intragastrically administrated with normal saline. Three days after intracranial injection, mice in combination-treated group were intraperitoneally injected with dexamethasone and mice in other groups were in-traperitoneally injected with normal saline. The neurological injury score and the expressions of IL-2 and IL-10 of the mice brain tissues in each group were compared at 3, 6 and 9 days after model establishment. Results The survival rate of mice was lowest in HSV-1 infection group and highest in combination-treated group (P<0.05). The neurological injury scores at 3, 6 and 9 days were highest in HSV-1 infection group and lowest in the combination-treated group (P<0.05). The changes in brain tissue pathology and HE staining were closely corresponded to the neurological injury scores. At 3, 6 and 9 days, the expression of IL-2 was increased at first and then decreased, but the expression of IL-10 was gradually increased. The expressions of IL-2 and IL-10 were highest in HSV-1 infection group and lowest in combination-treated group. And differ-ences were significant among all groups at each time point (P<0.05) except between combination-treated group and normal control group (P>0.05). Conclusions In comparison with acyclovir monotherapy, combined treatment with dexamethasone and acyclovir for HSE can reduce the expression of IL-2 and IL-10, relieve the clinical symptoms, and increase the survival rate.

A strain of pseudomonas sp. L-11 could produce biosurfactants by use of glucose as substrate. The fermentation conditions in shaking flask and 1L fennentor were investigated. The optimal medium and (he fermentation technology were demonstrated. In 1L fermentor, the interracial tension between the diluent of L-11 (10%) and crude oil is 5.3?10~3mN/m, which is fit for microbial enhanced oil recovery. The scale-up technology of L-11 was also studied.