ObjectiveTo investigate the effect of Zuoguiwan on the ovarian function in the rat model of cyclophosphamide-induced premature ovarian failure （POF） based on the changes of ferroptosis pathway. MethodForty SD rats were randomized into blank， model， and low- and high-dose （2， 8 g·kg^-1， respectively） Zuoguiwan groups， with 10 rats in each group. The rats in the other groups except the normal group were intraperitoneally injected with CTX at a dose of 50 mg·kg^-1 on the first day and 8 mg·kg^-1 from the second day to the fifteenth day for the modeling of POF. After modeling， the rats were administrated with corresponding drugs or normal saline by gavage for four weeks. Hematoxylin-eosin staining was performed to observe the pathological changes in the ovarian tissue. The mitochondria of the ovarian tissue was observed by electron microscopy. The serum levels of follicle-stimulating hormone （FSH）， estradiol （E₂）， luteinizing hormone （LH）， anti-Mullerian hormone （AMH）， malondialdehyde （MDA）， catalase （CAT）， superoxide dismutase （SOD）， and iron ion were measured by biochemical methods and enzyme-linked immunosorbent assay. Western blot was employed to determine the protein levels of glutathione peroxidase 4 （GPX4）， solute carrier family 7 member 11 （SLC7A11）， ferritin heavy chain （FTH1）， and acyl-CoA synthetase long chain family member 4 （ACSL4）. ResultCompared with the blank group， the model group showcased significantly increased atretic follicles， atrophied， fragmented， and vacuolated mitochondria， and reduced， loose， and disordered cristae in mitochondria. Compared with the model group， high-dose Zuoguiwan increased mature follicles， the volume of mitochondria in the ovary， alleviated the vacuolation， and improved the number and arrangement of mitochondrial cristae. Compared with the blank group， the modeling elevated the levels of iron， MDA， FSH， and LH， up-regulated the expression of GPX4， SLC7A11， and FTH1 （P<0.05， P<0.01）， decreased the activities of SOD and CAT， lowered the levels of E₂ and AMH， and down-regulated the expression of ACSL4 （P<0.05， P<0.01）. Compared with the model group， drug interventions lowered the levels of iron， MDA， FSH， and LH， down-regulated the expression of GPX4， SLC7A11， and FTH1 （P<0.05， P<0.01）， increased the activity of CAT， elevated the levels of E₂ and AMH， and up-regulated the expression of ACSL4 （P<0.05， P<0.01）. ConclusionZuoguiwan may inhibit the occurrence of ferroptosis by regulating the SLC7A11/GPX4 axis， thereby improving the ovarian function of POF rats.

Objective:To compare the value of 68Ga-1, 4, 7, 10-tetraazacyclododecane-1, 4, 7, 10-tetraacetic acid- D-Phe1-Tyr3-Thr8-octreotide (DOTATATE) and 18F-fluorodeoxyglucose (FDG) PET/CT imaging in the detection of bone metastasis in neuroendocrine neoplasm (NEN). Methods:From January 2014 to July 2019, 29 NEN patients (19 males, 10 females, age: 35-76 years) with bone metastasis who underwent 68Ga-DOTATATE and 18F-FDG PET/CT imaging within one month in Peking University Cancer Hospital & Institute were retrospectively enrolled. Patients were divided into Ki-67≤20% and Ki-67>20% groups according to the tumor proliferation activity, and osteolysis, osteogenesis and no change groups according to the CT findings of bone metastases. The differences of the number and radioactive uptake (maximum standardized uptake value (SUV max) ratio of bone lesion to normal bone (SUV T/B)) of detected bone metastases between 68Ga-DOTATATE and 18F-FDG PET/CT imaging were analyzed. χ2 and Mann-Whitney U tests were used to analyze the data. Results:The sensitivity of 68Ga-DOTATATE and 18F-FDG PET/CT imaging were 75.9%(22/29) and 82.8% (24/29) respectively, and there was no significant difference between the two modalities ( χ2=0.42, P>0.05). The numbers of cases with bone lesions detected by 68Ga-DOTATATE PET/CT imaging in pelvis, spine, ribs, proximal limbs, sternoclavicular scapula and skull were all higher than those of 18F-FDG PET/CT imaging (23, 22, 20, 14, 14, 10 vs 12, 19, 13, 11, 10, 6, respectively). The 68Ga-DOTATATE PET/CT imaging was significantly superior to 18F-FDG imaging in detecting bone metastases (9(3, 36) and 3(0, 18)) and SUV T/B(11.10(3.35, 22.30) and 1.60(1.05, 2.70); U values: 281.000, 77.000, both P<0.001). 68Ga-DOTATATE PET/CT imaging found more bone lesions in well differentiated NEN (Ki-67≤20%) group (11(2, 38) and 2(0, 13)) and osteogenic bone metastasis group (31(3, 100) and 3(0, 31); U values: 105.500, 69.500, both P<0.05). SUV T/B of 68Ga-DOTATATE PET/CT imaging was significantly higher than 18F-FDG PET/CT imaging in all subgroups ( U values: 3.000-22.000, all P<0.05). Conclusion:The 68Ga-DOTATATE PET/CT imaging is superior to 18F-FDG PET/CT imaging in the detection of bone metastasis in NEN.

To study the feasibility of ArcCheck verification system in dosimetric verification for stereotactic radiotherapy (SRT) the stereotactic radiotherapy (SRT) with flattening filter free (FFF) model.
 Methods: A total of 76 cases under SRT treatment plans were introduced into ArcCheck phantom and recalculated. Threshold criteria was set as (3%, 3 mm, 10%) or (2%, 2 mm, 10%). The calculated dose distribution and the measured dose distribution of ArcCheck phantom were compared by means of distance to agree (DTA) and Gamma analysis method respectively.
 Results: Based on the threshold criteria (3%, 3 mm, 10%), the relative and absolute mean pass rates of SRT treatment plans by DTA and Gamma analysis were greater than 95%. Based on the threshold criteria (2%, 2 mm, 10%), the relative and absolute mean pass rates of SRT treatment plan by DTA and Gamma analysis were about 90%. The dose pass rate of Gamma analysis method was slightly higher than that of DTA analysis method (P<0.001).
 Conclusion: The ArcCheck verification system is a rapid and accurate method for SRT dose verification, and discrepancies are found in different analysis methods.
Feasibility Studies ; Humans ; Phantoms, Imaging ; Radiosurgery ; methods ; Radiotherapy Dosage ; Radiotherapy Planning, Computer-Assisted ; Radiotherapy, Intensity-Modulated

Objective To investigate the effects of chitosan(CS)/pcDNA3.1(+) CrmA on the expression of interleukin-13 (IL-1β3) converting enzyme (ICE) and IL-1β in chondrocytes.Methods Rabbit chondrocytes were isolated and cultured.Chondrocytes were treated with PBS,10 μg/ml CS/pcDNA3.1(+) and CS/pcDNA3.1 (+) CrmA respectively for 6 hours.Then 10 ng/ml IL-1β was added into the culture medium.After 48 hours,the messenger RNA and protein expression of ICE and IL-1β in chondrocytes were detected by using real time polymerase chain reaction and western blotting.Results In CS/pcDNA3.1 (+) CrmA treated group (0.52 ±0.09),the mRNA expression of ICE inchondrocytes was significantly inhibited compared withcorresponding samples of CS/pcDNA3.1 (+) group (0.84±0.11,t=4.42,P＜0.01) and PBS group (1.00±0.10,t=6.58,P＜0.01).ICE protein expression in CS/pcDNA3.1 (+) CrmA treated group (0.20±0.03) was markedly lower than CS/pcDNA3.1 (+) (0.37±0.05,t=4.85,P＜0.01) and PBS treated group (0.44±0.07,t=6.68,P＜0.01).There was no significant difference of ICE mRNA and protein expressions in chondrocytes between CS/pcDNA3.1(+) group and PBS group.Significant difference of IL-1β mRNA expression was found in the three groups.IL-1 β mRNA expression level was significantly lower in CS/pcDNA3.1(+) CrmA treated group (0.55± 0.08) than CS/pcDNA3.1(+) (0.69±0.06,t=3.50,P＜0.01) and PBS group (0.99±0.04,t=11.12,P＜0.01) of chondrocytes.IL-1β protein expression level of chondrocytesin CS/pcDNA3.1(+) CrmA group (0.230±0.020) was significantly lower than CS/pcDNA3.1 (+) (0.450±0.060,t=5.07,P＜0.01)and PBS groups (0.610±0.090,t=8.70,P＜0.01) of cells.Significant difference of IL-1β mRNA and protein expression between CS/pcDNA3.1 (+) and PBS group was also observed (t=7.61,P＜0.01;t=3.63,P＜0.01).Conclusion CrmA mediated by chitosan can significantly suppress the mRNA and protein expression of ICE,thus down regulat the expression of IL-1β,which may be one of the mechanisms of CS/pcDNA3.1 (+) CrmA in the treatment of osteoarthritic cartilage degeneration.

Objective:To establish a method for the microbial limit test of Jingzhi Guanxin tablets. Methods: According to the methods in the 2010 edition and 2015 edition of Chinese Pharmacopoeia, the test was performed respectively. Results:Jingzhi Guanxin tablets showed obvious inhibitory effect on Bacillus subtilis. The bacteria count could be carried out by the culture medium diluting methods in the 2010 edition. The total amount of aerobe could be detected by the membrane filtration method in the 2015 edition. The total combined molds and yeasts count could be performed by the plate count method and the specified microorganism could be tested with the routine method. Conclusion:The above methods can be used for the microbial limit test of Jingzhi Guanxin tablets.

Objective To prepare and evaluate 99Tcm radiolabeled Cetuximab (C225) for imaging of EGFR specific binding.Methods Cetuximab antibody was reduced by 2-iminothiophene (2-IT).The radiolabeling of IT-Cetuximab with 99Tcm(99Tcm-IT-Cetuximab) was analyzed by HPLC,and was tested for in vitro stability and molecular integrity.The human lung cancer line (A549)-bearing nude mouse model was prepared for biodistribution and tumor targeted study.Tumor uptake was also measured by in vivo γ imaging.Results The labeling efficiency was 93.15％.The radiochemical purity was 96.46％ after purification.The in vitro stability was good in 5％ HSA,in which the radiochemical purity maintained above 80％ at 4 ℃ for 24 h.99Tcm-IT-Cetuximab showed good specific binding to tumor with peak uptake of (3.417±0.769) ％ID/g after 4 h.The T/NT ratio of blood increased to 1.454±0.174 at 24 h.γ imaging of A549-bearing nude mice xenografts also showed high T/NT ratio.Conclusions 99Tcm-IT-Cetuximab molecular probe could be prepared with high radiolabeling yield and radiochemical purity.It has excellent in vitro and in vivo stability,and shows specific uptake in A549 tumor.

Objective To investigate the effects of chitosan/pCDNA3.1 (+) CrmA on matrix metalloproteinase (MMP)-1 and tissue inhibitor of metalloproteinase (TIMP)-1 expression of chondrocytes induced by interleukin-1β (IL-1β) in mRNA and protein levels.Methods Rabbit chondrocytes were isolated and cultured.Chondrocytes were treated with PBS,10 μg/ml chitosan (CS) and chitosan/ pCDNA3.1 (+)CrmA respectively for 6 hours.Then 10 ng/ml IL-1β was added into the culture medium.After 48 hours,real time polymerase chain reaction (real time PCR) and Western blot assay were used to examine the changes of MMP1 and TIMP-1 in mRNA and protein levels.Results In CS/pCDNA3.1 (+)CrmA treated group (0.44±0.04),the messenger RNA expression of MMP-1 in chondrocytes was significantly suppressed compared with corresponding samples of PBS treated group (1.00±0.05) and CS treated group (0.76±0.07),There was significant difference between three groups (F=106.93,P＜0.01).The MMP-1 protein expression of chondrocytes in CS/pCDNA3.1 (+)CrmA treated group (0.28±0.03) was lower than that of PBS treated group (0.73±0.06) and CS treated group (0.46±0.05)(F=59.66,P＜0.01).No significant difference of TIMP-1 expression among the three groups was observed in mRNA and protein levels.Conclusion CS/pCDNA3.1(+) CrmA can significantly inhibit the mRNA and protein expressions of MMP-1 of chondrocytes induced by IL-1β,which leads to up-regulation the ratio of TIMPs to MMPs of IL-1β induced chondrocytes.It may be a part of the mechanisms of the therapeutic effects of CS/pCDNA3.1 (+)CrmA on osteoarthritis.

Objective:To establish a Taqman fluorescence quantitative PCR for the detection of Salmonella in drugs. Methods:Based on Salmonella specific gene, a pair of primers and a probe were designed, and DNA of Salmonella was extracted for the detec-tion. Results:The method was much specific, and the detection limit was 160 cfu/ml. The detection rate was 100% for the artificially contaminated samples. Conclusion:Fluorescence quantitative PCR can be applied in the rapid detection of Salmonella in drugs.

Objective:To study the application of petrifilm colony count plate in the microbial detection for cosmetics. Methods:Three common microbials in 14 batches of cosmetics were respectively detected by the method described in hygienic standard for cos-metics and petrifilm colony count plate, and the results were compared. Results:The results shown by the two methods had no statisti-cally significant difference (P>0. 05). Conclusion: Petrifilm colony count plate with light weight, fast and easy operation exhibits significance in the microbial detection for cosmetics.

Objective To investigate the effect of tail vein injection of exogenous bone marrow mesenchymal stem cells (BMSCs) on proliferative,osteogenic and adipogenic potential of BMSCs in ovariectomized osteoporotic rats.Methods The animal model of osteoporosis (OP) was established 3 months after ovariectomy (bilateral ovarian resection)in healthy 10-week-old female SD rats with the same batch and weight.The healthy 8-week-old SD GFP rats were selected in the same way as the BMSCs source.The experiment was divided into BMSCs group and control group.The BMSCs (1×106) marked with GFP were injected into BMSCs group rats by tail vein injection.One week after injection,BMSCs were isolated,cultured and purified in vitro to the 3rd,4th passage in all groups.The MTT method was used to detect the growth curves of BMSCs.The oil red O dyeing and alizarin red staining (ARS) were used to compare the adipogenic and osteogenic potential between the two kinds of BMSCs,respectively.The RT-PCR was used to detect the expression of osteogenesis-related genes (Runx2,osteocalcin and osteopontin) and adipogenesis-related genes (PPARγ and LPL) in BMSCs.Results Compared with the control group,the growth curve of BMSCs in BMSCs group rose significantly; the calcified nodule increased significantly after osteogenic induction; the lipid droplet decreased significantly after adipogenic induction.RT-PCR results showed that the Runx2,osteocalcin and osteopontin mRNA expression increased,while the peroxisome PPARγ and LPL mRNA expression decreased in BMSCs group.Conclution By tail vein injection of exogenous BMSCs,the proliferative and osteogenic potential of BMSCs in ovariectomized osteoporotic rats is enhanced,while the adipogenic potential is reduced,which provides a possibility of stem cell therapy for osteoporosis.