Objective To explore the synergistic effectiveness of hyperthermia and chemotherapy in the treatment of advanced gastric cancer.Methods Eighty-nine patients with advanced gastric cancer were randomly assigned to a study group which received a CapeOx chemotherapy regimen supplemented with hyperthermia or to a control group which received only the CapeOx regimen.The regimen consisted of capecitabine (1000 mg/m2,bid,orally for 14 consecutive days) plus oxalipaltin (130 mg/m2) on day 1.The hyperthermia was at 43℃ for 60 min in the tumor area on day 1 and twice a week thereafter.One cycle was 21 days.After 2 treatment cycles,efficacy was evaluated according to RECIST standards,improvements in the quality of life were assessed according to Karnofsky's performance status (KPS) and the side-effects of therapy were recorded.Results The response rate was 68.9％ in the study group and 36.4％ in the control group,showing a significant difference between the groups after two treatment cycles.The median progress-free survival (PFS) was 8.3 months in the study group vs 5.2 months for the controls.The 1-year survival rate was 66.4％ vs 45.5％ and the rate of improvement in KPS was 77.8％ vs 45.5％.All these differences were statistically significant.The common adverse effects were gastrointestinal toxicity,marrow depression and peripheral nerve abnormalities,but these adverse effects were all mild and similar in the two groups.Conclusion Hyperthermia when combined with the CapeOx chemotherapy regimen might improve the therapeutic effect in advanced gastric cancer without obviously increasing the adverse effects.

Aim TodevelopandvalidateaLC-MS/MS assay to quantify halometasone in rabbit plasma and study pharmacokinetics of halometasone after dermal topical administration of Halometasone Cream.Meth-ods Theplasmasamplewassubmittedtoliquid-liquid extraction using methyl tertiary butyl ether,with dexa-methasone as the internal standard (IS ).Chromato-graphic separations were performed on a Diamonsil C18 column(100 mm ×4. 6 mm,5 μm)with a linear gra-dient of methanol and 2 mmol · L-1 ammonium ace-tate.Halometasone and dexamethasone(IS)were ion-ized with an ESI source operated in negative ion mode, and the detected ions were m/z 503. 1→413. 0 (halo-metasone),m/z 391. 0→361. 0 (dexamethasone ). The test article could be monitored in rabbit plasma when following single dermal topical administration of Halometasone Cream at 1 g/100 cm2 to rabbits by u-singavalidatedLC-MS/MSassay.Results Calibra-tion curve was linear over the concentration range of0. 02~20 μg·L-1 in rabbit plasma.For low,medi-um,high concentration of QC solutions,the intra-and inter-day precision was in the range of 3. 72% ~7. 87%, and the accuracy was within 99. 1% to 103%. The pharmacokinetic parameters in rabbits were as follows:Tmax,Cmax,AUC0-t,T1/2 was (7. 38 ± 1. 06)h,(1. 16 ±0. 527)μg·L-1,(18. 8 ±7. 23)h·μg·L-1 ,(13. 8 ±3. 70)h,respectively.Conclusions ThisLC-MS/MSanalysismethodhashighsensitivi-ty,and sample processing method is simple,which has been rigorously validated.The method could be suc-cessfully applied to the pharmacokinetic study of halo-metasone after skin administration of Halometasone Cream to rabbits.

Objective To observe the inhibitory effect on lymph node metastasis of pancreatic cancer and lymphangiogenesis in mice by injection of KAll gene within xenograft tumor.Methods Pancreatic cancer cell line MiaPaCa-2 wag used to construct the nude mice models bearing tumors,then the mice were divided into normal saline group,Ad group and Ad-KAI1 group.Since the successful model construction,normal saline,Ad,Ad-KAI1 was injected every week for 3 times,respectively in the three groups,then the tumor size was documented.50 d after model construction,the tumor and enlarged lymph nodes were collected and subjected to pathological exam,and the expression of LYVE-1 and the MLVD in xenograft tumor was detected by immunohistochemistry.Results Two weeks after MiaPaCa-2 implantation,the model was 100％ successfully constructed.The growth curve of subcutaneous tumor among 3 groups was not statistically significant (P＞0.05) ; the weights of subcutaneous tumor in the 3 groups were (2514.4 ±351.3),(2466.1 ± 295.5),(2294.4±255.4) mg after 50 d,and the difference among the 3 groups was not statistically significant (P ＞0.05).Enlarged lymph nodes metastasis was observed in 8 mice (80％) in normal saline group,and 20 lymph nodes were collected,with 2.0 lymph nodes per mice; and enlarged lymph nodes metastasis was observed in 7 mice (70％) in Ad group,and 15 lymph nodes were collected,with 1.5 lymph nodes per mice; while enlarged lymph nodes metastasis was observed in 4 mice (40％) in Ad-KAI1 group,and 6 lymph nodes were collected,with 0.6 lymph nodes per mice.All the lymph nodes were confirmed to be metastasis of the primary tumor after pathologic exam.The difference of lymph nodes metastasis,number of lymph nodes metastasis per mice among the 3 groups was statistically significant (F =3.14,3.35,P ＜ 0.05).The MLVD of subcutaneous tumor among the 3 groups was (18.70 ± 5.60),(19.40 ± 4.58),(9.80 ±4.10)/400 times magnification,the MLVD of Ad-KAI1 group was significantly lower than those in normal saline group and Ad group (F10.76,11.36,P ＜ 0.05),but the difference between normal saline group and Ad group was not statistically significant.Conclusions KAI1 can inhibit the lymph node metastasis of pancreatic cancer,and the mechanism may be related with decreased lymphangiogenesis and reduced lymphatic vessel density.

Objective To compare the effect of autologous bone marrow stem cells transplantation on liver function between first and second transplantation in decompensated liver cirrhosis patients.MethodsA total of 45 decompensated liver cirrhosis patients were enrolled, and 23patients in first transplantation group were transplanted with autologous bone marrow stem cells through femoral artery when condition was stable after medical treatment.In second transplantation group, 22 patients were accepted second transplantation in 4-12 month after the first transplantation.All the patients undergone routine blood test, congulation test and liver function examination at the fourth week and eighth week after transplantation.ResultsEight weeks after transplantation, the liver function was improved obviously in both first and second autologous bone marrow stem cells transplantation.The level of albumin in patients of second transplantation group increased from (37.26± 5.90) g/L before transplantation to (42.49 ± 4.80) g/L (P＜0.01), alanine aminotransferase (ALT) decreased from (57.05±45.51) U/L to (44.86±29.19) U/L (P＜0.05),aspartate aminotransferase (AST) decreased from (39.14-±-15.07) U/L to (53.73 ± 24.98) U/L(P＞0.05).Congulation parameters were also improved, prothrombin time (PT) decreased from (16.15±3.01) s to (14.63±2.32) s (P＜0.01), fibrinofen (Fib) increased from (2.44±0.61) g/L to (3.00±0.81) g/L (P＜0.01).Compared with first transplantation group, the albumin level was higher in second autologous bone marrow stem cells transplantation group, which increased from (38.00±6.33) g/L to (42.49±4.80) g/L (P＜0.05), AST and ALT also improved obviously, and there was significant difference between two groups.Meanwhile, Child-Pugh scores decreased from (7.22±0.67) to (6.67±[0.71) (P＜0.05).But there was no significant difference in bilirubin, FIB and PT.ConclusionThe second transplantation of autologous bone marrow stem cells could further improve liver function and maintain symptoms remission of liver cirrhosis.

Objective To observe the inhibitory effect on metastasis and growth of pancreatic cancer in mice by injection of KAI1 gene in vivo. Methods Pancreatic cancer cell line MiaPaCa Ⅱ was used to construct the nude mice models bearing tumors, then the mice were divided into normal saline group, Ad group and Ad-KAI1 group. Since the 10th days of model construction, the Ad-KAI1 was injected every 7 d and repeated twice, then the tumor size, the weight of liver, lung and their pathologic changes were evaluated. Results The tumor sizes were not significantly different between the three groups. The weight of lung and liver of Ad-KAI1 group was (0.366±0.041) g and (1. 35±0.21) g, respectively; the weight of lung and liver of Ad group was (0.57±0.065) g and (1.58±1.828) g, respectively; the weight of lung and liver of control group was (0.66±0.13)g and (1.95±0.344)g, respectively. The difference between Ad-KAI1 group and control group was significantly different (t = 5.984, P < 0. 05), and there was no significant difference between Ad group and control group (t=1.089, P > 0.05). The number of pulmonary, liver and lymph node metastasis in Ad-KAI1 group was (1±1), (2±1) and (2±2), respectively; in Ad group was (6±2), (5 ±1), (10±2), respectively; in control group was (7±2), (6±2), (11±3), respectively. The difference between Ad-KAI1 group and control group was significantly different (t = 7.44, 4.34, 8. 16, P < 0.05), while the difference between Ad group and control group was not significantly different (t=0.92, 0.64, 0.42, P >0.05). Conclusions KAI1 gene directly injected into tumors of nude mice may inhibit the growth and metastasis of pancreatic cancer.

Objective To evaluate the contraception efficacy, mode of bleeding, side effects and other positive effects of drospirenone-ethinylestradiol (Yasmin) in healthy Chinese women. Methods This was a multicenter, randomized, control study of 768 healthy Chinese women who consulted about contraception. The subjects were randomized into Yasmin group (30 μg ethinylestradiol plus 3 mg drospirenone, 573 cases) or desogestrel group (30 μg ethinylestradiol plus 150 μg desogestrel, 195 cases) with the ratio of 3: 1. Each individual was treated for 13 cycles. Further visits were required at cycle 4, cycle 7, cycle 10 and cycle 13 of treatment. Weight, height, body mass index were evaluated at each visit. The menstrual distress questionnaire (MDQ) was given to the women at baseline, visit 3 (cycle 7) and visit 5 (after cycle 13). Results The values of basal features were similar between two groups (P> 0.05). The Pearl index (method failure) of Yasmin was 0. 208/hundred women year which was lower than that of desogestrel (0. 601/hundred women year). The mode of bleeding was similar between two groups after trial without showing any significant difference. According to MDQ subscale, the improvement of water retention and increasing appetite during inter-menstrual period and water retention and general well-being during menstrual period in the Yasmin group ( -0. 297, -0. 057, 0. 033, 0. 150 respectively) was more obvious than that in the desogestrel group ( - 0. 108, 0. 023, 0. 231, - 0. 023 respectively) with a significant difference (P < 0. 05 ). Some other values which improved in beth two groups, especially the improvement of breast tenderness and pain and skin abnormality in Yasmin group (18.0%, 89/494; 12. 6%, 62/494) was more distinct than that in desogestrel group (11.3%, 19/168; 5.4%, 9/168). The mean weight increased in desogestrel group (0. 57 kg) while it decreased in Yasmin group ( -0. 28 kg) with a significant difference (P < 0. 01 ). Conclusions Both Yasmin and desogestrel have good efficacy on contraception and similar modes of menstrual bleeding. Yasmin is better than desogestrel in terms of weight control and premenstrual syndrome of oral contraceptive.

To investigate the presence of integrated Epstein-Barr virus (EBV) DNA in the NK/T cell lymphoma (NKTCL) ge-nome and analyze the integration information in the genome of NKTCL cell lines. Methods: PCR and in situ hybridization were used to detect EBV infection in five EBV (+) NK/T samples and four EBV (-) NK/T samples provided by the biobanks of the First Affiliated Hospi-tal of Zhengzhou University. Whole-genome DNA of the samples was sequenced and subjected to bioinformatics analysis. Whole-ge-nome sequence alignment was used to identify the EBV integration sequence. BLAST analysis was used to compare EBV fasta files of the samples and EBV fasta library. CREST software was used to extract softclip reads, filter all paired reads, and enumerate their distri-bution on chromosomes. The integrated genomics viewer (IGV) was used to compare the distribution of reads in partial regions of chromosome. PCR was used to amplify the high-frequency integration region of the EBV DNA. The amplified fragments were sanger se-quenced. Results: EBV DNA and EBER expression were detected in five EBV (+) NK/T samples but not in the four EBV (-) NK/T samples. Sequencing depth, coverage depth, proportion of coverage, and proportion of alignment all met the requirements for subsequent re-search. Sequence alignment revealed that the captured sequences were viral sequences. Filtered reads were most numerous in EBV (+) NKTCL cell line SNK, YTS, and EBV (+) nasal NKTCL tissue. The reads were non-randomly enriched in chromosome 2. EBV DNA inte-gration in the 400 bp region of chr2:30234084-30234483 caused insertion or deletion in the chr2p23.1 site. Conclusions: EBV DNA is highly integrated in the chr2p23.1 site of EBV (+) NKTCL cells and may affect the expression of related genes.

Objective: To investigate the expression and clinical significance of programmed death-ligand 1 (PD-L1), programmed death-ligand 2 (PD-L2), and their receptor programmed cell death protein 1 (PD-1) in EBV-positive T/NK lymphoproliferative disease [Epstein-Barr virus-positive T/natural killer (NK)-cell lymphoproliferative disease, EBV(+)-T/NK-LPD]. Methods: The pathological paraffin-embedded tissues of 17 patients with EBV(+)-T/NK-LPD from the First Affiliated Hospital of Zhengzhou University from January 2013 to December 2017 were collected. These patients include 12 males and 5 females, aged 10-82 years old, the average age being 29 years, 4 people in gradeⅠ, 7 in gradeⅡ, 3 in gradeⅢ, and 3 people with hydroa vacciniforme-like lymphoproliferative disorders. Immunohistochemical SP method was used to detect the expression of PD-1, PD-L1, and PD-L2 in human EBV(+)-T/NK-LPD tissues. The relationship between PD-1, PD-L1, PD-L2 expression, and clinicopathological parameters, pathological grades and prognosis were analyzed by Fisher's exact probabilities and Spearman rank correlation. Result: After statistical analysis, the results showed that in 17 cases of tissue samples, there were 12 cases with positive PD-1 expression, 6 cases with positive PD-L1 expression and 5 cases with positive PD-L2 expression. There was no significant correlation between PD-1 and PD-L2 expression and prognosis (P>0.05). PD-L1 expression showed a positive correlation with prognosis (P<0.05). There was no significant correlation between the expression of PD-L1 and PD-L2 with age, sex, as well as LDH and Ki-67 levels (P>0.05). Moreover, there was no significant correlation of PD-1 and PD-L2 expression with pathological grade (r=0.141, r=-0.149, both P>0.05). However, there was a negative correlation between the PD-L1 expression and pathological grade (r=-0.563), and the correlation between the PD-L1 ex-pression and pathological grade was statistically significant (P<0.05). Conclusions: PD-1, PD-L1, and PD-L2 are abnormally expressed in the pathological tissues of EBV(+)-T/NK-LPD. Although there was no significant correlation between the expression of PD-1 and prognosis or pathological grade, it was significantly higher in EBV+T/NK-LPD. PD-1/PD-Ls associated signaling pathway is expected to be a potential new target for EBV(+)-T/NK-LPD immunotherapy.

Despite being a common therapy for hepatocellular carcinoma (HCC), insufficient thermal ablation can leave behind tumor residues that can cause recurrence. This is believed to augment M2 inflammatory macrophages that usually play a pro-tumorigenic role. To address this problem, we designed d-mannose-chelated iron oxide nanoparticles (man-IONPs) to polarize M2-like macrophages into the antitumor M1 phenotype. In vitro and in vivo experiments demonstrated that man-IONPs specifically targeted M2-like macrophages and accumulated in peri-ablation zones after macrophage infiltration was augmented under insufficient microwave ablation (MWA). The nanoparticles simultaneously induced polarization of pro-tumorigenic M2 macrophages into antitumor M1 phenotypes, enabling the transformation of the immunosuppressive microenvironment into an immunoactivating one. Post-MWA macrophage polarization exerted robust inhibitory effects on HCC progression in a well-established orthotopic liver cancer mouse model. Thus, combining thermal ablation with man-IONPs can salvage residual tumors after insufficient MWA. These results have strong potential for clinical translation.

OBJECTIVE To explore the pharmaceutical service model in multidisciplinary diagnosis and treatment （MDT） of rare diseases in children. METHODS Clinical pharmacists of West China Second University Hospital （hereinafter referred to as “our hospital”） participated in the process of MDT of children’s rare diseases. Clinical pharmacists took part in the entire diagnosis and treatment process of children and established the MDT pharmaceutical service model of children’s rare diseases by formulating drug treatment plans based on evidence-based practice， improving the accessibility of drugs， pharmaceutical monitoring and drug treatment management. RESULTS From January 2021 to April 2022， clinical pharmacists of our hospital had participated in a total of 39 cases of rare diseases MDT in children， including 21 hospitalized children with rare diseases and 18 outpatient com children with rare diseases， involving a total of 23 rare diseases. Clinical pharmacists completed 45 pharmaceutical zhanglingli@scu.edu.cn rounds and 26 pharmaceutical consultations for rare diseases inpatients， 25 outpatients’ MDT and 5 pharmaceutical outpatient service for outpatients with rare diseases， 38 medication educations for inpatients and outpatients with rare diseases and 25 follow-up services for out-of-hospital patients. There were 24 cases （61.54%） of off-label drug use， involving 13 rare diseases and 16 therapeutic drugs， among which off-label drug use registration of 11 drugs had been completed or was in progress. The temporary purchase evaluations of 3 drugs had been completed； 268 cases of medical insurance drug and high-value drug prescription had been reviewed. CONCLUSIONS Our hospital have primarily established a loop pharmaceutical service model of MDT for children with rare diseases， which covers inpatients and outpatients. The model improves the availability and standardization of clinical application of therapeutic drugs， and diagnosis and treatment level for children with rare diseases in our hospital.