Objective To establish a new RP-HPLC method for simultaneous determination of chlorogenic acid,forsythin, and baicalin in shuanghuanglian powder for injection after ultrasonic atomization. Methods Hypersil ODS2 C18(250 mmí4. 6 mm,5 μm) was used as the chromatographic column. The mobile phase was methanol-0. 2% phosphate acid solution (4060). Flow rate was 1. 0 mL·min-1 . Sample volume was 5μL. Column temperature was 30℃. Detection wavelength was 324 nm at 0-10 min and 277 nm at 10-25 min. Results Contents of chlorogenic acid,forsythin, and baicalin had good linear relationship with the respective peak area (r≥0. 999 7) within the scope of the sample volume. The RSD was <2% for precision, reproducibility, and stability. Recovery rate was 98. 50%-101. 12% (n=6). Conclusion The method is rapid, accurate and reproducible, with high resolution. It can determine the content of three kinds of components at the same time. The three components in shuanghuanglian powder for injection did not change significantly before and after ultrasonic atomization.

OBJECTIVETo explore the expansion method of high purity NK cells from human peripheral blood and explore the changes in biological functions of NK cells after ex vivo expansion.

METHODSNK cells were isolated from peripheral blood mononuclear cells (PBMNCs) by using miniMACS (Magnetic cell-selection) and NK Cell Isolation Kit II, and cultured in SCEM (Stemline Hematopoietic Stem Cell Expansion Medium, Sigma) supplemented with 10% human AB serum and different combinations of interleukin (IL)-2 and/or IL-12, IL-15 for 15 days. Cultures were semi-exchanged with fresh media and cytokines every 3 days. Evaluation for cell expansion, phenotype, perforin and granzyme B mRNA expressions, and IFN-gamma secretion before and after the culture period.

RESULTSCD3(-) CD56(+) cells concentration increased from (11.2 +/- 5.2)% to (94.2 +/- 3.5)%. In group IL-2 + IL-15 and IL-2 + IL-15 + IL-12 group, cells were expanded 50.5 +/- 4.3 and 52.3 +/- 6.7 - fold, respectively, being significantly higher than that in other three groups [(15.4 +/- 1.1 fold in IL-2 group, 19.9 +/- 3.9 fold in IL-2 + IL-12 group, 6.1 +/- 1.0 fold in control group)] (P<0.01), but no significant difference between each other (P>0.05). The purity of CD3(-) CD56(+) NK cells was over 94% in all groups except the control. The perforin and granzyme B mRNA expressions of expanded NK cells in four experimental groups were significantly higher than those of before expansion (P<0.01) and the expressions in IL-2 + IL-15 and in IL-2 + IL-12 + IL-15 group were significant higher than in other three groups (P<0.01) while no significant difference between each other (P>0.05). IFN-gamma levels in the supernatants of four experiment groups were significantly higher than that in control group (P<0.01) and its levels order was IL-2 + IL-15 + IL-12 group > IL-2 + IL-12 group > IL-2 + IL-15 group > IL-2 group (P<0.01).

CONCLUSIONHigh purity NK cells isolated by negative selection using miniMACS can be efficiently expanded with IL-2 + IL-15, and their biological functions were enhanced.

Cell Culture Techniques ; Cell Proliferation ; Cell Separation ; Cells, Cultured ; Granzymes ; metabolism ; Humans ; Interferon-gamma ; metabolism ; Interleukin-12 ; pharmacology ; Interleukin-15 ; pharmacology ; Interleukin-2 ; pharmacology ; Interleukins ; pharmacology ; Killer Cells, Natural ; cytology ; drug effects ; immunology ; metabolism ; Perforin ; metabolism

China ; HIV Infections ; prevention & control ; transmission ; Health Knowledge, Attitudes, Practice ; Health Personnel ; statistics & numerical data ; Humans ; Infectious Disease Transmission, Patient-to-Professional ; prevention & control ; Occupational Exposure ; prevention & control

OBJECTIVETo establish a rapid quantitative analysis method for the content of chlorogenic acid and solid content in the extraction liquid concentration process during the production of Reduning injection by using the near-infrared (NIR) spectroscopy, in order to reflect the concentration state in a real-time manner and really realize the quality control of concentrating process of the extraction and concentration process.

METHODThe samples during the Jinqing extraction liquid concentration process were collected. After the removal of abnormal samples, the spectra pretreatment and the wave band selection, the quantitative calibration model between NIR spectra and chlorogenic acid HPLC analytical value and solid content was established by using PLS algorithm, and unknown samples were predicted.

RESULTThe correlation coefficients between the chlorogenic acid content and the solid content were respectively 0.992 1 and 0.994 0, and the correlation coefficients of the verification model were respectively 0.994 4 and 0.998 4, with the root mean square error of calibration (RMSEC) of 0.814 6 and 2.656 1 and the root mean square error of prediction (RMSEP) of 0.704 6 and 1.876 7 respectively, and the relative standard errors of predictions (RSEP) were 6.01% and 2.93% respectively.

CONCLUSIONThe method is simple, rapid, nondestructive, accurate and reliable, thus could be adopted for the fast monitoring of the chlorogenic acid content and the solid content during the concentration process of Reduning injection extraction liquid.

Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Quality Control ; Spectroscopy, Near-Infrared ; methods

AIMTo determine the entrapment efficiency of breviscapine nanoliposomes.

METHODSRetrodialysis method was used to determine the entrapment efficiency of breviscapine nanoliposomes. The drug recovery and the sample recovery were considered to verify the feasibility of the method.

RESULTSThe drug recovery was (99.6 +/- 0.6)%. The sample recovery was (99.9 +/- 0.8)%. The entrapment efficiency of breviscapine nanoliposomes was (75.3 +/- 2.3)%.

CONCLUSIONThe method used in this study is simple, applicable and accurate for determination of the entrapment efficiency of breviscapine nanoliposomes.

Asteraceae ; chemistry ; Drug Carriers ; Flavonoids ; administration & dosage ; chemistry ; isolation & purification ; Liposomes ; Nanotechnology ; Particle Size ; Plants, Medicinal ; chemistry ; Technology, Pharmaceutical ; methods

AIM: To investigate the clinical characteristics of patients with systemic lupus erythematosus ( SLE) complicated with retinopathy. METHODS: Totally 121 cases of SLE patients treated in our hospital from February 2015 to February 2017 were selected,including 30 cases with retinopathy(observation group) and 91 cases without retinopathy(control group), the clinical manifestations and laboratory examination results of the two groups were compared. RESULTS: ln the observation group, there were 6 patients with bilateral retinopathy and 24 patients with monocular retinopathy. Cotton retinal exudation, retinal vascular occlusion and retinal hemorrhage lesions were common, accounted for 33%, 25% and 19%. The incidence of skin rash, skin vasculitis and Raynaud's phenomenon in the observation group were 63%, 47% and 37%, respectively, which were significantly higher than those in the control group (P<0.05). There were no significant difference in the incidence of mucosal ulcer, arthritis, neuropsychiatric symptoms and pleurisy between the observation group and the control group(P>0.05). The positive rate of anti ds-DNA in the observation group was 63%, which was significantly higher than that in the control group (P<0.05). The observation group and the control group urine protein > 3 +, anti Sm antibody positive and rRNP positive and antiphospholipid antibody percentage differences were not statistically significant(P>0.05). The SLEDAI score of the observation group was 20.14 (9, 30) points, which was significantly higher than that of the control group (P<0. 05). The modified BenEzra score of the observation group was 10 04士3.15,and was positively correlated with the SLEDAI score (rs=0.706,P<0.05). CONCLUSION: SLE with fundus retinopathy patients mainly manifest as cotton wool spots, retinal vascular occlusion and retinal hemorrhage; rashes, vasculitis, Raynaud's phenomenon,and anti ds-DNA antibodies are common in SLE patients with retinopathy. The modified BenEzra score plays an important role in the evaluation of SLE disease activity and retinal vascular lesions.

This study was purposed to explore the changes in biological functions of human peripheral blood derived NK Cells after ex vivo expansion with different combinations of interleukin IL2 and/or IL12, IL15. According to different combination of cytokines, cultured NK cells were divided into 4 groups: group IL2, group IL2 + IL12, group IL2 + IL15 and group IL2 + IL15 + IL12. The group in which NK cells were cultured without cytokines was used as control. The cytotoxicity of cultured NK cells to target K562 cells was determined by using cell counting kit-8; the level of IFN-gamma in supernatants of NK cell culture was detected by ELISA; the perforin and granzyme B mRNA expressions were assayed by competitive quantitative RT-PCR. The results showed that the cytotoxicity of expanded NK cells in groups cultured with cytokines at different E:T ratio was significantly higher than that in group without cytokines (p < 0.01), although the cytotoxicity of NK cells in IL2 + IL15 + IL12 group seem to be slightly higher than that in IL2 + IL15 group, but there was no statistic difference (p > 0.05). The IFN-gamma levels in the supernatants of NK cell culture in the presence of cytokines significantly increased, and the IFN-gamma levels in IL2 + IL15 + IL12 group and IL2 + IL12 group were significantly higher than that in others (p < 0.01). The expressions of perforin and granzyme B mRNA of expanded NK cells in groups cultured with cytokines was significantly higher than that in control group (p < 0.01), and was consistent with cytotoxicity of NK cells. It is concluded that there are differences in the functions of NK cells cultured with different cytokines. IL2 and IL15 have synergistic effect on strengthening cytotoxicity of NK cells and promoting cell expansion. However, the main function of IL12 promotes NK cells to secrete IFN-gamma, which plays a role in immunoregulation.
Humans ; Interferon-gamma ; secretion ; Interleukin-12 ; administration & dosage ; pharmacology ; Interleukin-15 ; administration & dosage ; pharmacology ; Interleukin-2 ; administration & dosage ; pharmacology ; K562 Cells ; Killer Cells, Natural ; drug effects ; immunology

Primary central nervous system germ cell tumors (CNS-GCTs) in children and adolescents have unique clinical features and methods of treatment compared with those in adults. There is little information about Chinese children and adolescents with CNS-GCTs. Therefore, in this study we retrospectively analyzed the clinical features and treatment outcome of Chinese children and adolescents with primary CNS-GCTs. Between January 2002 and December 2012, 57 untreated patients from a single institution were enrolled. They were diagnosed with CNS-GCTs after pathologic or clinical assessment. Of the 57 patients, 41 were males and 16 were females, with a median age of 12.8 years (range, 2.7 to 18.0 years) at diagnosis; 43 (75.4%) had non-germinomatous germ cell tumors (NGGCTs) and 14 (24.6%) had germinomas; 44 (77.2%) had localized disease and 13 (22.8%) had extensive lesions. Fifty-three patients completed the prescribed treatment, of which 18 underwent monotherapy of surgery, radiotherapy, or chemotherapy, and 35 underwent multimodality therapies that included radiotherapy combined with chemotherapy or surgery combined with chemotherapy and/or radiotherapy. PEB (cisplatin, etoposide, and bleomycin) protocol was the major chemotherapy regimen. The median follow-up time was 32.3 months (range, 1.2 to 139 months). Fourteen patients died of relapse or disease progression. The 3-year event-free survival (EFS) and overall survival rates for all patients were 72.2% and 73.8%, respectively. The 3-year EFS was 92.9% for germinomas and 64.8% for NGGCTs (P = 0.064). The 3-year EFS rates for patients with NGGCTs who underwent monotherapy and multimodality therapies were 50.6% and 73.5%, respectively (P = 0.042). Our results indicate that multimodality therapies including chemotherapy plus radiotherapy were better treatment option for children and adolescents with CNS-GCTs.
Adolescent ; Antineoplastic Agents ; therapeutic use ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bleomycin ; administration & dosage ; Central Nervous System Neoplasms ; therapy ; Child ; Child, Preschool ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; statistics & numerical data ; Disease-Free Survival ; Etoposide ; administration & dosage ; Female ; Humans ; Male ; Neoplasm Recurrence, Local ; Neoplasms, Germ Cell and Embryonal ; therapy ; Retrospective Studies ; Survival Rate ; Treatment Outcome

BACKGROUNDTo avoid the irritation of tendons and soft tissues as well as hardware-related problems, we designed an intramedullary fixation with bioabsorbable rods for the treatment of the metacarpal shaft fractures.

METHODSFive patients with nine shaft fractures of the fourth and fifth metacarpi were treated with intramedullary absorbable implants and followed up with an average of 4.2 months postoperatively.

RESULTSAt final follow-up, all patients achieved fracture union with no signs of inflammatory or subcutaneous effusion. There was no shortening, angulatory, or rotatory deformity. There was almost full active extension range of motion (ROM) of the metacarpophalangeal joints while the active flexion ROM of these joints was 80.7 ± 9.6°. Compared with the contralateral hand, the grip strength of the injured hand was 94.0 ± 9.6%. X-rays showed that the arch of the second to fifth metacarpal heads was smooth. There were no intramedullary lytic changes and soft tissue swellings.

CONCLUSIONThe intramedullary absorbable implants are a safe, simple, and practical treatment for fourth and fifth metacarpal fractures with good early clinical outcomes and no significant complications.

Absorbable Implants ; Adolescent ; Adult ; Fracture Fixation, Intramedullary ; methods ; Fractures, Bone ; surgery ; Humans ; Internal Fixators ; Male ; Metacarpal Bones ; injuries ; surgery ; Range of Motion, Articular ; physiology ; Treatment Outcome ; Young Adult

Objective: To analyze the relationship between single nucleotide polymorphisms (SNP) of Notch signaling pathway and susceptibility to lung cancer. Methods: The present study was a hospital-based case-control study. All 1 121 patients of lung cancer diagnosed by histopathology three hospitals in Fujian and Nanjing were selected as cases from January 2006 to December 2012. At the same time, 1 121 healthy population from other departments of the hospital to visit patients or community, excluding those with tumor, chronic disease, and immediate family members of lung cancer, were enrolled in control group. A uniform questionnaire was used to collect general information. Matrix-assisted laster desorption ionization time of flight mass spectrometry (MALDI-TOF-MS) was used to identify the polymorphisms of 9 SNP (Notch3 rs3815188, Notch4 rs915894, Notch4 rs520692, DLL1 rs1033583, JAG1 rs8708, JAG2 rs9972231, HEY1 rs1046472, HEY2 rs3734637, HES2 rs11364) in 1 121 lung cancer patients and 1 121 healthy controls. The association between SNP and lung cancer was analyzed by χ² and logistic regression model. Results: The average age of cases and controls was (58.70±10.73) and (58.98±10.85) years old. The OR for genotype AC carriers of HEY1 rs1046472 was 0.80 (95%CI: 0.66-0.97) when comparing with genotype CC. The OR for genotype AC+AA carriers of HEY1 rs1046472 was 0.81 (95% CI: 0.67-0.98) when comparing with genotype CC. The OR for genotype AC carriers of HEY2 rs3734637 was 0.82 (95%CI: 0.67-0.99) when comparing with genotype AA. In the stratified analysis, Notch3 rs3815188, DLL1 rs1033583, JAG1 rs8708, JAG2 rs9972231, HEY1 rs1046472, HEY2 rs3734637, HES2 rs11364 were associatied with the risk of lung cancer, P were 0.041, 0.030, 0.043, 0.003, 0.004, 0.026 and 0.038, respectively.The interactions analysis done by logistic regression model showed JAG1 rs8708 and family history, JAG2 rs9972231 and BMI had interaction in the study, OR were 2.07 (95% CI:1.21-3.52) and 1.73 (95% CI:1.21-2.47), respectively. Conclusion Notch3 rs3815188, DLL1 rs1033583, JAG1 rs8708, JAG2 rs9972231, HEY1 rs1046472, HEY2 rs3734637 and HES2 rs11364 were significantly associated with susceptibility to lung cancer.