Analysis of chemical constituents and components absorbed into plasma of Ardisia crenata based on UPLC-QE-HF-MS/MS / 中国药房

OBJECTIVE To analyze the chemical constituents and components absorbed into plasma of the extract of Ardisia crenata and to elucidate its possible pharmacodynamic material basis. METHODS Overall， 12 rats were randomly assigned to the blank group （n＝6） and A. crenata group （n＝6） by the paired comparison method. The drug was administered once daily in the morning and afternoon for three days. Serum samples were prepared from serum after redosing on 4th day. The UPLC-QE-HF-MS/ MS was used to analyze and identify the chemical constituents in A. crenata extract and serum samples. Compound Discoverer 3.0 was employed for retention time correction， peak identification， and peak extraction. According to the secondary mass spectrometry information， the Thermo mzCloud online and Thermo mzVault local databases， referring to the relevant literature and control quality spectrum information were used to preliminarily identify the chemical constituents and components absorbed into plasma of A. crenata. RESULTS A total of 34 compounds were identified from the extract of A. crenata， mainly coumarins， flavonoids， organic acids， amino acids， including bergenin， quercetin， gallic acid， L-pyroglutamic acid， etc. Besides， 5 components absorbed into plasma were identified from serum samples: L-pyroglutamic acid， syringic acid， bergenin， cinnabar root saponin A， and mycophenolic acid. CONCLUSIONS L-pyroglutamic acid， syringic acid， bergenin， cinnabar root saponin A， and mycophenolic acid may act as the pharmacodynamic material basis of A. crenata.

Analysis of Chemical Constituents as Flavonoids and Coumarins in Radix Ardisiae from Different Sources / 中国药房

OBJECTIVE：To identif y and analyze the flavonoids and coumarins in Radix Ardisiae from different sources. METHODS：UPLC-QE-HF-MS/MS was adopted. The determination was performed on Zorbax Eclipse-C 18 column with mobile phase consisted of acetonitrile- 0.1％ formic acid solution （gradient elution ）at the flow rate of 0.3 mL/min. The column temperature was 30 ℃，and the temperature of injector was 4 ℃. The sample size was 2 µL；ESI source was applied in negative and positive scanning ion mode ，the heater temperature was 325 ℃，the sheath gas pressure was 45 arb，the auxiliary gas pressure was 15 arb，the purge gas pressure was 1 arb，the electrospray voltage was 3.5 kV，the capillary temperature was 330 ℃， S-lens RF level was 55％，scan mode was first-order full sca m/z 100-1 500，data-dependent secondary mass spectrometry scanning （dd-MS2，Top N ＝10），the resolution was 70 000 （first mass spectrometry ） ， 17 500 （secondary mass spectrometry），the collision mode was high-energy collision dissociation. Through retrieving foreign and domestic databases as ChemSpider ，mzCloud，mzVault，PubChem，the structure of the compound was identified on the basis of related literatures and reference data ，and the conten ts were compared. RESULTS & CONCLUSIONS：A total of 47 components were separated from Radix Ardisiae of 3 kinds of sources as Ardisia crenata Sims，A. crispa（Thunb.）A. DC. ，A. crenata Sims var . bicolor （Walk）C. Y. Wu et C. Chen. A total of 17 flavonoids were identified ，including 9 flavonols （quercetin 3-O-rhamnoside-7-O-glucoside， myricetin， rutin， mauritanin， kaempferol， quercetin， isorhamnetin， quercetin，mearnsitrin），3 flavan-3-ols [（-）-epigallocatechin，catechin，epigallocatechin gallate ）2 dihydroflavonoids [fustin ， eriodictyol] and 3 other types [ 3-（2，3-dihydro-benzo[1，4]dioxin-6-yl）-7-hydroxy-2-trifluoromethyl-chromen-4-one，methadone， oriciacridone F] ，10 coumarins {bergenin ，（[ 7-hydroxy-4-methyl-2-oxo-2H-chromen-6-yl）oxy]acetic acid ，[7-（carboxymethoxy）- 4-methyl-2-oxo-2hydroxychromo-3-yl]acetic acid ，4，9-dihydroxy-7H-furo[3，2-g]chromen-7-one，6，7-dihydroxy-4-methylcoumarin， esculetin，fraxetin，7，8-dihydroxy-4-methylcoumarin，4-methylumbelliferyl glucuronide ，scoparone}. Results of content analysis showed that in flavonoids and coumarins ，there were 5 common components in Radix Ardisiae from 3 kinds of sources ，i.e. bergenin（peak 2），[7-（carboxymethoxy）-4-methyl-2-oxo-2-hydroxychromo-3-yl] acetic acid （peak 5），methadone（peak 16）， quercetin（peak 18），oriciacridone F （peak 26）；the contents of common components were significantly different. In addition to 5 common components ，there were 22 different chemical components ，which were compounds corresponding to peaks 1，3，4， 6-15，17，19-25 and 27，respectively. Among them ，compounds corresponding to peaks 3，6，8 and 23 were only found in A. crenata Sims var. bicolor（Walk）C. Y. Wu et C. Chen ；compounds corresponding to peaks 12-15，19 were only found in A. crispa （Thunb.）A. DC. UPLC-QE-HF-MS/MS method can efficiently ，accurately and quickly identify the flavonoids and coumarins in Radix Ardisiae from different sources.

Oncologic outcomes of early stage cervical cancer performed operation by different laparoscopic surgical procedures: analysis of clinical data from mutiple centers / 中华妇产科杂志

Objective:To evaluate the oncologic outcomes of different laparoscopic radical hysterectomy.Methods:From January 2011 to December 2014, the laparoscopic operation cases of cervical cancer at stage Ⅰb1, Ⅰb2, Ⅱa1 and Ⅱa2, including the histologic subtypes of squamous-cell carcinoma, adenocarcinoma and adenosquamous carcinoma, were collected in five clinical centers. The data were divided into two groups according to the surgical procedures, that is, modified laparoscopic-vaginal radical hysterectomy (mLVRH) and total laparoscopic radical hysterectomy (TLRH). The overall survival rate (OS), disease-free survival rate (DFS) at 5 years were retrospectively analyzed in this study.Results:There were 674 cases in total, including 377 cases of mLVRH, 297 cases of TLRH. (1) The OS at 5 years: the mLVRH was 96.1% and the TLRH was 92.0%, and the mLVRH was higher than that of TLRH （ P=0.010）. Stratify analysis， including stage of disease （Ⅰb1 and Ⅱa1）， histologic subtypes （squamous-cell carcinoma, adenocarcinoma）， lymph node metastasis， revealed that, ① Stage of disease: in stage Ⅰb1, the OS at five years of mLVRH was higher than that in TLRH group （98.6% vs 93.6%, P=0.012）. In stage Ⅱa1, there was significant difference between the two groups, the OS at five years of mLVRH and TLRH were 93.6% and 77.6% （ P=0.007）. ② Histologic subtypes: for the OS at five years of squamous-cell carcinoma, mLVRH and TLRH were 96.1% and 92.3%, and there was significant difference （ P=0.046）； for adenocarcinoma, the OS at five years were 91.0% and 88.6%， and there was no difference between two groups （ P=0.230）. ③ Lymph node metastasis： the mLVRH and TLRH with lymph node metastasis, the OS at five years were 98.6% and 96.4%; the mLVRH and TLRH without lymph node metastasis, the OS at five years were 89.3% and 80.8%. There were no significant differences between the two groups,respectively ( P=0.156， P=0.093）. (2) The DFS at 5 years: there was no significant difference between mLVRH and TLRH (94.1% vs 90.9%, P=0.220). Stratify analysis for stage of disease, the mLVRH group was higher than that in the TLRH group in stage Ⅰb1 (97.0% vs 92.8%, P=0.039). However, for stage Ⅱa1, there was no significant difference between mLVRH and TLRH group （88.2% vs 75.8%, P=0.074）. Conclusions:The results of this retrospective study indicated that different laparoscopy surgical procedures had diverse oncologic outcomes. The OS at 5 years of the mLVRH is superior to the TLRH. The DFS at 5 years in Ⅰb1 stage, the mLVRH is higher than the TLRH. Therefore, the modified laparoscopy is still an alternative surgery for early cervical cancer patients when following the principle of no-tumor-exposure.

Comparison of the therapeutic effect of TGF-β1 gene therapy and traditional nonsurgical treatment on the rat model of postpartum stress urinary incontinence / 中国实验动物学报

Objective To study the effect of TGF?β1 gene therapy on the rat model of postpartum stress urinary incontinence and explore a novel non?operative treatment of this disease. Methods Two hundred and forty 6?month old SD female rats were used to prepare the model of postpartum stress urinary incontinence by vaginal dilation with a water sac. 148 rats from the 185 successfully prepared model rats were selected, and randomly divided into 5 groups: the TGF?β1 gene therapy, clentuterol treatment, electric stimulation therapy, injection of empty vector plasmid, and non?treated groups. In addition, 20 normal rats were selected as blank control group. Sneeze test and urodynamic test were conducted, the pelvic floor pubococcygeus muscle contractile force/muscle weight ratio was calculated, serum TGF?1 was detected by ELISA, and TGF?1 protein was detected by immunohistochemistry at 1, 21, 42 and 63 days after the treatment. Results At 21 days after treatment, all the maximum bladder capacity, leak point pressure, and urine or contractile force / muscle weight ratio of the TGF?β1 gene therapy group showed even better effects than those of the electrical stimulation group, but the differences were statistically not significant ( P>0?05 ) . Conclusions TGF?β1 gene therapy shows good therapeutic effect on the rat models of postpartum stress urinary incontinence, suggesting that TGF?β1 gene therapy may become a new type of non?surgical treatment for this disease.

Effect of IGF-1 gene and electric stimulation therapy on the rat model of postpartum stress urinary incontinence / 中国实验动物学报

Objective To study the effect of IGF-1 gene therapy and electric stimulation therapy on the rat models of postpartum stress urinary incontinence, and explore the ideal treatment for this disease.Methods 240 SD female rats were used to establish the model of postpartum stress urinary incontinence by water sac vaginal dilation.148 model rats were randomly selected from 185 successful models and divided into 5 groups:IGF-1 gene therapy, clenbuterol treatment, electric stimulation therapy, injection of empty vector plasmid, and untreated groups.Besides, 20 non-modeled rats were used as blank control group.Urodynamic test was performed, pelvic floor pubococcygeus muscle/muscle weight ratio was calculated, and serum biochemical indices (LDH, CK) were detected, and the morphological changes of pubococcygeus muscle fibers were observed by light microscopy at 1, 21, 42 and 63 days after treatment.Results At 21 days after treat-ment, the maximum bladder capacity, leak point pressure, the contractile force/muscle weight ratio in the IGF-1 group and electric stimulation treatment group were significantly better (P>0.05), and the differences between the IGF-1 group and electric stimulation group were not significant ( P>0.05 ) .Conclusions The effect of IGF-1 gene therapy and electric stimulation on the rat models of postpartum stress urinary incontinence is better than that in the drug therapy group and oth-er groups.

Study on the effects of gonadotropin-releasing hormone analogues in the inhibition of ovarian cancer transplanted tumors and in the protection of ovarian function after chemotherapy on nude mice / 中华妇产科杂志

Objective To investigate the influence of gonadotropin-releasing hormone (GnRH) analogues on ovarian cancer and ovarian function in vivo.Methods ES-2 cells were cultured and xenotransplanted into 36 nude mice,which were divided into 6 groups:normal saline (NS) group:NS 0.1 nd/day subcutaneous injection,and then NS 0.2 ml/week peritoneal injection; cisplatin (DDP) group:NS 0.1 ml/day subcutaneous injection,and then DDP 5 mg/kg ( diluted to 0.2 ml ) per week peritoneal injection; goserelin group:100 μg goserelin ( diluted to 0.1 ml) per day subcutaneous injection,and then NS 0.2 ml/week peritoneal injection; goserelin + DDP group:100 μg goserelin ( diluted to 0.1 ml) per day subcutaneous injection,and DDP 5 mg/kg (diluted to 0.2 ml) per week peritoneal injection; cetrorelix group:100 μg cetrorelix (diluted to 0.1 ml) per day subcutaneous injection and NS 0.2 ml/week peritoneal injection; cetrorelix + DDP group:100 μg cetrorelix (diluted to 0.1 ml) per day subcutaneous injection and DDP 5 mg/kg ( diluted to 0.2 ml) per week peritoneal injection.All the peritoneal injection started from subcutaneous injection one week later.To compare the weight of nude mice,the volumes of transplanted tumors,the expression of Ki-67 antigen in transplanted tumors,the estrus,the ratio of atretic follicles,the ratio of primary and preantral follicles,the levels of serum anti-Mullerian hormone ( AMH ),folliclestimulating hormone ( FSH),estradio ( E2 ) and progesterone (P) in each group.Results There were no significant difference in the weight of nude mice among 6 groups ( P ＞ 0.05 ),which on day 29 in NS group was ( 19.8 ±2.2) g,DDP group (20.5 ± 1.4) g,gosereline group ( 19.6 ±0.9) g,goserelin + DDP group ( 19.7 ± 1.6) g,cetrorelix group (20.7 ±2.2) g,and cetrorelix + DDP group ( 19.0 ± 1.7) g.The tumor volumes of different groups on the 12th day:NS group (241 ± 179) mm3,DDP group (78 ±20) mm3,gosereline group (78 t±55) mm3,goserelin + DDP group (64 ±48) mm3,cetrorelix group (78 ±64) mm3,or cetrorelix + DDP group (70 ± 19) mm3,in which there were significant difference between NS group and the other groups ( P ＜ 0.05 ) ; and the same result was obtained on day 15,19,22,26 and 29 ( P ＜ 0.05 ).The expression of Ki-67 in NS group was ( 33 ± 10 ) ％,in which it was higher than those in DDP group 3.5％,goserelin group 8.8％,goserelin + DDP group 1.5％,cetrorelix group (23 ± 11 ) ％,or cetrorelix + DDP group ( 8 ± 6 ) ％ ( P ＜ 0.05 ).The ratio of primary and preantral follicles in goserehn group was (71.5 ± 8.1 ) ％,in goserelin + DDP group was (62.4 ± 4.1 ) ％,in cetrorelix group was (71.2 ± 7.4) ％,and in cetrorelix + DDP group was (63.8 ±3.1 )％,in which they were much higher than that in DDP group ( 47.0 ± 4.8 ) ％ ( P ＜ 0.05 ).The level of AMH in goserelin group was ( 98 ± 27 ) ng/ml,which was much higher than that in NS group (66.2 ± 17.4) ng/ml (P ＜0.05),while there were no difference in the levelsof FSH,E2 or P among different groups ( P ＞ 0.05).Conclusion GnRH analogues could inhibit the growth of transplanted tumors in nude mice,meanwhile increase the secretion of AMH,decrease the frequencies and prolong the lasting time of estrus,decrease the ratio of atretic follicles,raise the ratio of primary and preantral follicles,which may be protect the ovarian function of nude mice.

Comparison outcomes of three surgical procedures in treatment of severe pelvic organ prolapse and analysis of risk factors for genital prolapse recurrence / 中华妇产科杂志

Objective To investigate clinical significance and application of modified pelvic floor reconstruction developed by Peking Union Medical College Hospital ( MPFR ) in treatment of severe pelvic organ prolapse (POP) by comparing the effectiveness, quality of postoperative sexual life, life satisfaction and risk factors for POP recurrence with the following two surgical procedures: traditional total vaginal hysterectomy with anterior-posterior colporrhaphy (TVH-APC) and total vaginal hysterectomy with lateral colporrhaphy and sacrospinous ligament fixation and vaginal bridge repair and episiotomy (TVH-LC-SSLFVBR-EP). Methods Totally 173 patients with severe POP and at least two compartments defects of pelvic floor underwent surgeries in the study, 86 patients (group A) were treated by MPFR with polypropylene mesh application, 58 (group B) were treated by TVH-APC, and 29 patients (group C) were treated by TVH-LC-SSLF-VBR-EP. Peri-operative data and outcomes of postoperative courses at 6, 12, 18 months were collected and analyzed, in the meantime, the risk factors of recurrence were studied. Results (1) No statistical difference was observed among the above 3 groups in terms of length of operation, amount of blood loss, length of hospital stay, and morbidity after surgery ( P ＞ 0.05). ( 2 ) Cost hospitalization was ( 11 448 ±3049) Yuan in group A, which was significantly higher than (7262 ± 1607) Yuan in group B and (7140 ± 1817 ) Yuan in group C (P ＜ 0.05 ). (3) The length of vaginal cuff of (7.5 ± 1.4) cm in group A and ( 5.6 ± 1.1 ) cm in group C were significantly longer than (7.1±0.6) cm in group B ( P＜0.05). The width of vaginal cuff of (4.3±0.3) cm in group A was larger than (3.4±0.3) cm in group B and (3.3±0.4) cm in group C (P＜0.05). (4) The recurrence rate at 12 months after surgery was 12.8% (11/86)in group A, which was similar with 17.2%(5/29) in group C (P＞0.05) and significantly less than 36.2% (21/58) in group B (P＜0.05). The rate of active sexual life of 16.3% (14/86) in group A was significantly higher than 1.7% (1/58) in group B and 0 in group C ( P ＜ 0. 05 ). The index of life quality improvement at 12 months after surgery was 48±12 in group A, which was no less than 53±16 in group C ( P＞0.05) and higher than 27 ± 9 in group B ( P＜0.05). (5 ) Mesh rejection was observed in 6 patients in group A within 3 months after surgery, while the posterior vaginal wall was exclusively involved. No difference was found in urinary retention or urinary incontinence among three groups (P ＞0. 05 ). (6) The severe degree of POP, type of surgical procedure ( TVT-APC), anterior compartment defect of pelvic floor,and early days of performing pelvic floor reconstruction surgeries were high risk factors for POP recurrence (P＜ 0.05). Conclusions MPFR has a better curative effect and lower recurrence rate on patients with POP. It can help patients regain integrity of anatomical structure and functions of pelvic floor. TVH-LCSSLF-VBR-EP is also effective.