AIM:To prepare a compound as the chemical reference substance of Guangjinqiancao Zonghuangtong Capsule.METHODS:To apply general column chromatography combined with preparative HPLC to isolate the target compound,to use analytic HPLC to determine the purity,stability and its content in the capsule,and to employ spectroscopic analysis (UV,IR,ESI-MS,1H-NMR,13C-NMR,DEPT,1H-13CCOSY,1H-1HCOSY,1DHOHAHA.1D.NOE,HMBC) to elucidate the structure of the isolated compound.RESULTS:The obtained compound was identified as isoschaftoside with the purity of over 99%, which was stable within 3 months at ambient temperature.As for isosehaftoside solution.it was stable within 8 h at ambient temperature.Its content in the capsule was above 3.0%.CONCLUSION:Isoschaftoside is a qualified reference substance for analytic assay ofGuangjinqiancao Zonghuangtong Capsule,and can be isolated from Desmodium styracifolium(Osb.)Merr.

To reduce the risk of 3′-terminal mismatch between primers and template and increase the sensitivity of polymerase chain reaction (PCR) in the detection of variable region of DNA. Methods: A pair of special primer(WU,WD) was designed to amplify a fragment of HBV DNA P gene by PCR. Other 2 similar pairs of primer (MU1, MD1, MU2, MD2) were obtained by knocking off 1 or 2 bases at the 3′-terminal of WU and WD. (1) Special primers (WU, WD) and degeneracy primers(WU, WD, MU1, MU2, MD1, MD2) were used to amplify 27 samples respectively by PCR under the same condition. The sensitivity of each PCR was compared. (2) Using degeneracy primers, serum HBV DNA was amplified from 4 patients who were resistant to lamivudine. The PCR products were sequenced to evaluate the effect of the 3′-terminal mismatch of primers upon PCR. Results: (1) The sensitivity of special primers and degeneracy primers were 70.4%(19/27) and 85.2%(23/27) respectively (P<0.05). (2) The sequencing analysis of the PCR products suggested that the 3′-terminal mismatch of primers caused false negative in the PCR detection. Conclusion: When amplifying the variable region of DNA, the false negative result can be avoided by using 3′-terminus shifted degeneracy primers.

Objective Xiangqingzhitong spray ( XQ spray) is a kind of class V new Chinese drugs .This paper studied the anti-inflammatory and analgesic effects of XQ spray to provide experiment evidence for clinical application . Methods Mice were divided into blank control A group , yunnan baiyao aerosol A group , voltaren A group and low , middle and high dose A group of XQ spray.Analgesic effects were studied by observing effect of XQ spray for pain threshold in electrical stimulation test .Mice were divided into blank control B group , yunnan baiyao aerosol B group , voltaren B group and low , middle and high dose B group of XQ spray .Analgesic effects were studied by observing the effects of XQ spray for pain threshold in tail pain tenderness test .Mice were divided into blank control C group , yunnan baiyao aerosol C group , voltaren C group and low,middle and high dose C group of XQ spray .Anti-inflammatory effects were observed by the models of ear swelling induced by xylene in mice.Rats were divided into blank control D group , yunnan baiyao aerosol D group , voltaren D group and low, middle and high dose D group of XQ spray .Anti-inflammatory effects were observed by the models of paw edema induced by carrageenin in rats . Results Electrical stimulation test showed that middle and high dose A group of XQ spray could enhance the pain threshold and the analgesia percentage reached 90%after the administration of 15 min and 30 min.Compared with control group , significant difference was found in medium and high dose A group (P<0.05).High dose B group of XQ spray could enhance pain threshold in tail pain tenderness test markedly after the administration of 15 min and 30 min and the difference was significant compared with control group ( P<0.01).Middle dose C group could inhibit the ear swelling induced by xylene in mice and the difference was significant compared with control group ([4.41 ±4.55]mg vs [9.15 ±3.94]mg,P<0.05).High dose C group could inhibit the ear swelling induced by xylene in mice and significant difference was found compared with control group ([3.62 ±3.56]mg vs [9.15 ±3.94] mg,P<0.01). High dose D group could inhibit the paw edema induced by carrageenin in rats after the administration of 30 min and there was signifi-cant difference compared with control group ([1.19 ±0.94]mg vs [2.54 ±1.28]mg,P<0.01). Conclusion XQ spray has sig-nificant anti-inflammatory and analgesic effects .

Objective: To reduce the risk of 3'-terminal mismatch between primers and template and increase the sensitivity of polymerase chain reaction (PCR) in the detection of variable region of DNA. Methods: A pair of special primer(WU, WD) was designed to amplify a fragment of HBV DNA P gene by PCR, Other 2 similar pairs of primer (MU1, MD1, MU2, MD2) were obtained by knocking off 1 or 2 bases at the 3'-terminal of WU and WD. (1) Special primers (WU, WD) and degeneracy primers(WU, WD, MU1, MU2. MD1, MD2) were used to amplify 27 samples respectively by PCR under the same condition. The sensitivity of each PCR was compared. (2) Using degeneracy primers, serum HBV DNA was amplified from 4 patients who were resistant to lamivudine. The PCR products were sequenced to evaluate the effect of the 3'-terminal mismatch of primers upon PCR. Results: (1) The sensitivity of special primers and degeneracy primers were 70. 4%(19/27) and 85. 2% (23/27) respectively (P

Objective:To investigate the influencing factors and to build a model for predicting successful labor induction with cervical ripening balloon (CRB) in primipara.Methods:312 cases of primipara in late trimester of pregnancy induced by CRB between January 2018 and April 2019 in the First Affiliated Hospital of Shenzhen University were reviewed retrospectively. Factors including gravidity, age, body height, body weight, body mass index (BMI), the cervical Bishop score, neonatal birth weight, pregnant complication and other factors were collected to analyze the effect on successful labor induction.Results:Pregnancy times, cervical Bishop score and height in primipara were positively correlated with the success rate of induced labor, while BMI and neonatal weight were negatively correlated with the success rate of induced labor. The area under receiver operating characteristic (ROC) curve was 0.849 and diagnostic threshold was 0.725, with a sensitivity of 80.7% and a specificity of 75.7%.Conclusions:Among the primipara, the gravidity and cervical Bishop score are the most significant factors influencing the successful labor induction. These factors can be used as the predictive indexes for the outcome of labor induction along with pregnancy times, height, BMI and neonatal birth weight, which is beneficial to reduce the artificial interventions and improve the success rate of labor induction.

OBJECTIVE:To study the effects of flow components C6 and C7 in n-butyl alcohol extract from the leaves of Ces-trum Nocturnum(CN)on the proliferation and apoptosis of human gastric cancer cell SGC7901. METHODS:C6 and C7 were ob-tained by using different ratio of chloroform and methanol(1:9,1:7)to the gradient elution of CN leaves. After cultured with 0 (blank control),5,10,20,40,80 μg/ml C6 and C7 for 72 h,inhibitory effect of C6 and C7 on the proliferation of SGC7901 was determined by MTT assay. Inhibitory rate and IC50 were calculated. After SGC7901 were cultured with 10 μg/ml C6 and C7 for 72 h,colony formation assay was utilized to detect the effects of C6 and C7 on the cell colony formation,and the rate of colony for-mation was calculated. In addition,Wright/Giemsa and Hoechst33258/PI staining assay were used to observe the change of cytomor-phology. RESULTS:MTT showed that C6 and C7 had inhibitory effect on the proliferation of SGC7901 to different extent;inhibi-tory rates were 22.1%-80.0% and 19.6%-79.7%,and IC50 were 16.4,18.05 μg/ml,respectively. Compared with blank control group,colony formation rate of C6 and C7 group decreased,with statistical significance(P<0.05). The number of apoptotic cells was more in treatment group than in other groups. CONCLUSIONS:Flow components C6 and C7 in n-butyl alcohol extract from the leaves of CN can inhibit the proliferation of SGC7901 cells and induce the apoptosis of them.

Objective To observe the effect of recombinant human growth hormone on flap survival and angiogenesis through preoperative local subcutaneous continuous injection of rhGH in narrow pedicle flap.Methods 40 rats were divided into two groups at random,named as groups Ⅰ and Ⅱ,20 rats each.A narrow pedicle flap was designed on the left back of rat in groups Ⅰ and Ⅱ.Then group Ⅰ was subcutaneously injected with rhGH [0.4 IU/(kg · d)] for 3 days.Another group did not have any injection.After 3 days,we established a model of narrow pedicle flaps in the left back of animals.After operation,the flaps of groups Ⅰ and Ⅱ were injected with rhGH [0.4 IU/(kg · d)]for 7 days.The survival percentage of the flap was calculated at 7'th day.Specimens were collected at the diffent point of time.Expression of VEGF,CD34 was detected by immunohistochemistry and ELISA.The MVD of the flaps was also calculated.Results According to the 7 days' observation after the surgery,the percentage of flap survival area for groups Ⅰ and Ⅱ was (100.00±0.00)％,and (94.00±1.10)％ (P＜0.05).The expression of VEGF,CD34 and the MVD values of group Ⅰ was higher than that of group Ⅱ at the same point of time (P＜0.05),especially on the day of surgery.Conclusions Preoperative and local subcutaneaous continuous injection of rhGH on flaps can enhance the expression of VEGF and CD34 and microvascular formation,mitigate the damage of ischemia and hypoxia to flap during the postoperative period,and therefore improve the survival of narrow pedicle flap.

Objective To study the proliferation,collagen production and related gene expression in keloids and normal skin fibroblast.Methods Isolated primary cells of keloid fibroblasts (KFb,n=12) and normal human dermal fibroblasts (NFb,n=12) were identified,the cell viability and proliferating potential and the cell cycle were detected,and the difference on the collagen synthesis between KFb and NFb were compared.The expression of cell cycle-associated genes such as p21,p16,and p27 was dectected by real-time fluorescent quantitative PCR.Results The phase contrast optical microscopy imaging showed that both KFb isolated from keloid tissues and NFb from normal skin tissues possessed classic and similar fibroblast morphology.But there was a significant difference between cell proliferation,Hyp [(2.30±0.10) μg/ml vs.(1.66±0.13) μg/ml,P＜0.05] and collagen levels [(17.19±0.75) μg/ml vs.(12.37±0.94) μg/ml,P＜0.05].Compared with NFb,KFb exhibited more percentage of G2/M phase cells [(5.90±0.62)％ vs.(16.94 ％±1.93)％,P＜0.05]and less percentage of G0/G1 phase cells [(90.24 ±2.27)％ vs.(75.65±1.92)％,P＜0.05].Cell cycle related genes p16,p21 and p27 were low expressed.Collagen type Ⅰ was highly expressed at mRNA levels in KFb than that in NFb [0.84±0.11,1.32±0.2,1.69±0.12,4.33±0.27 in KFb vs.1.43±0.13,2.56±0.26,2.89±0.37,1.40±0.12 in NFb,P＜0.05].Conclusions There are cell dysfunction and abnormal cellular dynamics in keloid fibroblasts.The formation of keloid likely involves aberrant interactions of some genes that affected its development at different extents.

Objective To analyze the changes of HIV‐1 viral load and virological efficacy of treatment effects for AIDS patients treated with antiretroviral therapy from 2009 to 2014 in Lincang City .Methods Monitored the HIV‐1 viral load for 13 491 cases of AIDS patients treated with highly active antiretroviral therapy(HAART) from 2009-2014 in Lincang City of Yunnan and analyze the monitoring data .If the patients had treated with HAART were still with viral load greater than 1 000 copy/mL ,the treatment was defined as a failed treatment or a virological failure .Results The total rate of virological failure was 14 .34% (1 935/13 491) . The rate of virological failure of children group was 15 .53% (50/322) ,and of adults group was 14 .31% (1 885/13 169) .There was no statistically significant difference between childeren and adults(χ2 =0 .38 ,P>0 .38) .The rate of virological failure in males was 16 .34% (1 156/7 076) ,and 12 .14% (779/6 415) in females ,the difference was statistically significant between men and female (χ2 =48 .16 ,P<0 .01) .Conclusion Antiretroviral treatment can delay the disease progression and improve the life quality .

Objective To explore clinical effect of repairing soft tissue defect in forearm, hand and foot with free super-thin anterolateral thigh perforator flaps. Methods At first the site of perforator vessels were determined by Doppler, then the flaps were designed and harvested with the site as center; the fascia lata and subcutaneous fat were removed by sandhill-likely only the 4.0 cm × 3.0 cm - 3.0 cm×2.5 cm disc-like fascia lata and dermis layer were reserved. 15 traumatic soft tissue defects including forearm, hand and foot were repaired with the ree super-thin antemlateral thigh perforator flaps. Results No vascular crisis happened and all skin grafts survived in donor sites. 2.0 cm×1.2 cm of the distal of flap was necrosis in 1 case and it was healed by dress changing. 15 cases were followed up 3 months-2 years and the average is 6 months. The contour and texture of all flaps were good and two point discrimination (2-PD) was about 8-10 mm of. Conclusions The contour and texture of free super-thin anterolateral thigh perforator flap are good, the feeling of recipient site recovered well, it's less injury for donor site and there is no reshaping for flap. It is a fineness donor site for repairing soft tissue defects in hand and foot.