Objective To develop a sensitive and reliable RP-HPLC method for the simultaneous determination of nine nucleosides including uracil, cytidine, guanine, uridine, adenine, guanosine, thymidine, adenosine and 2'-deoxyadenosine from Fritillaria taipaiensis P. Y. Li that had been cultivated in different producing areas; To compare the contents of these nucleosides from different producing areas. Methods The analysis was performed on a Venusil MP C18 (2) column (4.6 mm×250 mm, 5 μm) with a gradient of methanol-water at a flow rate of 1.0 mL/min;the detective wavelength was set at 260 nm; the column temperature was set at 35 ℃. Results Uracil, cytidine, guanine, uridine, adenine, guanosine, thymidine, adenosine and 2'-deoxyadenosine were obtained in the good linear range of 0.269 5–16.17 μg/mL, 0.132 1–7.927 5 μg/mL, 0.095 5–5.73 μg/mL, 1.16–69.6 μg/mL, 0.48–28.8 μg/mL, 0.571 5–57.15 μg/mL, 0.526–52.6 μg/mL, 3.307 5–198.45 μg/mL, 0.530 5–31.83 μg/mL, respectively (r≥0.999 5);the recovery was in the range of 96.49%–101.65%(RSD≤2.92%). Conclusion The contents of the nine nucleosides from different producing areas have differences. Fritillaria taipaiensis P. Y. Li from Xianyi Village, Chengkou County, Chongqing City, whether the cultivated ones or wild ones, contain the highest level of nucleosides. The established method can provide references for the perfection of quality standard for Fritillaria taipaiensis P. Y. Li.

Objective To establish a HPLC method for the determination of quercetin from different parts of L. cuncata(Dum. Cours.)G.Don.,including roots,old branches,young shoots,leaves and seeds,and to build the fingerprints. Methods The HPLC method determination of quercetin and fingerpints were establised. Results The restults showed that there were great differences be?tween the quercetin contents from different parts,with the highest contents found in seeds,followed by young shoots,leaves,old branches,and roots. The similarities of HPLC fingerprints of the medicinal material were quite different from the parts of L. cuncata (Dum.Cours.)G.Don.. The similarities were all above 0.95 for L. cuncata(Dum.Cours.)G.Don. samples,roots,old branches and young shoots below 0.85,leaves and seeds similarities below 0.60. Conclusion It was concluded that different parts(such as roots, old branches,young shoots,leaves,seeds)of L. cuncata(Dum.Cours.)G.Don. should be divided in clinical and productive practice so as to supply the scientific basis for enhancing the curative effect and reasonable utilization of the resource of L. cuncata(Dum.Cours.)G. Don.

ObjectiveTo investigate the effect of bone marrow-derived mesenchymal stem cells (MSCs) on gene and protein expression of toll-like receptor, TLR2/4 of the rat alveolus macrophages so as to discuss the effect of MSCs transplantation on the early inflammatory mediators and the regulatory mechanism of the inflammatory cascade.MethodsThe alveolus macrophages were randomly divided into the PBS control group (n = 24), MSCs control group (n = 24), lipopolysaccharide (LPS) control group (n=24) and LPS + MSCs treatment group (n =24).All groups were taken time-points of 1, 3,6, 12 hours.Positive cell rate of MSCs with surface mark and the expression level of TLR2/4 protein on macrophage were detected by flow cytometry.The mRNA level of TLR2/4 expression on macrophage was detected by RT-PCR and the concentrations of TNF-o in cell culture supernatants by ELISA at the different time points.ResultsCompared with the PBS control group and MSCs control group, TLR2/4 mRNA and protein levels were significantly up-regulated, markedly increased at one hour and peaked at 12 hours (P ＜0.01) and the TNF-α concentrations in cell culture supernatants were significantly up-regulated and peaked at 6 hours in the LPS control group (P ＜ 0.01).Compared with the LPS control group, mRNA and protein level of TLR2/4 and TNF-o concentrations in the cell culture supernatants of the LPS + MSCs group were markedly down-regulated (P ＜ 0.01).ConclusionsWith LPS stimulation, TLR2/4 mRNA and protein expressions as well as TNF-αt concentration increase, when the peak of the TNF-αt concentration appears slightly earlier, indicating that there may be a positive feedback loop between TLR2/4 expression and TNF-ot.MSCs can significantly reduce TLR2/4 mRNA and protein expressions as well as TNF-α concentration after LPS stimulation, indicating that MSCs can break up the positive feedback loop.

BACKGROUNDMost of the researches about lung cancer have focused on the polymorphisms of single gene rather than the cooperation of the genes. The aim of this study is to investigate the correlation of gene polymorphisms of cytochrome P450 2E1 (CYP2E1) Rsa I and cytochrome P450 1A1 (CYP1A1) independently and in combination with the risk of lung cancer.

METHODSA case-control study which included 91 lung cancer cases and 91 hospital-based controls was carried out, and all of them were the Han nationality in Guangzhou district. Blood samples were collected for detecting CYP2E1 Rsa I and CYP1A1 gene polymorphisms by polymerase chain reaction (PCR) and restriction fragment length polymorphisms (RFLP).

RESULTSNo significant difference was observed in genotype frequencies of CYP2E1 Rsa I and CYP1A1 Msp I between case and control groups. Odd ratio was 5.48 and 1.51 respectively without significant difference (P > 0.05). Individuals with CYP2E1 Rsa I C1C1 and CYP1A1 m2m2 genotype had 3.00-fold risk of developing lung cancer compared with those who had CYP2E1 Rsa I C1C2 and CYP1A1 m1m1 genotype (P < 0.05, 95% CI 1.03-8.78).

CONCLUSIONSSingle polymorphism of CYP2E1 and CYP1A1 doesn't increase the risk of developing lung cancer, but the combination of two genetic polymorphisms significantly increases the risk of developing lung cancer.

Objective: To investigate the abnormalities of the blood system in landfill workers. Methods: A cohort study was conducted for 224 landfill workers who were followed up for 6 consecutive years with abnormal routine blood test results and a low platelet count as the outcome events. The life-table method was used to analyze the incidence rates of these two outcome events, and the incidence rates were compared between first-and second-line workers. Results: A total of 71 workers had abnormal routine blood test results, among whom 29 had abnormal leukocyte count, 14 had abnormal erythrocyte count, 40 had abnormal platelet count, 17 had abnormal hemoglobin, and 29 had a reduction in platelet count. For these landfill workers, the 6-year abnormal rate of routine blood test results was 43.2%, and the incidence rate of low platelet count within 6 years was 13.5%. The first-line workers had a significantly lower abnormal rate of routine blood test results than the second-line workers (P=0.020) , and the relative risk of abnormal routine blood test results in the first-and second-line workers was 0.592. There was no significant difference in the incidence rate of low platelet count between the two groups of workers (P≥0.05) . Conclusion The landfill has an adverse effect on the blood system of landfill workers, and the second-line workers have greater impairment than the first-line workers.

Objective To investigate major risk factors of mixed hemorrhoid,and provide scientific basis for primary prevention of the disease.Methods A hospital-based 1:1 matched case-control study method was adopted.A total of 341 patients who were diagnosed with mixed hemorrhoid in the Department of Anorectal Diseases,the Affiliated Hospital of Chengdu University of Traditional Chinese Medicine from June 2015 to February 2016 was recruited as case group.Another 341 non-mixed-hemorrhoid patients with the same gender proportion and age difference within 5 years old were taken as control group.Self-designed questionnaires were applied to face-to-face interview these subjects.The questionnaires included occupational factors,living habits,defecation related factors,disease history,reproductive history,family history and basic knowledge of mixed hemorrhoid.SPSS 20.0 software was used for single-factor analysis.Results It turned out that the factors related to occurrence of hemorrhoid were as followed:education level(x2 =15.431),working position(x2 =18.078),duration of single working position(OR=3.345),alcohol intake(OR=3.269),smoking(OR=1.852),spicy food intake(OR =2.409),less physical exercise(OR =1.522),defecation posture(OR =1.750),defecation time(x2 =7.516),defecation frequency(x2 =8.405),stool shape(x2 =8.004),obesity(OR=1.618),reproductive history(OR=2.211) and hemorrhoid family history of first-degree relatives(OR=1.763);The correlation intension (OR value) between anal fissure,anal pruritus and mixed hemorrhoid was 0.564 and 2.714 respectively;While tea drinking,perianal abscess or anal fistula were in no relation to mixed hemorrhoid onset.Conclusion The onset of mixed hemorrhoid is resulted from joint efforts of occupational factors,living habits,defecation habits and reproductive history,etc.Anal fissure,history of anal pruritus and preanal eczema are closely related to mixed hemorrhoid,and genetic factors may get involved as well.

Objective:To analyze the clinical effect of hyaluronidase injection through the facial artery in the treatment of vascular embolism such as skin ulceration and necrosis after cosmetic injection.Methods:Hyaluronidase was injected through facial artery in 13 patients who were diagnosed with vascular embolism after facial injection from January 2019 to May 2020. The facial artery was punctured with 22-gauge arterial blood collection needle or 19/23-gauge disposable venous infusion needle. The angle between the needle body and the skin varies depending on the patients’ weight, ranged 30°-45°. The needle was advanced slowly and pushed forward by 2-3 mm when blood backflow appeared in the needle core. After confirming the successful puncture of the facial artery, 0.5-1.5 ml hyaluronidase was slowly injected into the facial artery. The time of skin relaxation, tenderness relief, ulcer healing and wound recovery were observed. The pigmentation was observed and the Vancouver Scar Scale (VSS) was used to score the scars after 3-12 months.Results:A toal of 13 patients with vascular complications of hyaluronidase filler were retrospectively reviewed. The patients were 18-45-year-old(mean age, 35 years) and received hyaluronidase filler at private clinics. There were 12 women and 1 man. The time from onset to visit was 14 h to 4 d, with an average time of 2.5 d. Hyaluronidase was most commonly injected into the nasolabial folds (54%, 7 of 13). The second-ranked area is the nasalroot (23%, 3 of 13). These patients had skin swelling, necrosis, ecchymosis or black scabs during or after hyaluronidase injection. Some patients showed skin lesions combined with oral ulcer. After percutaneous facial arterial hyaluronidase injection, the local skin tissue injuries of the 13 patients were improved in time. The time of skin relaxation was (0.77±0.25) d, the time of tenderness relief was (1.23±0.64) d, the time of ulcer healing was (3.14±0.64) d and the time of wound recovery was (5.85±0.86) d. Patients were followed up for 3-12 months, with an average of 7 months. One patient had slight scar (VSS score of 1), two patients had only mild pigmentation (VSS score of 0), and the other ten patients had no scar and pigmentation (VSS score of 0).Conclusions:It is effective to improve local microcirculation and reduce skin tissue injury after percutaneous facial artery hyaluronidase injection in the treatment of skin injury caused by facial filler injection.

Objective:To analyze the clinical effect of hyaluronidase injection through the facial artery in the treatment of vascular embolism such as skin ulceration and necrosis after cosmetic injection.Methods:Hyaluronidase was injected through facial artery in 13 patients who were diagnosed with vascular embolism after facial injection from January 2019 to May 2020. The facial artery was punctured with 22-gauge arterial blood collection needle or 19/23-gauge disposable venous infusion needle. The angle between the needle body and the skin varies depending on the patients’ weight, ranged 30°-45°. The needle was advanced slowly and pushed forward by 2-3 mm when blood backflow appeared in the needle core. After confirming the successful puncture of the facial artery, 0.5-1.5 ml hyaluronidase was slowly injected into the facial artery. The time of skin relaxation, tenderness relief, ulcer healing and wound recovery were observed. The pigmentation was observed and the Vancouver Scar Scale (VSS) was used to score the scars after 3-12 months.Results:A toal of 13 patients with vascular complications of hyaluronidase filler were retrospectively reviewed. The patients were 18-45-year-old(mean age, 35 years) and received hyaluronidase filler at private clinics. There were 12 women and 1 man. The time from onset to visit was 14 h to 4 d, with an average time of 2.5 d. Hyaluronidase was most commonly injected into the nasolabial folds (54%, 7 of 13). The second-ranked area is the nasalroot (23%, 3 of 13). These patients had skin swelling, necrosis, ecchymosis or black scabs during or after hyaluronidase injection. Some patients showed skin lesions combined with oral ulcer. After percutaneous facial arterial hyaluronidase injection, the local skin tissue injuries of the 13 patients were improved in time. The time of skin relaxation was (0.77±0.25) d, the time of tenderness relief was (1.23±0.64) d, the time of ulcer healing was (3.14±0.64) d and the time of wound recovery was (5.85±0.86) d. Patients were followed up for 3-12 months, with an average of 7 months. One patient had slight scar (VSS score of 1), two patients had only mild pigmentation (VSS score of 0), and the other ten patients had no scar and pigmentation (VSS score of 0).Conclusions:It is effective to improve local microcirculation and reduce skin tissue injury after percutaneous facial artery hyaluronidase injection in the treatment of skin injury caused by facial filler injection.

OBJECTIVE：To establish a meth od for the simultaneous determination of 7 active components in Mori Australis Cortex and Mori Cortex from different sources in Chongqing area ，so as to provide reference for improving the quality control standards of Mori Australis Cortex and Mori Cortex and comparing the equivalence of their quality. METHODS ：HPLC method was used to determine the contents of neochlorogenic acid ，mulberroside A ，chlorogenic acid ，astragalin，kaempferol，morusin and isoquercetin in 58 batches of Mori Australis Cortex and Mori Cortex. The chromatographic column was Diamonsil C 18 with mobile phase consisted of 0.1％ formic acid solution-acetonitrile （gradient elution ） at the flow rate of 1.0 mL/min. The detection wavelength was 280 nm，column temperature was 30 ℃，and the injection volume was 10 μL. Using SPSS 22.0 software， independent sample t-test，principal component analysis and cluster analysis were used to analyze the content difference of the above-mentioned 7 active components in Mori Australis Cortex and Mori Cortex. RESULTS ：There was a good linear relationship between the peak area and the concentration of the above 7 active components （r≥0.999 0）. The RSDs of precision ，stability（24 h），repeatability，durability and recovery were less than 3％. The average contents of neochlorogenic acid ，mulberroside A ， chlorogenic acid ， astragalin， kaempferol， morusin and 023-58576130。E-mail：1025473978@qq.com isoquercetin in Mori Australis Cortex were 0.304，22.462， 1.730，1.308，1.593，2.842 and 0.657 mg/g，respectively. Those of Mori Cortex were 0.305，22.995，2.486，2.438， 2.916，4.158 and 1.264 mg/g，respectively. The results of independent sample t-test showed that only the content of kaempferol in the above 7 active components of Mori Australis Cortex and Mori Cortex had significant difference （P＜0.05）. The results of principal component analysis and cluster analysis showed that there was no significant difference in the contents of above 7 active components between Mori Australis Cortex and Mori Cortex. CONCLUSIONS：The established HPLC method is simple ，sensitive and accurate ，which can provide a reference for improving the quality control standard of Mori Australis Cortex and Mori Cortex. Mori Australis Cortex and Mori Cortex have certain quality equivalence in main active components ，and the Mori Australis Cortex from M. australis and M. cathayana can be used as a substitute for the Mori Cortex.

Objective: To explore the clinical application of the deep inferior epigastric artery perforator flap in bilateral breast reconstruction of patients with breast cancer. Methods: Deep inferior epigastric artery perforator flap was applied for breast reconstruction in four cases of breast cancer patients received radical or modified radical surgery in Hunan Provincial Cancer Hospital. All patients are female, age ranged from 31 to 53 years old (36.2±5.9). Results: The length of flap was (15.1±0.4) cm, the width of flap was (12.6±0.3) cm, the thickness of flap was (4.3±0.5) cm. The length of pedicle was (12.6±0.3) cm, the outer diameter of artery was (1.8±0.2) mm, the outer diameter of vein was (2.1±0.4) mm.The average weight of flaps was 235 g(ranged from 195 g to 335 g). In one case flap fat necrosis occurred and in other one donor site fat necrosis was noted. The two flaps both healed with dressing treatment and no other complications were found. The reconstructed breasts′shape, texture and elasticity were good and no flap contracture deformation happened. Only linear scar left in the donor sites, the function of abdomen did not affected. All 4 patients were followed up for 14 to 33 months (21.7 months on average) with satisfied result. No local recurrence happened. Conclusions Free deep inferior epigastric artery perforator flap is suitable to reconstruct bilateral breast for breast cancer patients.