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OBJECTIVE: To develop an ultra-high performance liquid chromatography coupled with quadrupole tandem time-of-flight mass spectrometry method for simultaneous determination of nine trace D-amino acids in thymalfasin, combined with deuterated acid hydrolysis and precolumn-derivatization. METHODS: The sample was hydrolyzed by deuterated acid, followed by precolumn-derivatization with Nα-(2, 4-dinitro-5-fluorophenyl)-L-alaninamide (FDAA). The analysis was then performed on an ACCQ-TAGTM ULTRA C18 column (2.1 mm×100 mm, 1.7 μm), with mobile phase comprising water containing 0.1% formic acid-acetonitrile gradiently eluted at a flow rate of 0.3 mL•min-1. Nine D-amino acids in thymalfasin were correctly quantified using standard curves by Xevo G2-S Q-TOF coupled with electrospray ion source in positive ion mode. RESULTS: The standard curves of the nine D-amino acids derivatives had good linearity in the ranges of the tested concentrations (r>0.991 2). The limits of quantitation of all D-amino acids derivatives were as low as 0.05-0.30 pmol. The precision and recoveries met the requirements of Chinese Pharmacopoeia (2015). The contents of D-amino acids in the samples from five companies were determined to be 25.16-1 322.01 μg•g-1. Among them, D-glutamate, D-aspartate, D-lysine and D-serine had higher contents, which were 1 105.45-1 322.01, 614.15-740.50, 358.06-388.76 and 138.78-291.60 μg•g-1, respectively. CONCLUSION: The method is sensitive, efficient and reliable, working well for simultaneous determination of nine trace D-amino acids in thymalfasin, which provides a reference for the comprehensive control of racemic peptide impurities in this synthetic polypeptide drug.
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We utilized a multi-step derivatization gas chromatography-mass spectrometry method for the determination of common amino acid enantiomers, combined with deuterated hydrochloric acid hydrolysis, to identify nine trace D-amino acids in thymalfasin. We optimized the conditions for multi-step derivatization, the volume of reagent for redissolving samples, and the conditions for chromatography and mass spectrometry with isopropanol and trifluoroacetic anhydride as derivatization reagents, and validated the procedure, including sensitivity, linear range, precision, accuracy and recovery. Sixteen pairs of D/L-amino acids and glycine derivatives were separated within 29 min, with the limit of quantification as low as 0.09-2.79 μmol·L-1. Nine amino acid derivatives of thymalfasin showed a good linear relationship within the concentration range examined (r2>0.992 3). The precision results showed that RSD values were less than 10.90%. Accuracy test results of a reference substance ranged from 76.69% to 128.18%. Average recoveries of spiked samples ranged from 70.41% to 125.39%. For the nine D-amino acids assayed, D-Asp and D-Glu content in six batches of thymalfasin were highest, ranging 0.41%-0.49% and 0.25%-0.33%, respectively, with others less than 0.25%. The method is sensitive, efficient and reliable, available for seventeen common amino acids and their enantiomers, and works well with simultaneous determination of nine trace D-amino acids in thymalfasin, providing a reference for the comprehensive control of racemic peptide impurities in this synthetic polypeptide drug.
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Objective@#To investigate the effect of enhanced recovery after surgery program with thymalfasin in patients undergoing laparoscopic radical nephrectomy in the perioperative period to enhance the quality of rehabilitation and reduce the risk of surgical complications.@*Methods@#Retrospective analysis of the clinical data of 156 patients undergoing radical laparoscopic radical nephrectomy in the People's Hospital of Renshou County from August 2013 to June 2018 was conducted.According to the perioperative management plan, 74 patients were divided into control group(74cases) and observation group(82cases). Laparoscopic surgery was performed in both two groups.The perioperative period of the observation group was accelerated surgical rehabilitation, and thymalfasin was given before and after surgery.The control group used a traditional perioperative management program.The blood loss, postoperative recovery time and complications were compared between the two groups.@*Results@#The loss of hemoglobin during operation[(11.90±5.61)g/L]in the accelerated rehabilitation group was slightly lower than that of the control group[(17.47±5.31)g/L], and the indwelling time of plasma drainage tube, anal exhaust time and hospitalization time in the accelerated rehabilitation group were (35.96±4.47)h, (12.61±4.05)h, (121.45±4.30)h, respectively, which were significantly shorter than those in the control group[(46.81±4.55)h, (15.34±4.45)h, (193.69±5.48)h], the differences were statistically significant (t=6.351, 15.011, 4.012, 92.043, all P<0.05).@*Conclusion@#Enhanced recovery after surgery program with thymalfasin in patients undergoing laparoscopic radical nephrectomy is a safe and effective treatment.The use of thymalfasin method can accelerate the recovery of patients' immune function, and the accelerated rehabilitation program with thymalfasin can effectively reduce the probability of infection during surgery, which can effectively reduce the possibility of infection during operation, accelerate the recovery of physiological function, shorten the hospital stay of patients and improve the quality of rehabilitation of patients.
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Objective@#To explore the clinical efficacy of thymalfasin on the immune function and inflammatory response in the treatment of patients with severe pneumonia.@*Methods@#From June 2015 to June 2018, 86 patients with severe pneumonia in the Hospital of Zhejiang Provincial General Team of Armed Police were enrolled in the study.According to different treatment methods, they were divided into control group and treatment group, with 43 cases in each group.The control group was treated with cefoperazone/sulbactam sodium on the basis of conventional treatment.The treatment group was treated with thymalfasin on the basis of the treatment of the control group.The expression of monocyte human leukocyte antigen-DR (mHLA-DR), tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels were measured before and after treatment.At the same time, CD4+, CD8+, CD4+/CD8+ before and after treatment were examined in the two groups.The efficacy and bacterial clearance rate of the two groups were evaluated.The adverse reactions were recorded.@*Results@#After treatment, the levels of IL-6 and TNF-α in the two groups were significantly lower than those before treatment [treatment group: (44.9±11.8)ng/L, (42.9±13.1)ng/L vs. (86.5±27.9)ng/L, (79.6±28.6)ng/L, control group: (71.5±14.2)ng/L, (65.9±22.6)ng/L vs. (87.1±28.6)ng/L, (78.8±29.1)ng/L, t=9.005, 7.650, 3.203, 2.295, all P<0.05]. The expression level of mHLA-DR in the treatment group after treatment [(44.8±5.7)%] was significantly higher than that before treatment [(27.1±3.4)%, t=17.487, P=0.000]. The changes of the indicators in the treatment group were significantly better than those in the control group (t=9.447, 5.773, 8.725, all P<0.05). After treatment, the CD4+ level of the two groups were higher than those before treatment, and the CD8+ levels of the two groups were significantly lower than those before treatment, the differences were statistically significant(t=3.050, 3.429, 6.965, 13.327, all P<0.05), and the CD4+/CD8+ of the two groups were significantly increased (t=0.370, 3.314, all P<0.05). The indicators of the treatment group were improved more significantly than the control group (t=4.416, 12.355, 3.089, all P<0.05). The bacterial clearance rate of the treatment group was 88.89% (32/36), which was significantly higher than that of the control group [67.65% (23/34), χ2=4.686, P=0.030]. The clinical efficacy of the treatment group was 93.02%(40/43), which was significantly higher than 76.74%(33/43) of the control group (χ2=6.095, P=0.047). The incidence of adverse reactions between the two groups had no statistically significant difference (χ2=0.212, P=0.645).@*Conclusion@#Thymalfasin in the treatment of patients with severe pneumonia can improve the immune function, reduce the inflammatory response, improve the bacterial clearance rate and clinical efficacy, and will not increase the adverse reactions, so it is worthy of promoting.
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Objective To investigate the effect of enhanced recovery after surgery program with thymalfasin in patients undergoing laparoscopic radical nephrectomy in the perioperative period to enhance the quality of rehabilitation and reduce the risk of surgical complications.Methods Retrospective analysis of the clinical data of 156 patients undergoing radical laparoscopic radical nephrectomy in the People＇s Hospital of Renshou County from August 2013 to June 2018 was conducted.According to the perioperative management plan ,74 patients were divided into control group (74cases) and observation group(82cases).Laparoscopic surgery was performed in both two groups.The perioperative period of the observation group was accelerated surgical rehabilitation ,and thymalfasin was given before and after surgery.The control group used a traditional perioperative management program.The blood loss,postoperative recovery time and complications were compared between the two groups.Results The loss of hemoglobin during operation [(11.90 ±5.61)g／L]in the accelerated rehabilitation group was slightly lower than that of the control group [(17.47 ± 5.31) g／L], and the indwelling time of plasma drainage tube , anal exhaust time and hospitalization time in the accelerated rehabilitation group were (35.96 ±4.47) h,(12.61 ±4.05) h,(121.45 ±4.30) h,respectively,which were significantly shorter than those in the control group [(46.81 ±4.55)h,(15.34 ±4.45)h,(193.69 ±5.48)h], the differences were statistically significant ( t ＝6.351, 15.011, 4.012, 92.043, all P ＜0.05 ).Conclusion Enhanced recovery after surgery program with thymalfasin in patients undergoing laparoscopic radical nephrectomy is a safe and effective treatment.The use of thymalfasin method can accelerate the recovery of patients ＇immune function, and the accelerated rehabilitation program with thymalfasin can effectively reduce the probability of infection during surgery,which can effectively reduce the possibility of infection during operation ,accelerate the recovery of physiological function,shorten the hospital stay of patients and improve the quality of rehabilitation of patients .
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Objective To explore the clinical efficacy of thymalfasin on the immune function and inflammatory response in the treatment of patients with severe pneumonia.Methods From June 2015 to June 2018,86 patients with severe pneumonia in the Hospital of Zhejiang Provincial General Team of Armed Police were enrolled in the study.According to different treatment methods,they were divided into control group and treatment group,with 43 cases in each group.The control group was treated with cefoperazone/sulbactam sodium on the basis of conventional treatment.The treatment group was treated with thymalfasin on the basis of the treatment of the control group.The expression of monocyte human leukocyte antigen-DR (mHLA-DR),tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) levels were measured before and after treatment.At the same time,CD4+,CD8+,CD4+ /CD8 + before and after treatment were examined in the two groups.The efficacy and bacterial clearance rate of the two groups were evaluated.The adverse reactions were recorded.Results After treatment,the levels of IL-6 and TNF-α in the two groups were significantly lower than those before treatment [treatment group:(44.9 ± 11.8)ng/L,(42.9 ± 13.1) ng/L vs.(86.5 ± 27.9) ng/L,(79.6 ± 28.6) ng/L,control group:(71.5 ± 14.2)ng/L,(65.9 ±22.6)ng/L vs.(87.1 ±28.6)ng/L,(78.8 ±29.1)ng/L,t =9.005,7.650,3.203,2.295,all P ＜ 0.05].The expression level of mHLA-DR in the treatment group after treatment [(44.8 ± 5.7) ％] was significantly higher than that before treatment [(27.1 ± 3.4) ％,t =17.487,P =0.000].The changes of the indicators in the treatment group were significantly better than those in the control group (t =9.447,5.773,8.725,all P ＜ 0.05).After treatment,the CD4+ level of the two groups were higher than those before treatment,and the CD8+ levels of the two groups were significantly lower than those before treatment,the differences were statistically significant(t =3.050,3.429,6.965,13.327,all P ＜ 0.05),and the CD4 +/CD8 + of the two groups were significantly increased (t =0.370,3.314,all P ＜0.05).The indicators of the treatment group were improved more significantly than the control group (t =4.416,12.355,3.089,all P ＜ 0.05).The bacterial clearance rate of the treatment group was 88.89％ (32/36),which was significantly higher than that of the control group [67.65％ (23/34),x2 =4.686,P =0.030].The clinical efficacy of the treatment group was 93.02％ (40/43),which was significantly higher than 76.74％ (33/43) of the control group (x2 =6.095,P =0.047).The incidence of adverse reactions between the two groups had no statistically significant difference (x2 =0.212,P =0.645).Conclusion Thymalfasin in the treatment of patients with severe pneumonia can improve the immune function,reduce the inflammatory response,improve the bacterial clearance rate and clinical efficacy,and will not increase the adverse reactions,so it is worthy of promoting.
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Objective To investigate the clinical effect of radiofrequency ablation (RFA) combined with non-specific sequential immunotherapy (IM) for early hepatocellular carcinoma (HCC),and analyze the factors affecting prognosis of patients after RFA.Methods The prosepctive study was conducted.The clinicopathological data of 72 early HCC patients who were admitted to the People's Hospital of Guangxi Zhuang Autonomous Region from January 2009 to October 2015 were collected.Patients were divided into 3 groups by random number table:patients in group A underwent single RFA therapy;patients in group B underwent RFA + non-specific sequential IM (1-3 times);patients in group C underwent RFA + non-specific sequential IM (≥ 4 times).RFA was performed by the same doctors team,and non-specific sequential IM planning included thymalfasin + interleukin-2 (IL-2).Observation indicators:(1) treatment situations;(2) follow-up and survival;(3) analysis of prognostic factors after RFA.Follow-up using outpatient examination was performed to detect tumor recurrence and overall survival up to December 2015.Measurement data with normal distribution were represented as (x) ± s,and comparison among groups were evaluated with the ANOVA.Comparison of count data were analyzed using the chi-square test.The curve,rate and time of tumor recurrence after treatment,overall survival curve and time were respectively drawn and calculated by the Kaplan-Meier method,and the Log-rank test was used for survival analysis.The univariate analysis and multivariate analysis were respectively done using the COX proportional hazard regression model.Results Seventy-two patients were screened for eligibility,including 31 in group A,22 in group B and 19 in group C.(1) Treatment situations:patients in 3 groups underwent RFA,and contrast enhanced ultrasound showed complete tumors ablation at 5 days postoperatively.Patients in group B and C didn't have significant adverse reactions after RFA during IM therapy.(2) Follow-up and survival:72 patients were followed up for 2-66 months after treatment,with a median time of 34 months.The 1-year tumor recurrence rates after treatment in group A,B and C were respectively 19.4％,13.6％ and 10.5％,with no statistically significant difference (x2=0.714,P＞0.05).The median tumor recurrence times in group A,B and C were respectively 24.0 months,30.0 months and 33.0 months,with no statistically significant difference (x2 =3.283,P＞0.05).The median overall survival times in group A,B and C were respectively 46.0 months,56.0 months and 57.0 months,with a statistically significant difference (x2=7.079,P＜0.05).There were statistically significant differences between group A and group B and C (x2 =4.566,4.243,P＜0.05),and no statistically significant difference between group B and group C (x2 =0.078,P＞0.05).(3) Analysis of prognostic factors after RFA:results of univariate analysis showed that initial tumor,tumor number,Barcelona clinic liver cancer (BCLC)staging and sequential IM after RFA were related factors affecting prognosis of early HCC patients [hazard ratio (HR)=2.636,2.530,0.145,0.582,95％ confidence interval (CI):1.218-5.703,1.110-5.767,0.041-0.517,0.321-0.867,P＜0.05].Results of multivariate analysis showed that tumor number ＞ 1,staging B of BCLC and without sequential IM after RFA were independent risk factors affecting prognosis of early HCC patients (HR=2.376,2.683,0.567,95％CI:1.080-5.229,1.530-21.112,0.335-0.962,P＜0.05).Conclusions The non-specific sequential IM of thymalfasin + IL-2 can prolong survival time of early HCC patients after RFA.Tumor number ＞ 1,staging B of BCLC and without sequential IM after RFA are independent risk factors affecting prognosis of early HCC patients.
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Objective To evaluate the clinical effect of the homemade traditional Chinese medicine bathing prescription combined with Thymalfasin and Isotretinoin treatment for Verruca Plantaris.Methods A total of 120 patients with Verruca Plantaris were randomly divided into 3 groups according to the random number table method (each group 40). The drug therapy group was given Thymalfasin and Isotretinoin to treatment; and the Chinese medicine washing group was given traditional Chinese medicine wash-out on the basis of drug therapy; and the physical therapy group was given liquid nitrogen freezing treatment on the basis of drug therapy. All the groups were treated 12 weeks and follow-up 3 months. Four weeks were a course, and a total of 3 courses. After the treatment, the symptom grading scores (number of skin lesions, the size of skin lesions, the degree of pain and the quality of life) was compared, and the clinical effect rate was evaluated and the recurrence rate was observed.Results Twelve weeks after treatment, Chinese medicine washing group total effective rate was 97.5% (39/40), drug treatment group 87.5% (35/40), and physical treatment group 90.0% (36/40). The Chinese medicine washing group total effective rate was superior to the drug therapy and physicaltherapy groups (Z values were -3.463,-3.122,P values were 0.001, 0.002). after treatment, the number of lesions scores (2.0 ± 1.1 vs. 2.8 ± 1.5, 2.7 ± 1.6,F=3.787), the size of the lesion scores (2.3 ± 1.7 vs. 3.2 ± 1.3, 3.1 ± 1.3, F=4.657), pain degree and the quality of life scores (0.4 ± 0.5 vs. 1.0 ± 1.2, 1.0 ± 1.0,F=5.353) in the Chinese medicine washing group were significantly higher than those in the drug therapy and physical therapy groups (P<0.01). After follow-up period, the recurrence rate of Chinese medicine washing group was 10.3% (4/39), the drug treatment group 25.7% (9/35), the physical therapy group 41.7% (15/36). The difference among 3 groups was statistically significant (Z=-3.188,P<0.01).Conclusions The traditional Chinese medicine combined with Thymalfasin and Isotretinoin could reduce the number of skin lesions, narrow the range of skin lesions and reduce the degree of pain, and improve the quality of life.
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Objective To observe the effect of thymalfasin on non-operative treatment of cervical cancer.Methods Seventy-eight patients with advanced cervical cancer were divided into two groups according to the digital table,each group in 39 cases.The two groups were treated with concurrent chemoradiotherapy.The observation group was given 1.6 mg of thymalfasin subcutaneously each Monday to Friday.While the control group received no additional treatment.And then the two groups were compared in terms of curative effects,adverse reactions,immune states and cellular immune functions.Results The results showed that the differences of curative effects[there were 23 patients in the control group,as compared with 31 patients in the observation group,achieved a complete response(CR),x2=3.852,P<0.05],adverse reactions(grade Ⅲ or Ⅳ radiation-induced enteritis occurred in 4 patients in the observation group,and 12 patients in the control group,x2=5.032,P<0.05;Grade Ⅲ or Ⅳ leukopenia occurred in 16 patients in the observation group,and 25 patients in the control group,x2=4.165,P<0.05;Grade Ⅲ or Ⅳ nausea,vomit occurred in 6 patients in the observation group,and 14 patients in the control group,x2=4.303,P<0.05),immune states and cellular immune functions between the two groups were significant[the counts of CD+3(t=9.236,P<0.05),CD+4(t=7.445,P<0.05),CD+4/CD+8(t=7.445,P<0.05) and NK(t=9.256,P<0.05)]were significantly higher in the observation group after treatment.Conclusion In the treatment of advanced cervical cancer,concurrent chemoradiotherapy combined with thymalfasin can improve the curative effect,reduce the side effects,improve the quality of life and enhance the immunity.
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OBJECTIVE:To study the effects of Thymalfasin for injection on the apoptosis of human lung cancer A549 cells. METHODS:After treated with 0(blank control),25,50,100,200 and 400 mg/L Thymalfasin for injection for 24,48 and 72 h, the cell proliferation inhibitory rate was analyzed with MTT and calculated. After treated with 0(blank control),50 and 100 mg/L Thymalfasin for injection for 48 h,cell apoptosis was detected by flow cytometry,and the expression of Caspase-3,Bcl-2 and Bax and the phosphorylation level of Akt were deteced by Western blot. RESULTS:Compared with blank control group,proliferation in-hibitory rate of A549 cells increased after treated with Thymalfasin for injection,in concentration and time-dependent manner(P<0.05). The apoptotic rate of A549 cells increased after treated with Thymalfasin for injection 50,100 mg/L for 48 h (P<0.05). The expression of Caspase-3 increased while the Bcl-2/Bax and phosphorylation level of Akt decreased in A549 cells after treated with Thymalfasin for injection 100 mg/L (P<0.05). CONCLUSIONS:Thymalfasin for injection can inhibit the proliferation of A549 cells by activating Caspase-3,decreasing Bcl-2/Bax ratio,inhibiting Akt signal pathway and induce the apoptosis of A549 cells.
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Objective To investigate the impact of thymalfasin on cellular immune function and lung function of patients with chronic obstructive pulmonary disease(COPD).Methods 216 patients with COPD in our hospital from March 2011 to August 2014 were randomly selected.According to the digital table,they were divided into the study group (108 cases) and control group (108 cases).The control group was given conventional treatment,while the study group was given thymalfasin on the basis of conventional treatment.Then,the cellular immune function,lung function of each index of the two groups were statistically analyzed.Results The CD3+,CD4+,CD4+/CD8+ of the study group were significantly higher (all P ＜ 0.05),the CD8+ was significantly lower(P ＜ 0.05),the FEV1,FEV1/FVC were significantly higher than the control group (P ＜ 0.05).Conclusion Thymalfasin can effectively improve the cellular immune function and lung function of patients with COPD.
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Objective To discuss the clinical effect of thymalfasin combined with radiotherapy for non -small cell lung cancer (NSCLC).Methods According to digital table,61 patients with NSCLC were randomly assigned to combined therapy group and radiotherapy group.The control group(radiotherapy group)only used radia-tion therapy.The treatment group(combination group)used thymalfasin combined with radiotherapy.The tumor lesion changes,immune indicators and the quality of life were observed.Results The effective rate of the tumor lesion changes in the treatment group was 83.9%,which in the control group was 76.7%(χ2 =0.725,P >0.05)).The levels of CD +4 ,CD +4 /CD +8 in the treatment group were (49.29 ±4.48)%,(1 .91 ±0.44),which were significantly higher than (33.1 7 ±1 .07)%,(1 .38 ±0.48)in the control group (t =1 4.69,5.34,all P <0.05).The improve-ment rate of digestive tract reaction in the treatment group was 74.2%,the body -weight ratio was 38.7%,the increased rate of KPS(karnofsky performance score)was 64.5%,while those of the control group were 23.3%, 1 3.3%,36.7% (χ2 =5.87,6.1 2,3.87,all P <0.05).Conclusion Compared with the use of radiotherapy only, thymalfasin combined with radiotherapy for NSCLC was more effective for patients in tumor lesion response,cell immu-nity and the quality of life.
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Objective To explore the efficacy and safety of thymalfasin in the treatment of severe pulmonary infection after liver transplantation.Methods Twenty seven patients who developed severe lung infection after undergoing liver transplantation in Organ Transplant Institute of the 309 th Hospital of People’s Liberation Army from January 2008 to May 2014 were enrolled in this study.According to whether the application of thymalfasin,the patients were divide into thymalfasin group (n =11)and control group (n =16).In the thymalfasin group,thymalfasin was administered via subcutaneous injection at a dose of 1.6 mg once daily for consecutive two weeks.In the control group,conventional anti-infection therapy was delivered. Ventilator time,duration of fever,the length of intensive care unit (ICU)stay and mortality were statistically compared between two groups.And the incidence of acute rejection (AR)was monitored.Results Ventilator time,duration of fever,length of ICU stay of patients in the thymalfasin group were significantly shortened compared with those in the control group (all in P <0.05).There was no significant difference in the mortality between two groups.No clinical AR was observed in either group.No thymalfasin-related adverse event was found in the thymalfasin group.Conclusions Thymalfasin can improve the curative effect to anti-infection of patients with severe pulmonary infection after liver transplantation without the incidence of AR,which is efficacious and safe in the treatment of severe pulmonary infection.
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Objective To explore the application effect of Thymalfasin for injection in the treatment of chro-nic obstructive pulmonary disease.Methods Clinical data of 80 patients with chronic obstructive pulmonary disease were retrospectively analyzed.According to different treatment methods,they were divided into the treatment group and control group,40 cases in each group.The control group was given routine treatment,the observation group were given Thymalfasin for injection with thymus method on the basis of conventional treatment.The treatment effect,improve pul-monary function,blood gas and TLC immune function of the two groups were observed.Results The total effective rate of the observation group was 95.0%,which was significantly higher than 82.5% of the control group(χ2 =5.274 5, P <0.05);After treatment in patients with pulmonary function,blood gas to improve the situation and TLC were better than those of the control group(all P <0.05);The rate of adverse reactions of the observation group was 5.0%,which was significantly lower than 20.0% of the control group(χ2 =4.114 3,P <0.05).Conclusion Thymalfasin for injection with thymus method in chronic obstructive pulmonary disease treatment has good effect,high cure rate,less adverse reaction.