Orientation and thinking of the development of laboratory medicine in the new era / 中华检验医学杂志

As an important supporting discipline of medical science, laboratory medicine plays an important role in the early diagnosis, monitoring of treatment efficacy, prognosis and risk assessment. The 21st century is an era of digital information. New detection technologies, computer science and big data of internet have brought enormous opportunities and challenges to the development of laboratory medicine. In the new era, one of the most important tasks for laboratory workers is how to make use of the revolution of information science to realize the new development of laboratory medicine. This article reviews the development of laboratory medicine, especially focusing on the orientation and future development of laboratory medicine in the new era, in order to create the prosperity of laboratory medicine development.

National survey report on the current situations and development requirements of emergency testing / 中华检验医学杂志

Objective:To investigate the current situations and development requirements of emergency testing among secondary and tertiary hospitals in China.Methods:The data were collected from secondary and tertiary hospitals via online questionnaire across 31 provinces in China from February 1 to March 1, 2021. The questionnaire involves various aspects of emergency testing, including area of emergency laboratory, staffs and equipment configuration, inspection items, Turn-around time (TAT), reagents and consumables management, pre-analysis quality control, laboratory information system, critical values management and biosafety, etc.Results:A total of 2 187 questionnaires were obtained, and 1 503 valid questionnaires from 755 secondary hospitals and 748 tertiary hospitals were finally analyzed. The research data showed that daily average number of patients visiting emergency department exceeding 300 person-time in 29.41% (220/748) tertiary hospitals, but that number was less than 100 person-time in 76.69% (579/755) secondary hospitals; daily average emergency tests exceeding 5 000 was reported in 24.47% (183/748) tertiary hospitals, and less than 2 000 was reported in 93.51% (706/755) secondary hospitals; the area of emergency laboratory was less than 100 m 2 in 68.79% (238/346) tertiary hospitals with independent emergency testing laboratory; there were no fixed staffs of emergency testing in 56.02% (842/1 503) hospitals; the biochemical/immunoassay analyzer in 8.65% (130/1 503) hospitals did not have STAT position; one hundred and twenty-six hospitals (8.38%) did not have stock in and stock out record for reagents and consumes materials; the conventional statistical analysis of unqualified specimen was not carried out in 24.62% (370/1 503) hospitals; priority on emergent specimen was not set in 58.62% (881/1 503) hospitals; whole process monitoring function was not equipped in 48.64% (731/1 503) hospitals; there was no conventional communication working mechanism with clinicians on critical value in 7.32% (110/1 503) hospitals; overall, 50.23% (755/1 503) participants did not consider that biosafety risks exist in their emergency testing laboratory. Conclusions:This survey objectively presents the current situations and future development requirements of emergency testing among secondary and tertiary hospitals in China. The survey also reflects that some important process and concepts need to be improved, and extensive attention should be paid by laboratory and hospital administrator, in the area such as communication with clinician, site construction and staff configuration, administration on the priority of emergency testing, administration on the reagent and consumable materials, laboratory informatization construction, laboratory biosafety, and so on.

How far is the future of exosomes from the laboratory to the clinic? / 中华检验医学杂志

Exosomes are nano-scale double-layer membrane structure vesicles that can be actively secreted by cells. They carry a large amount of biologically active substances and can serve as carriers for material transfer and information exchange between cells. In recent years, exosomes have become a frontier hotspot in biomedical research, and show broad application prospects in the field of laboratory diagnosis and clinical treatment of diseases. However, in general, most exosomes-related researchs are still in the laboratory research stage, and there are still many problems and challenges in separation and enrichment, technical operation specifications, and quality control. At the same time, it is urgent to carry out multi-center, large-sample clinical trials to provide evidence for exosomes from the laboratory to the clinical application.

Analysis of common mutations in non-small cell lung cancer by high-throughput sequencing / 中华检验医学杂志

Objective Next Generation Sequencing(NGS) platform was used to study the characteristics of hot gene mutations in non-small cell lung cancer (NSCLC). The distribution, type and frequency of mutation sites were systematically analyzed to evaluate the pathogenicity of mutation sites . Methods A total of 94 NSCLC tissue samples were included in this study including paraffin-embedded (FFPE) samples and fresh tissue samples, which were collected from July 2015 to April 2017 at the Qilu Hospital of Shandong University. The patient's age ranged from 35 to 82 years with a median age of 61 years. There were 63 males and 31 females. 22 hot genes in NSCLC were selected as the detection panel, including KRAS, EGFR, BRAF, PIK3CA, AKT1, ERBB2, PTEN, NRAS, STK11, MAP2K1, ALK, DDR2, CTNNB1, MET, TP53, SMAD4, FBXW7, NOTCH1, ERBB4, FGFR1 and FGFR2. Mutation detection was performed using the Ion AmpliSeq Colon and Lung Cancer Panel of the Thermo fisher's Ion Torrent sequencing platform. The sequencing data was analyzed using Ion Torrent suite v4.4.2 software. Results Among the 22 mutant genes commonly found in NSCLC, the mutation frequency of TP53 was the highest, accounting for 46.9％ of all mutations, followed by the EGFR mutation (28.1％); A total of 89 mutations were detected, including 63 hot spot mutations (reported mutations) and 26 new mutations (unreported mutations). The most frequently detected mutation was the frameshift deletion of exon 19 of EGFR, followed by the mutation of exon L858R;Analysis of the mutation in targeted drug sites of EGFR showed that the frameshift deletion of exon 19 of EGFR was the most frequently detected, followed by the mutation of exon L858R on chromosome 21. Bioinformatics software was used to analyze the pathogenicity of 26 new mutation sites. Results showed that in addition to ATK1:c. 47-12G>A and TP53: c. 214 C>G, the remaining 24 new mutation sites had at least one major impact on the gene function in three aspects, including gene conservation, amino acid sequence change and protein structure influence. Conclusion In this study, NGS was used to conduct combined detection of mutation sites of multiple hot genes, which might cover more comprehensively genetic variation and provide a basis for screening the most suitable targeted therapy groups. The pathogenicity prediction of new mutations and the changes in tumor-related signaling pathways involved provide a reference for further study of the pathogenesis of NSCLC.

The application and future development of tumor markers / 中华检验医学杂志

Human health is seriously threatened by malignancies.Early diagnosis could effectively reduce tumor mortality and improve prognosis.It remains an urgent need and difficult to evaluate the clinical application and discover more efficient diagnostic markers.The rapid development of various technology platforms, in particular the next-generation sequencing technology and translational medicine , plays crucial roles in the progress of tumor markers at the level of protein , DNA and RNA.With the development of basic researches and advances in clinical validation , and based on the prospective multicenter study , the establishment of tumor diagnostic strategy on different groups will be possible.

Establishment of detection method for serum circulating Bmi-1 mRNA and its application in diagnosis of colorectal cancer / 中华检验医学杂志

Objective To establish a direct reverse transcription real-time fluorescence quantitative polymerase chain reaction ( RT-qPCR-D ) method for detecting serum circulating B cell-specific moloney murine leukemia virus integration site-1 (Bmi-1) mRNA, and analyze the levels of serum circulating Bmi-1 mRNA in colorectal cancer patients by using of this method for exploring its diagnosis value in colorectal cancer.Methods Methodology establishment.RNA was extracted from colorectal cancer HT 29 cell line, and detection standard curves of Bmi-1, ubiquitin C ( UBC), glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) mRNAs were established , then the amplification efficiencies were calculated.Bmi-1 mRNA level was directly detected in serum and preparation buffer mixture , then the specificity of assay was evaluated by melting curve, and detection limit was observed through diluted serum samples.The serum circulating Bmi-1 mRNA levels were detected by ELISA in 158 cases with colorectal cancer , of which there were 26 cases of tumor node metastasis ( TNM)Ⅰstage, 53 cases of TNMⅡ, 47 cases of TNMⅢ, 32 cases of TNMⅣand 53 cases of controls with normal colonoscopy collected from January 2008 to January 2009 in Qilu Hospital of Shandong University.Comparisons of groups were determined by applying Mann-Whitney U test or Kruskal-Wallis test, and receiver operating characteristic ( ROC) curves were established to illustrate the diagnostic performance.Results The log values of Bmi-1, UBC and GAPDH showed good linear correlations with quantification cycle (Cq) values(R2 =0.990, 0.990, 0.991, all P 0.05).ROC curve analysis showed area under the ROC curve ( AUC) for serum circulating Bmi-1 mRNA was 0.921(95%CI=0.876-0.953), which was significantly superior to the AUC of CEA (0.745, 95%CI=0.680-0.802, Z=4.697, P0.05).Conclusions The study establishes a higher sensitive, specific for detecting serum circulating Bmi-1 mRNA. Based on this method , serum circulating Bmi-1 mRNA is found to be increased in colorectal cancer , and is superior to traditional tumor marker CEA in diagnosis of colorectal cancer, which may become a potential detection index for early detection of colorectal cancer.

The gene polymorphism of IL-1B and IL-1RN intron 2 and their relationships with the progression of HCV-related liver diseases / 中华微生物学和免疫学杂志

Objective To study the characteristics of IL-1B-31/-511 single nucleotide polymor-phisms (SNPs) and the variable number tandem repeat (VNTR) in intron 2 of the IL-1ra gene (IL-1RN) in patients with HCV-related liver diseases .Methods The concentration of IL-1βand IL-1ra in serum sam-ples was measured by ELISA assay .The SNPs of IL-1B gene (-31C/T,-511C/T) from 310 cases with HCV infection and 324 unrelated healthy controls were determined by using gene chip analysis , and the results for some randomly selected specimens were compared with those by using polymerase chain reaction -restriction fragment length polymorphism ( PCR-RFLP) assay.The VNTR polymorphism of IL-1RN intron 2 was ana-lyzed by PCR-RFLP assay.The serum level of alanine aminotransferase (ALT), an indicator of hepatocellu-lar injury, was detected by ROCHE cobas 8000 analyzer.HCV replication was measured by using specific fluorescence PCR .The genotypes of HCV were determined by direct nucleotide sequencing test .Results Compared with control group, the serum level of both IL-1β[(22.6 ±7.3) vs (13.7 ±4.2)] pg/ml and IL-1ra [(286.30 ±55.10) vs (185.55 ±48.32)] pg/ml were significantly increased in patients with HCV infection ( P0.05).The frequency of IL-1B-511TT genotype (P<0.05, OR=1.55, 95% CI =1.10-2.18) and IL-1B-511T allele (P<0.05,OR=1.31,95% CI=1.05-1.63) in patients with HCV infection were signifi-cantly higher than those in healthy controls .IL-1B-511C/T SNP showed a significant association with the outcomes of HCV infection (P<0.005).Compared with IL-1B-511CC and IL-1B-511CT, IL-1B-511TT was a major risk factor for mild and moderate Hepatitis C [ OR=2.17 ( 1.48-3.19 ) ] , severe Hepatitis C [OR=2.11(1.05-4.26)], cirrhosis [OR=2.98(1.77-4.99)] and HCC [4.33(2.16-8.67)].IL-1B-511 T allele was significantly associated with mild and moderate Hepatitis C [ 1.80 ( 1.38-2.36 ) ] , severe Hepatitis C [1.80(1.08-3.01)], cirrhosis [2.62(1.76-3.89)] and HCC [3.49(1.96-6.23)].The fre-quency of IL-1B-511T allele showed significant difference among each group (P<0.005).No association was found between any of the other polymorphisms and HCV infection .Conclusion The serum level of IL-1βand IL-1ra were significantly associated with HCV infection .IL-1B-511T allele in patients with HCV in-fection up-regulated the serum level of IL-1β.IL-1B-511TT and IL-1B-511T allele were major risk factors for mild and moderate Hepatitis C, severe Hepatitis C, cirrhosis and HCC, but IL-1B-511CC/C had oppo-site effects.

Scientific and rational application of diagnostic markers for cardiovascular disease / 中华检验医学杂志

The scientific and rational application of diagnostic marker is very important significant in the early detection and treatment of cardiovascular diseases.The emergence of cardiac-specific troponin and the development of high assays make it possible for early noninvasive diagnosis of acute myocardial infarction.The Natriuretic peptide used for the diagnosis of heart failure is a new cornerstone in the history of cardiac markers development.The future trends in new methodology development may find more potential new early markers and predictive factors,which could bring in new prospects for the early diagnosis and treatment of cardiovascular diseases,but also more challenges for clinicians and laboratory physicians.

The application of plasma soluble human leukocyte antigen in diagnosis of patients with cervical cancer and its precancerous lesion / 中华检验医学杂志

Objective To explore the application value of plasma sHLA-G in diagnosis of CIN and cervical cancer. Methods The plasma sHLA-G levels were detected by ELISA in 102 cases with cervical cancer( FIGO Ⅰ stage 32 cases, Ⅱ stage 28 cases, Ⅲ stage 25 cases and Ⅳstage 17 cases; tumor size:＜4 cm 63 cases and ≥4 cm 39 cases; squamous cell carcinoma 78 cases and adenocarcinoma 24 cases;cell differentiation:well 57 cases, moderate 29 cases and poor 16 cases; lymph nodes metastasis negative64 cases and positive 38 cases ), 72 cases with CIN( Ⅰ grade 21 cases, Ⅱ grade 25 cases and Ⅲ grade26 cases ) and 20 cases of healthy controls. The diagnostic value of sHLA-G and its correlations with clinical parameters were analyzed. Results The plasma levels of sHLA-G were 193.6( 151.3-287.4 ) kU/L in cervical cancer group, 48.3( 34.6-57.2 ) kU/L in CIN Ⅰ group, 91.3( 68.2-118.6 ) kU/L in CIN Ⅱ group, 106.4( 73.8-165.7 ) kU/L in CIN Ⅲ group and 45.2( 38.0-55.5 ) kU/L in health control group.The level of sHLA-G was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and healthy control group( U value of 8.832, 6.456, 4.017, 9.873, P ＜ 0.05,respectively ). The level of sHLA-G was significantly higher in CIN Ⅱ group and CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 4.361,4.892, 5.139, 5.485, P ＜0.05, respectively ).The levels of SCC Ag in healthy control group, CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and cervical cancer group were 0.43( 0.38-0.69 )μg/L, 0.47( 0.35-0.72 )μg/L, 0.65( 0.53-0.81 )μg/L, 0.82( 0.54-1.03 )μg/L and 1.02( 0.62-1.87 )μg/L. The level of SCC-Ag was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group and healthy control group( U value of 7.926, 4.877, 8.132,P ＜0.05, respectively ). The level of SCC-Ag was significantly higher in CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 6.574, 6.763, P ＜0.05, respectively ). The levels of CA125 in healthy control group, CIN Ⅰ group, CIN Ⅱ group, CIN Ⅲ group and cervical cancer group were 14.38 ( 6.14-21.82 ) kU/L, 15.42( 6.25-23.53 ) kU/L, 21.34( 9.82-32.58 ) kU/L, 25.69( 14.47-38.71 )kU/L and 27.72( 14.29-43.87 ) kU/L. The level of CA125 was significantly higher in cervical cancer group than that in CIN Ⅰ group, CIN Ⅱ group and healthy control group( U value of 7.564, 4.522, 7.429, P ＜0.05, respectively ). The level of CA125 was significantly higher in CIN Ⅲ group than that in CIN Ⅰ group and health control group( U value of 5.871, 5.435, P ＜0.05, respectively ). ROC curve analysis showed AUC for sHLA-G was 0.828( 95% CI:0.768-0.879 ), which was high as compared with the AUC of SCC-Ag [ 0.727( 95% CI:0.658-0.788 );Z = 2.294, P ＜ 0.05 ] and the AUC of CA125 [ 0.705( 95% CI:0.636-0.769 );Z =2.842 ,P ＜0.05 ]. There was no significant difference of diagnostic efficiency between SCC and CA125( Z =0.672, P ＞ 0.05 ). When cutoff value of sHLA-G was 109.6 kU/L, the diagnostic sensitivity,specificity, positive predictive value, negative predictive value and accuracy rate were 86.3%, 76.1%,80.0%, 83.3%, and 78.4%, respectively. The levels of sHLA-G in cervical cancer patients were significantly correlated with FIGO stages and lymphoid node metastasis ( U value of 6.085, 4.451, P ＜0.05, respectively ), while there were no significant differences between the levels of sHLA-G and age,tumor size, histological type and cell differentiation( U value of 1.274, 1.956, 1.268, 2.719, P ＞0.05,respectively ). Conclusions sHLA-G can be used for the early screening of cervical cancer and its precancerous lesion. It could also be used as an index for judging progression and lymphoid node metastasis.

Effects of heparin and low molecular weight heparin on serum level of pregnancy associated plasma protein A in patients with cerebrovascular disease / 中华检验医学杂志

Objecfive To investigate the effects of treatment for cerebrovascular disorder patients with heparin and low molecular weight heparin(LMWH) on serum PAPP-A concentrations and provide the basis for evaluating the clinical significance of PAPP-A in the following study.Methods Forty cases with cerebrovascular disease from Qilu Hospital from November 2009 to May 2010 were collected in this study.Blood samples were taken before and after drug administration.All cases were divided into four groups according to situation of medication.Group A consisted of 10 patients who received subcutaneous LMWH anticoagulation therapy, and blood samples were collected before LMWH injection, three hours after subcutaneous LMWH anticoagulation therapy in the first day, the second day and the seventh day and 24 hours after the last injection. Group B consisted of 10 patients who did not receive LMWH therapy, and blood samples were collected immediately after admission, the first day, the second day and the seventh day after admission. Group C consisted of 10 patients with percutaneous carotid intervention who received intravenous heparin at the beginning of stenting, and blood samples were collected from the arterial sheath just before angiography and heparin administration, and at 3, 5, 15, 40 and 100 min after heparin administration. Group D consisted of 10 patients who received carotid angiography but LMWH-free therapy,and blood samples were collected from the arterial sheath just before and after angiography. Serum PAPP-A concentrations were analyzed by ELISA to evaluate the differences of intra-groups and differences at different time points of inter-groups. Results In group A, PAPP-A concentrations were time dependent and elevated gradually from 12. 36 (9. 90-14. 32) mIU/L before LMWH injection to 21.80 (23.50-19.73) mIU/L at the seventh day after injection (M=38. 72, P < 0.01 ). In group C, there was a rapid increase of PAPP-A concentration from 12. 86 ( 9. 67-14. 05 ) mIU/L to 51.56 ( 44. 20-66. 00 ) mIU/L within 5 min after intravenous heparin injection (M=46. 06, P <0. 01 ). The PAPP-A concentration of one week after LMWH administration in group A was 21.80 (23.50-19.73) mIU/L, significantly higher than that in group B [11.81 (9. 21-12. 89) mIU/L] (U<0. O01, P<0.01). The PAPP-A concentration at 15 min after heparin administration in group C was 43.70 (37.70-54. 30) mIU/L, significantly higher than that after angiography in group D [14. 18 (11.25-15. 86) mIU/L] ( U<0. 001, P <0. 01 ). The peak level of blood PAPP-A after subcutaneous LMWH injection was significantly lower than that after intravenous heparin injection. The concentrations in group A and C were 21.80 ( 23.50-19. 73 ) and 51.56 (44. 20-66. 00) mIU/L respectively, and had a significant difference ( U=0. 999, P < 0. 01 ) . Conclusions Both intravenous heparin and subcutaneous LMWH administration induce an increase in serum PAPP-A concentration. The effect of drug should be considered when PAPP-A is selected as an evaluation indicator.

The roles of Tim-3 and Th-17 in children with acute idiopathic thrombocytopenic purpura / 中华检验医学杂志

Objective To investigate the expression and the role of Tim-3 and Th-17 in ITP patients and to research their clinical application. Methods Total 42 active ITP patients and 39 healthy donors were recruited in this research. The expressions of Th17 and CD4+ CD25+ Treg were measured with flow cytometer. IL-17, IFN-γ levels as well as IL-4 plasma levels were determined by ELISA. The mRNA expression of Tim-3, IFN-γ, IL-4 and T-bet were measured using RT-PCR in all samples. Results The expression of Th17 cells in ITP patients was (2.41 ± 1.43 )%, which was significantly higher than control group ( 1.08 ± 0.59)% ( t = 5.35, P < 0.05 ). But the percentage of Treg in ITP patients was ( 1.64 ±0.74)%, which was lower than control group (3.12 ±0.52)% (t = 10.33, P <0.05). The levels of IL-17 in plasma of ITP and controls were ( 14.42 ±6.37) ng/L and ( 13.91 ±4.47) ng/L respectively (t =0.42, P > 0.05). The level of IFN-γin plasma of ITP was (55.74 ± 15.25 ) ng/L, which was higher than control group (31.33 ± 12.99) ng/L (t = 7.72, P < 0.05 ). The level of IL-4 in the plasma of ITP was (7.42 ± 1.50) ng/L, which was lower than controls ( 18.17 ± 5.19) ng/L ( t = 12.87, P ＜ 0.05 ). Both IFN-γand T-bet mRNA levels were up-regulated in active ITP patients by the factor ( 8.57 ± 3.44 ) -fold and (3.34 ± 1.32)-fold than control group (t = 13.21,6.41 ,P <0.05). The decreases observed in IL-4 and Tim-3 were (0.25 ±0.15 )-fold and (0.29 ±0.15)-fold respectively in ITP patients compared with control group (t=10.02,9.61,P＜0.05 ). Conclusion The imbalance of Th17/Treg and the decrease of Tim-3 might be important determinants in the evolution of ITP.

A study on the serum proteomics in patients of colorectal cancer / 中华普通外科杂志

Objective To detect biomarker proteins in relation to metastasis by comparing serum protein profiles of primary colorectal cancer patients with or without metastasis. Methods A total of 219 serum samples were analyzed using surface enhanced laser desorption ionization time of flight mass spectrometry(SELDI-TOF-MS).The samples were divided into two groups:the training group consisting of 57 primary colorectal cancer patients、63 metastatic colorectal cancer patients and 42 healthy controls,and the test group consisting of 26 primary colorectal cancer patients and 31 metastatic colorectal cancer patients.Samples in training group were analyzed to identify serum biomarker proteins which could differentiate colorectal cancer patients with or without metastasis.The sensitivity and specificity of biomarker proteins were examined by results from blind test group. Results In the m/z region of 2000～30000,31 proteins had statistically significant difference between primary colorectal cancer patients and healthy controls.The m/z of difierentiated proteins were respectively 3240.7、9289.3、5334.4、4596.1 and 4792.4 according to P value.38 proteins had statistically significant diffefence between metastatic colorectal cancer patients and healthy controls.The m/z of differentiated proteins were respectively 3240.7、9289.3、9184.4、3191.5 and 9340.9 according to P value.Only two protein peaks(9184.4 and 9340.9)were found of statistical difference between primary eolorectal cancer patients and metastatic colorectal cancer patients.The sensitivity and specificity of the combination use of the two biomarkers were respectively 90.3% and 88.5% in the test group. Conclusion SELDI-TOF-MS was helpful to find protein biomarkers with relation to metastasis in colorectal carcinoma patients.

The diagnosis value of platelet-associated immunoglobulin G and the platelet ac tivation glycoproteins in patients with idiopathic thrombocytopenic purpura / 中华检验医学杂志

0 05). In 53 ITP patients, the positive rate of PAIgG was 64 2% (34/53), the positive rate of mean fluorescence intensity (MFI) was 71 7% (38/53), the positive rate of both PAIgG and MFI was 84 9% (45/53). The expression of GPⅡb/Ⅲa in ITP group without clinical symptoms was significantly higher than that in ITP group with clinical symptoms ( P

Detection of PMP, GPⅡb-Ⅲa, PAgT and their significances in cerebro-thrombotic diseases / 中华老年医学杂志

Objective To investigate the function and clinical significance of platelet-derived microparticles (PMP), glycoprotein(GP)Ⅱb-Ⅲa, PagT and blood-lipid in whole blood of patients with cerebro-thrombotic diseases before and after treatment. Methods The quantity of PMPs, activation ratio of GPⅡb-Ⅲa and PAgT were measured before and after treatment of cerebro-thrombotic patients by using flow cytometry and platelet adhesion instrument. Blood-lipid concentration was measured by automatic-biochemical analyzer. Results PMP, GPⅡb-Ⅲa , PAgT, TC, TG, and LDL were (223?54)/10 4 Plt, (77.98?14.22)%, (69.78?16.93) %, (5.12?0.85) mmol/L, (1.78?0.28) mmol/L, and (3.49?0.66) mmol/L respectively before treatment; and were (136?18)10 4Plt, (40.71?11.64) %, (58.12?12.51)%, (4.84?0.73) mmol/L, (1.43?0.33) mmol/L, and (3.03?0.62) mmol/L,respectively in the treatment group. These parameters were significantly decreased than that before treatment (P