ABSTRACT
Objective:To investigate the distribution of HIV-1 genotypes and drug resistance in newly diagnosed HIV-1 patients in Baoding in 2020.Methods:A self-developed method was used to amplify the pol gene sequence of HIV-1, and the sequencing results were analyzed by phylogenetic analysis and compared with the Stanford drug resistance database to determine the HIV-1 subtypes and gene mutations. Results:A total of 96 patients with HIV-1 infection were recruited in this study, and 83 pol gene sequences were successfully obtained. In the study population, 88 (91.7%) were male with an average age of 39 years and 54 (56.3%) were married. Most of the patients were infected through sexual contact (95.8%, 92/96), and 75.0% (72/96) were through homosexual transmission. Phylogenetic analysis showed that various HIV-1 subtypes were detected and among them, CRF01_AE (51.8%, 43/83), CRF07_BC (24.1%, 20/83) and B subtype (10.8%, 9/83) were the most epidemic strains. Moreover, the subtypes of newly identified recombinant strains in recent years accounted for 13.3% (11/83). Drug resistance test results showed that the pre-treatment drug resistance rate in newly diagnosed HIV-1 patients was 8.4% (7/83), and the drug resistance rates to protease inhibitor (PIs), non-nucleoside reverse transcriptase inhibitors (NNRTIs) and integrase inhibitors (INIs) were 3.6% (3/83), 1.2% (1/83) and 3.6% (3/83), respectively. Conclusions:The HIV-1 subtypes in the newly diagnosed population in Baoding in 2020 were complex and diverse. There were many unique recombinant strains and drug-resistant strains. Therefore, it was necessary to strengthen drug resistance monitoring as well as the prevention and control of HIV-1 infection in this area.
ABSTRACT
Objective:To analyze the effects of the main drug resistance mutations in the integrase (IN) region on the resistance of HIV-1 CRF01_AE strains, and compare the differences with subtype B strains.Methods:Seven IN region mutations or combined mutations (T66K, F121Y, Q148K, N155H, G118R, R263K, Q148K/N155H) were selected from the HIV drug resistance database of Stanford University in the United States, and introduced to the IN region of HIV-1 B subtype infectious clone pNL4-3 and CRF01_AE infectious clone pGX002 by seamless cloning, homologous recombination and point mutation. The mutant plasmids were transfected into 293T cells for virus packaging. The culture was expanded in MT2 cells and infectious titers were detected. Half maximal inhibitory concentrations (IC 50) of four integrase inhibitors (INSTIs), raltegravir (RAL), elvitegravir (EVG), dolutegravir (DTG) and bictegravir (BIC), against 14 mutant viruses were detected and compared with the IC 50 against the wild-type viruses. Results:B subtype and CRF01_AE plasmids carrying seven IN region mutations or combined mutations were successfully constructed, and 14 recombinant viruses were packaged with an infectious titer of 10 4-10 6 median tissue culture infective dose (TCID 50)/ml. The recombinant viruses replicated efficiently in MT2 cells. The concentrations of HIV-1 p24 antigen contained in the supernatants of cell culture reached 830-2 700 ng/ml. Five mutations or combined mutations (T66K, F121Y, Q148K, N155H, Q148K/N155H) caused CRF01_AE and B subtype strains to be highly resistant to RAL and EVG, resulting in an increase in the IC 50 by 200 times and 2 000 times or more as compared with the IC 50 against the wild-type viruses. The same mutation-caused fold changes of IC 50 of RAL and EVG against CRF01_AE were significantly lower than that of subtype B ( P<0.01). Q148K/N155H mutation caused B subtype and CRF01_AE to be highly resistant to DTG and BIC, with IC 50 increased by more than 50 times. Other mutations had little effects on the sensitivity to DTG and BIC. Conclusions:Fourteen HIV-1 strains carrying seven INSTI resistance mutations based on B subtype and CRF01_AE were constructed. Five mutations resulted in high resistance to RAL and EVG, and there was a high level of cross-resistance. Resistance to RAL and EVG caused by the same mutation was higher in B subtype than in CRF01_AE. The combined mutation of Q148K and N155H was associated with greater resistance to DTG and BIC, indicating that the genetic barrier of DTG and BIC resistance was high. DTG and BIC could effectively inhibit the strains carrying INSTI resistance mutations without obvious subtype difference.
ABSTRACT
Objective:To evaluate the diagnostic value of the Caprini risk assessment model combined with D-dimer in perioperative venous thromboembolic (VTE) of hysterectomy.Methods:The clinical data of 160 patients who had underwent hysterectomy in Lianjiang City Maternal and Child Health Hospital from February 2017 to February 2019 were retrospectively analyzed. During perioperative period, VTE occurred in 80 patients (VTE group), and 80 patients had no VTE (control group). The sensitivity, specificity, positive predictive value, negative predictive value, positive likelihood ratio, negative likelihood ratio and Youden index of Caprini risk assessment model, D-dimer level detection and Caprini risk assessment model combined with D-dimer in the diagnosis of VTE were analyzed and compared, and the diagnostic efficacy was evaluated by receiver operating characteristic (ROC) curve.Results:The positive rates of Caprini risk assessment model and D-dimer in VTE group were significantly higher than those in control group: 87.50% (70/80) vs. 17.50% (14/80) and 90.00% (72/80) vs. 41.25% (33/80), and there was statistical difference ( P<0.05). The sensitivity, specificity, negative predictive value, positive predictive value, negative likelihood ratio, positive likelihood ratio and Youden index of the Caprini risk assessment model were 87.50% (70/80), 82.50% (66/80), 86.84% (66/76), 83.33% (70/84), 0.15, 0.50 and 0.70, respectively; the indexes of D-dimer were 90.00% (72/80), 58.75% (47/80), 85.45% (47/55), 68.57% (72/105), 0.17, 2.18 and 0.49, respectively; the indexes of Caprini risk assessment model combined with D-dimer were 97.50% (78/80), 52.50% (42/80), 95.45% (42/44), 67.24% (78/116), 0.05, 2.05 and 0.50, respectively. The areas under curve of Caprini risk assessment model, D-dimer and Caprini risk assessment model combined with D-dimer were 0.888, 0.877 and 0.945 (95% CI 0.833 to 0.943, 0.820 to 0.933 and 0.908 to 0.983, P < 0.01). Conclusions:Caprini risk assessment model and D-dimer have good results in the diagnosis of perioperative VTE of hysterectomy, and Caprini risk assessment model combined with D-dimer has the highest diagnostic value.
ABSTRACT
Objective To understand the transmission patterns and risk factors of HIV-1 strain CRF01_AE subtypes in China,and to provide guidance for the implementation of precise intervention.Methods A total of 2 094 CRF01_AE pol sequences were collected in 19 provinces in China between 1996 and 2014.Phylogenetic tree was constructed by PhyML 3.0 software to select the transmission clusters.Transmission network was constructed by Cytoscape 3.6.0,which was further used for exploring of the major risk factors.Results Of the 2 094 sequences,12.18％ (255/2 094) were in clusters.A total of 82 transmission clusters were identified.The numbers of clusters and contained sequences in intra-provincial transmission (61,173) were significantly more than those in inter-provincial transmission (21,82).The ratio of transmission clustering in MSM increased over time from 2.41％ (2/83) during 1996-2008 to 23.61％ (72/305) during 2013-2014,showing a significant upward trend (x2 =27.800,df=1,P =0.000).The proportion of MSM with inter-provincial transmission clusters were higher than those with intra-provincial transmission clusters,which increased from 0.67％ (2/297) during 1996-2008 to 6.36％(30/472) during 2013-2014,showing a significant upward trend (x2=20.276,df=1,P=0.000).The transmission rate in homosexuals of the inter-transmission clusters (86.59％,71/82) was higher than that of intra-provincial transmission clusters (56.65％,98/173),and the difference was statistically significant (x2=22.792,P=0.000).The proportion of inter-provincial transmission clusters with more than 2 transmission routes (33.33％,7/21) was higher than that of intra-provincial clusters (13.11％,8/61),and the difference was statistically significant (x2=4.273,P=0.039).Results from the transmission network analysis indicated that the proportion of high risk population (degree≥4) with inter-provincial transmission clusters (51.22％,42/82) was significantly higher than that with intra-provincial transmission clusters (26.59％,46/173),and the difference was statistically significant (x2=14.932,P=0.000).Inter-provincial clusters were mainly detected in and and MSM.Conclusions Complex transmission networks were found for HIV-1 CRF01_AE strains in the mainland of China.Inter-provincial transmission clusters increased rapidly,MSM played an important role in the wide spread of the strain.More researches in transmission networks are needed to guide the precision intervention.
ABSTRACT
OBJECTIVE@#To study the effect of CYP2D610 (c.100 C>T) on plasma trough concentrations of metoprolol and its metabolite α-hydroxy metoprolol, blood pressure and heart rate in patients with coronary artery disease.@*METHODS@#The patients with coronary artery disease taking metoprolol tablets (=128) and those taking metoprolol sustained-release tablets (=126) were genotyped for CYP2D610 using Taqman real-time quantitative PCR. The trough concentrations of metoprolol and α-hydroxy metoprolol were determined with UPLC-MS/MS, and the dose-normalized concentrations (C/D) were compared among the patients with different CYP2D610 genotypes in both groups. Resting blood pressure and heart rate were recorded in all the patients when the concentration of metoprolol reached the steady state and were compared among the patients with different genotypes.@*RESULTS@#In patients taking metoprolol sustained-release tablets, the plasma trough concentration of α-hydroxy metoprolol was significantly associated with the systolic blood pressure (=0.0204). The CYP2D610 poor metabolizers showed a significant association with the C/D of metoprolol and α-hydroxy metoprolol ( < 0.01) in patients receiving metoprolol in both formulations, and in both groups, the C/D of metoprolol was significantly higher in the patients with a TT genotype than in those with a CC or CT genotype ( < 0.01); compared with those with the CT genotype, the patients with the TT genotype had a significantly lower C/D of α-hydroxy metoprolol ( < 0.01). In patients taking metoprolol sustained-release tablets, those with the CT (=0.0281) and TT (=0.0196) genotypes had lower diastolic blood pressure than patients with the CC genotypes, but the systolic blood pressure or heart rate did not differ significantly among them.@*CONCLUSIONS@#CYP2D610T allele mutation can reduce the metabolism of metoprolol, increase the C/D of metoprolol and decrease the C/D of α-metoprolol and diastolic blood pressure in patients with coronary artery disease, but CYP2D610 variation does not significantly affect systolic blood pressure or heart rate in the patients when the concentration of metoprolol reaches a steady state.
Subject(s)
Humans , Adrenergic beta-Antagonists , Chromatography, Liquid , Coronary Artery Disease , Cytochrome P-450 CYP2D6 , Genotype , Metoprolol , Tandem Mass SpectrometryABSTRACT
Objective To understand the transmission patterns and risk factors of HIV-1 strain CRF01_AE subtypes in China,and to provide guidance for the implementation of precise intervention.Methods A total of 2 094 CRF01_AE pol sequences were collected in 19 provinces in China between 1996 and 2014.Phylogenetic tree was constructed by PhyML 3.0 software to select the transmission clusters.Transmission network was constructed by Cytoscape 3.6.0,which was further used for exploring of the major risk factors.Results Of the 2 094 sequences,12.18％ (255/2 094) were in clusters.A total of 82 transmission clusters were identified.The numbers of clusters and contained sequences in intra-provincial transmission (61,173) were significantly more than those in inter-provincial transmission (21,82).The ratio of transmission clustering in MSM increased over time from 2.41％ (2/83) during 1996-2008 to 23.61％ (72/305) during 2013-2014,showing a significant upward trend (x2 =27.800,df=1,P =0.000).The proportion of MSM with inter-provincial transmission clusters were higher than those with intra-provincial transmission clusters,which increased from 0.67％ (2/297) during 1996-2008 to 6.36％(30/472) during 2013-2014,showing a significant upward trend (x2=20.276,df=1,P=0.000).The transmission rate in homosexuals of the inter-transmission clusters (86.59％,71/82) was higher than that of intra-provincial transmission clusters (56.65％,98/173),and the difference was statistically significant (x2=22.792,P=0.000).The proportion of inter-provincial transmission clusters with more than 2 transmission routes (33.33％,7/21) was higher than that of intra-provincial clusters (13.11％,8/61),and the difference was statistically significant (x2=4.273,P=0.039).Results from the transmission network analysis indicated that the proportion of high risk population (degree≥4) with inter-provincial transmission clusters (51.22％,42/82) was significantly higher than that with intra-provincial transmission clusters (26.59％,46/173),and the difference was statistically significant (x2=14.932,P=0.000).Inter-provincial clusters were mainly detected in and and MSM.Conclusions Complex transmission networks were found for HIV-1 CRF01_AE strains in the mainland of China.Inter-provincial transmission clusters increased rapidly,MSM played an important role in the wide spread of the strain.More researches in transmission networks are needed to guide the precision intervention.
ABSTRACT
Neutralizing antibodies are considered to be an important protective parameter used in HIV-l vaccine evaluation.However,the exact role that neutralizing antibodies plays in controlling the disease progression of HIV-1 infected peoples is still undetermined.In this paper,we compared the protective function of the neutralizing antibody response in the plasma from LTNP and TP against clade B and clade C pseudoviruses.No difference in the neutralizing activities between the plasma from LTNP and TP was found,which was consistent with the most recent reports.In addition,no correlations between the titer or breadth and CD4+ or viral load in HIV-1 infected individuals were found.The protective roles played by neutralizing antibodies in controlling disease progression of HIV-1 infected people need to be considered in a new viewpoint.
ABSTRACT
ObjectiveTo evaluate the sensitivity and accuracy of an in-house detecting method of HIV-1 genotypic drug resistance system. MethodsTotally 130 serum specimens from Henan and Guangxi province were collected from April 2004 to October 2008 and tested in the Military HIV Testing Center of China. ViroSeqTM v2.0 (Abbott, Switzerland), a US FDA approved HIV genotypic drug resistance detecting system was utilized as the reference method. All the specimens were detected by the novel in-house method and the reference method to validate the difference in amplifying efficiency, drug resistance mutation detection and drug resistance report. ResultsConcerning the 14 850 known drug resistance mutation sites,14 752 (99. 3％ ) mutations can be detected by both of the two methods. Rates of concordance of detection in the regions of protease inhibitors-, reverse transcriptase inhibitors- and both two classes inhibitors-resistance were99.7％ ( Kappa =0. 909 9 , P ＜0. 01 ) , 99. 0％ (Kappa=0.952 1, P＜0. 01) and99.3％ (Kappa=0. 948 8, P ＜ 0. 01 ) respectively. Drug resistance reports from these two systems showed similar results (Kappa = 0. 637 4, P ＜ 0. 01 ). The in-house detecting system identified 34 novel mutations besides the ViroSeqTM drug resistance mutation database ( ViroSeqTM software v2. 7). Two mutations, V179F and K238T,had significant effect on HIV drug resistance. ConclusionsThe in-house genotyping system is an accurate,cost-effective method and has a high concordance with commercial ViroSeqTM genotyping system. Database from the in-house assay was superior to this of the ViroSeqTM assay.
ABSTRACT
Objective To evaluate the antiretroviral therapy(ART),analyze the prevalence of resistance in rural areas,Henan,and explore the presence of minor resistant variants in pre-ART.Methods One hundred and forty-nine AIDS patients initiating ART were recruited and investigated at intervals of 6 months. Method of In-house developed by our laboratory for genotypjc resistance test was to analyze the occurrence of resistance among the failure of ART,and the allele-specific real.time PCR(ASPCR)was used to detect the minor resistant variants at the baseline samples once the resistance occurred.Results Vimlload significantly decreased among the patients who received ART(t=275,P=0.0001),but the absolute counts of CD4+T lymphocytes had no significant change(t=1.765 168,P=0.0852).Rate of resistance among the patients of treatment failure was 4.88%.The result of ASPCR in the survey of baseline showed that the minor resistant variants of M184V were detected in 7 patients and mutation K103N presented in 5 patients.Conclusion The primary drug-resistant straias in the untreated patients were found in Henan,and they might develop the dominant resistance strains and bring about the failure of ART.
ABSTRACT
Human Immunodeficiency Virus Type 1 exists in vivo as quasispecies, and one of the genome's characteristics is its diversity. During the antiretroviral therapy, drug resistance is the main obstacle to effective viral prevention. Understanding the molecular evolution process is fundamental to analyze the mechanism of drug resistance and develop a strategy to minimize resistance. Objective: The molecular evolution of drug resistance of one patient who had received reverse transcriptase inhibitors for a long time and had treatment which replaced Nevirapine with Indinavir was analyzed, with the aim of observing the drug resistance evolution pathway. Methods: The patient, XLF, was followed-up for six successive times. The viral populations were amplified and sequenced by single-genome amplification. All the sequences were submitted to the Stanford HIV Drug Resistance Database for the analysis of genotypic drug resistance. Results: 149 entire protease and 171 entire reverse transcriptase sequences were obtained from these samples, and all sequences were identified as subtype B. Before the patient received Indinavir, the viral population only had some polymorphisms in the protease sequences. After the patient began Indinavir treatment, the variants carrying polymorphisms declined while variants carrying the secondary mutation G73S gained the advantage. As therapy was prolonged, G73S was combined with M46I/L90M to form a resistance pattern M46I/G73S/L90M, which then became the dominant population. 97.9% of variants had the M46I/G73S/L90M pattern at XLF6. During the emergence of protease inhibitors resistance, reverse transcriptase inhibitors resistance maintained high levels. Conclusion: Indinavir- resistance evolution was observed by single-genome amplification. During the course of changing the regimen to incorporate Indinavir, the G73S mutation occurred and was combined with M46I/L90M.
ABSTRACT
Objective To isolate stable passage primary HIV-1 drug resistant strains and observe replication dynamics of the drug resistant isolates and evolvement tendency of the drug resistant mutations in vitro.Methods Peripheral blood mononuclear cells(PBMCs)from 15 AIDS patients receiving highly active antiretroviral therapy(HAART)were collected,and the primary HIV-1 stains were separated utilizing co-cultivated with PBMCs from normal people.HIV-1 pol genes from those strains were obtained by RT-PCR and sequenced.The drug resistant mutations were analyzed in the Stanford HIV Drug Resistance Database.Results Eight strong positive strains were isolated from 15 AIDS patients with viral loads higher than 1000 copies/ml,and two of them were drug resistant.Drug resistant mutations of the two strains were respectively K103N/K238T and M184V/K103N/Y181C/H221Y which show high-level resistance to NVP and 3TC/NVP,respectively.K103N,M184V,Y181C and H221Y exist stably in the environment without drug pressure,however,RT K238T reverted to K238.Conclusion Two drug resistant strains were successfully isolated in vitro without drug pressure.Strains with K103N shows superior fitness and can exist steadily.Strains with M184V and K103N/Y181C/H221Y can also replicate stably in vitro without drug pressure.NNRTI mutation K238T reproduces astatically,which suggests that RT 238 codon might revert gradually to wild genotype.
ABSTRACT
Objective To clarify the serological characteristics and predictive value of HIV antibody indeterminant and to evaluate the efficiency of 3 assays to discriminate HIV antibody indeterminant.Methods Three hundred and ninety-four HIV antibody indeterminant serum samples were collected and the Western blot pattern were analyzed.Ninety-seven HIV antibody indeterminant individuals were followed up,and the development of HIV antibody were observed.The initial serum samples of 67 followed individuals were tested by viral load,line immunoblot assay and ELISA for HIV-1 p24,with the golden standard of follow up,the efficiency of 3 kinds of assay to discriminate HIV antibody indeterminant were evaluated.Results There were 38 patterns among 394 HIV antibody indeterminant,the proportions of env,pol and gag indeterminant were 37.54%,4.04%and 58.37% respectively.Five HIV antibody indeterminant cases were converted to HIV antibody positive among 97 followed individuals,they were all env indeterminant and HIV antibody developed rapidly.HIV viral load was an ideal assay to discriminate HIV antibody indeterminant with best sensitivity.Conclusion The indeterminant of gag were most common,but were unspecific reaction.Env indeterminant were with the greatest predictive value of HIV infection,especially the gp160p24 and gp160.Viral load assay can be applied to discriminate HIV antibody indeterminant.
ABSTRACT
Objective To develop and evaluate the allele-specific real-time PCR(ASPCR) assay for the detection of minor HIV-1 variants. Methods We developed and evaluated the ASPCR assay, using the K103N mutation site as a model system. We constructed plasmids as standards and designed specific and non-specific primers to discriminate the wild-type and mutant plasmids in the real-time PCR using SYBR green as fluorescence reporter. And then we evaluated the sensitivity, accuracy, reproducibility of ASPCR assay and detected the control samples. Results The specific primer can discriminate the wild-type and mutant plasmids including resistant mutation successfully. The sensitivity of ASPCR assay can achieve less than 0.01% and the accuracy of this method is down to 0.1%. The Intra-assay coefficient of variation is less than 0.7 and the Inter-assay coefficient of variation is less than 1.6. Conclusion ASPCR is a sensitive, accurate and rapid method to detect the minor HIV-1 variants which have resistant mutations and it can be used widely in HIV research. ASPCR also can provide earlier and more resistant information to the clinical therapy.
ABSTRACT
Objective To elucidate the molecular evolutional characteristics of HIV-1 non-nucleoside reverse transcriptase inhibitor (NNRTI) drug resistance-associated mutations in AIDS patients receiving highly active antiretroviral therapy (HAART).Methods Four AIDS patients receiving HAART with good adherence within a HlV-1 drug resistance cohort from a rural region in central China were selected,who possessed susceptible virus at the beginning of treatment and gradually came to produce resistance to NNRTIs during the process of antiretroviral therapy (ART),reverse transcriptase (RT) genes from each patient's peripheral blood samples (from 3 to 30 months after withdrawal) were cloned and sequenced in succession.Results To sequenced total 855 clones and obtained the HIV-1 NNRTI drug resistance-asseciated mutations patterns of the four patients: (1)G190A often appeared with F227 L and had the tendency of accumulating P236V during the process of treatmenL (2)Y188C always presented alone and sometimes it concured with P236V.(3) YI81C frequently concured with VI79D or KIO3N and the combination varies from patient to patient.(4)K103N often combined with Y181C or M230L Conclusions The molecular evolutional characteristics of HIV-1 NNRTI drug resistance-asseciated mutations in the 4 AIDS patients are summarized.They showed different pathways on HIV-1 NNRTI drug resistance-associated mutations and those mutations detected early tend to be predominant strains.
ABSTRACT
Objective To characterize 5 gpl20 sequences from mainly circulating clades in China and expression of their gp120 glycoproteins. Methods gp120 genes were amplified from the PBMCs of 5 HIV-1 infected individuals in different provinces using nest PCR and their DNA sequences were determined. Sequence characteristics were analyzed and gp120 genes were sub-cloned into the mammalian expression vec-tor to produce gp120 glycoproteins. Results Sequence characteristics indicated these sequences belong to the clade Thai-B, CRF_BC and CRF_AE, respectively. There were some conservative N-linked glycosyla-tion sites and primary Furin protease cleavage motifs in the same positions within gp120 amino acid se-quences although these gp120 sequences were categorized into different clades. In comparison with referen-tial strains, amino acids deletions were found in the V1, V2, V4, V5 regions except for the V3 loop; above all, V3 tip motifs of Thai-B exhibited the more polymorphic forms than those of CRF_BC and CRF_AE. These 5 gp120 sequences were cloned into the eukaryotic expression vector and gpl20 glycoproteins were produced successfully. Conclusion Hyper-variable nature of Env should be considered while designing HIV-1 vaccine or test reagent, and gpl20 expression in vitro is helpful to further research on the Env patho-genesis and vaccine development against the mainly circulating HIV-1 isolates in China.
ABSTRACT
Objective To evaluate the feasibility and safety of vaginal enlarged amputation of cervix to treat patients with cervical cancer of stage Ⅰ a1 and cervical intraepithelial neoplasia grade Ⅲ(CIN Ⅲ)who were unfit for conization surgery.Methods From July 2002 to May 2007,patients with cervical cancer at stage Ⅰ a1,diagnosed by pathology after loop electrosurgical excision procedure(LEEP),large area CIN Ⅲ(the area of lesion≥3/4 on colposcopy),CIN Ⅲ coexisted with vaginal intraepithelial neoplasia (VAIN)in the superior segment of vagina,CIN Ⅱ-Ⅲ recurrence or with residual lesion,positive margin after conization of cervix,who wanted to preserve fertility and(or)corpus uteri were selected to receive vaginal enlarged amputation of cervix.Results Forty-eight eases including 5 with cervical cancer in stage Ⅰ a1,38 with large area CIN Ⅲ(9 with gland involvement),2 with residual lesion and 2 with positive margin after LEEP,1 recurrence after cold knife conization,received the procedure successfully.The median age was 34 years(range 27-40),median operation time was 60 minutes(range 30-100),median blood loss was 40 ml(range 5-300),and median hospital stay was 10 days(range 7-17).After follow-up 1-39 months,no patient had postoperative complications and recurrence,and all patients resumed normal menstrual cycle and sexual life.Condusion Vaginal enlarged amputation of cervix appears to be a safe and feasible procedure for patients with cervical cancer at stage Ⅰ a1 and CIN Ⅲ who are unfit for conization surgery.