Perianal disease in inflammatory bowel disease:Broadening treatment and surveillance strategies for anal cancer

The perianal disease affects up to one-third of individuals with Crohn's disease(CD),causing disabling symptoms and significant impairment in quality of life,particularly for those with perianal fistulising CD(PFCD).The collaborative effort between gastroenterologists and surgeons is essential for addressing PFCD to achieve fistula closure and promote luminal healing.Limited fistula healing rates with conventional therapies have prompted the emergence of new biological agents,endoscopic procedures and surgical techniques that show promising results.Among these,mesenchymal stem cells injection is a particularly hopeful therapy.In addition to the burden of fistulas,individuals with perianal CD may face an increased risk of developing anal cancer.This underscores the importance of surveillance programmes and timely interventions to prevent late diagnoses and poor outcomes.Currently,there is no established formal anal screening programme.In this review,we provide an overview of the current state of the art in managing PFCD,including novel medical,endoscopic and surgical approaches.The discussion also focuses on the relevance of establishing an anal cancer screening programme in CD,intending to propose a risk-based surveillance algorithm.The validation of this surveillance programme would be a significant step forward in improving patient care and outcomes.

Anal Pathologies: What Management at the Reference Health Center of Bamako’s Commune V?

Anal pathologies are conditions affecting the anal margin and the anal canal. The aim of our study was to update data on anal diseases. Non-specific, it can reveal an emergency, a serious illness, a chronic disease or a benign condition. It was a prospective, descriptive and analytical study from April 2022 to March 2023, carried out in the hepato-gastroenterology unit of the Centre de Santé de Référence de la commune V du District de Bamako in Mali. Out of seven hundred and forty-nine (749) patients seen in consultation, 98 had anal pathology, i.e. a prevalence of 13.08%. The mean age was 40 ± 15.15 years and the sex ratio was 1.5. Hemorrhoidal disease and anal fissure were the most frequent pathologies in 66.4% and 24.4% of cases respectively. Medical treatment was initiated in 91.9% of patients with hemorrhoidal disease and 91.7% with anal fissure. Anal pathologies are common in young male patients. In our context, they are dominated by benign conditions.

Aqueous Extract of Ceiba pentandra Stimulates the Production of Fetal Hemoglobin in Sickle Cell Patients

Subsequent studies have demonstrated the reversed activity of the aqueous extract of Ceiba pentandra on the deformity of sickled red blood cells in hypoxia conditions. The observation which related to an in vitro study had given rise to hopes as to the management of sickle cell disease (SCD) by the use of this plant species. In this paper, the authors aimed to investigate the effect of the aqueous extract of C. pentandra on the production of fetal hemoglobin in SCD patients. The work carried out hemoglobin electrophoresis, for a period of six months, on blood samples from SCD patients who voluntarily undergone routine treatment, based on the medicinal recipe prepared from the bark of the trunk and branches of C. pentandra, in a hospital center of herbal medicines located in Kinshasa. The medicinal recipe called BEAT-SS is a patented product of the hospital center named Centre de Phytothérapie Moderne NIECA. Blood samples from patients under treatment were taken to evaluate the behavior of different forms of hemoglobin (hemoglobin S, hemoglobin F and hemoglobin A2). Agarose gel electrophoresis with integrated reading was used for the separation of the different forms of hemoglobin, as well as their dosage on each sample of sickle blood. A reduction in the proportion of hemoglobin S and an increase in the proportion of fetal hemoglobin were found in all sickle cell patients during the treatment period. This observation could affirm that the management of sickle cell patients using the recipe prepared from the aqueous extract of C. pentandra could increase the level of fetal hemoglobin in these patients.

A comparative study of acupuncture combined with rehabilitation gymnastics on postoperative anal function of lower rectal cancer

BACKGROUND From the anal function,inflammatory response and other indicators,acupuncture combined with rehabilitation gymnastics was applied to patients with cancer undergoing low resection,aiming to improve the prognosis of patients.AIM To explore the effects of acupuncture combined with rehabilitation gymnastics on anal function after lower rectal cancer surgery.METHODS From January 2020 to December 2022,128 patients who underwent rectal cancer surgery in the Department of Oncology of Hebei Provincial Hospital of Traditional Chinese Medicine Hospital were selected and divided into two groups using the random number table method,with 64 patients in each group.Patients in the control group were not treated with acupuncture or rehabilitation gymnastics and served as blank controls.Patients in the study group were treated with acupuncture and rehabilitation gymnastics from the 7th postoperative day.The anal incontinence scores,changes in serum interleukin-4,interleukin-6,and interleukin-10 Levels,and serum motilin,5-hydroxytryptamine,and vasoactive intestinal peptide levels were compared.RESULTS There were no significant differences in serum interleukin-4,interleukin-6,and interleukin-10 Levels between the groups before treatment(P>0.05).After treatment,these levels were better than those of the control group(P<0.05).There was no significant difference in the anal incontinence scores between the groups before and 7 d after surgery(P>0.05).Anal incontinence scores in the study group were lower than those in the control group at 14 d,21 d,and 28 d postoperatively(P<0.05).There were no significant differences in serum motilin,5-hydroxytryptamine,or vasoactive intestinal peptide levels between the groups before treatment(P>0.05).After treatment,these levels were higher in the study group than in the control group,and vasoactive intestinal peptide level was lower in the study group than in the control group(P<0.05).CONCLUSION Acupuncture combined with rehabilitation gymnastics can promote the recovery of anal function and reduce the inflammatory response in patients with lower rectal cancer after surgery.

Establishment of a surveillance program for anal cancer in Crohn's disease

This letter discusses the incidence of anal cancer among Crohn's disease(CD)-related malignancies.Patients with CD have been demonstrated to be at a higher risk of developing small bowel and colorectal cancers than healthy individuals.Although CD-associated anal cancer is relatively rare,patients with CD accom-panied by anal or perianal lesions are at increased risk of anal cancer.Addi-tionally,compared to ulcerative colitis,which is also an inflammatory disease,CD-related anal cancer is often detected at an advanced stage owing to the complexity of CD and is associated with a poor prognosis with frequent local recurrences.Therefore,the detection of early-stage cancer is crucial for improving the prognosis.However,the surveillance methods recommended for CD-related malignancies in the United States and Europe are similar to those for ulcerative colitis.They are not appropriate for detecting CD-related malignancies in the recto-anal region.Therefore,there is an urgent need for surveillance programs aimed at the early detection of malignant anorectal lesions in patients with CD.

Structured magnetic resonance imaging and endoanal ultrasound anal fistulas reporting template(SMART):An interdisciplinary Delphi consensus

BACKGROUND There is still considerable heterogeneity regarding which features of cryptoglandular anal fistula on magnetic resonance imaging(MRI)and endoanal ultrasound(EAUS)are relevant to surgical decision-making.As a con-sequence,the quality and completeness of the report are highly dependent on the training and experience of the examiners.AIM To develop a structured MRI and EAUS template(SMART)reporting the minimum dataset of information for the treatment of anal fistulas.METHODS This modified Delphi survey based on the RAND-UCLA appropriateness for consensus-building was conducted between May and August 2023.One hundred and fifty-one articles selected from a systematic review of the lite-rature formed the database to generate the evidence-based statements for the Delphi study.Fourteen questions were anonymously voted by an interdisciplinary multidisciplinary group for a maximum of three iterative rounds.The degree of agreement was scored on a numeric 0–10 scale.Group consensus was defined as a score≥8 for≥80%of the panelists.RESULTS Eleven scientific societies(3 radiological and 8 surgical)endorsed the study.After three rounds of voting,the experts(69 colorectal surgeons,23 radiologists,2 anatomists,and 1 gastroenterologist)achieved consensus for 12 of 14 statements(85.7%).Based on the results of the Delphi process,the six following features of anal fistulas were included in the SMART:Primary tract,secondary extension,internal opening,presence of collection,coexisting le-sions,and sphincters morphology.CONCLUSION A structured template,SMART,was developed to standardize imaging reporting of fistula-in-ano in a simple,systematic,time-efficient way,providing the minimum dataset of information and visual diagram useful to refer-ring physicians.

Anal pruritus:Don’t look away

Anal pruritus is a common anorectal symptom that can significantly impair a patient’s quality of life,including their mental health.It can be one of the most difficult proctological conditions to treat.Patients often delay seeking medical attention,since it is an embarrassing but non-life-threatening situation.Pruritus ani can be associated with idiopathic and secondary causes,such as anorectal diseases,cancer(anal or colorectal),dermatological and sexually transmitted diseases,fungal infections and systemic diseases.If patients are referred for a colonoscopy,this can sometimes provide the first opportunity to evaluate the perianal area.Classifications of anal pruritus are based on the abnormalities of the perianal skin,one of the most commonly used being the Washington classi-fication.A proper digital anorectal examination is important,as well as an anoscopy to help to exclude anorectal diseases or suspicious masses.Endoscopists should be aware of the common etiologies,and classification of the perianal area abnormalities should be provided in the colonoscopy report.Information on treatment possibilities and follow-up can also be provided.The treatment normally consists of a triple approach:proper hygiene,elimination of irritants,and skin care and protection.Several topical therapies have been described as possible treatments,including steroids,capsaicin,tacrolimus and methylene blue intradermal injections.

High complex anal fistula managed by the modified transanal opening of the intersphincteric space via the inter-sphincteric approach:A case report

BACKGROUND High complex anal fistulas are epithelialized tunnels,with the main fistula piercing above the deep external sphincter and the internal opening approaching the dentate line.Conventional surgical procedures for high complex anal fistulas remove most of the external sphincter and damage the anorectal ring.Postoperative loss of anal function can cause physical and mental damage.Transanal opening of the intersphincteric space(TROPIS)is an effective procedure that completely preserves the external anal sphincter.However,its clinical application is limited by challenges in the localization of the internal opening of a fistula and the high risk of complications.On the basis of our clinical experience,we modified the TROPIS procedure for the treatment of treating high complex anal fistulas.CASE SUMMARY A patient with a high complex anal fistula located above the anorectal ring underwent modified TROPIS,which involved sepsis drainage and identification of the internal opening in the intersphincteric space.The patient with the high complex anal fistula recovered well postoperatively,without any postoperative complications or anal dysfunction.Anal function returned to normal after 17 months of follow-up.CONCLUSION The modified TROPIS procedure is the most minimally invasive surgery for anal fistulas that minimally impairs anal function.It allows the complete removal of infected anal glands and reduces the risk of postoperative complications.Modified TROPIS via the intersphincteric approach is an alternative sphincter-preserving treatment for high complex anal fistulas.

Associations of serum D-dimer and glycosylated hemoglobin levels with third-trimester fetal growth restriction in gestational diabetes mellitus

BACKGROUND Gestational diabetes mellitus(GDM)is a special type of diabetes that commonly occurs in women during pregnancy and involves impaired glucose tolerance and abnormal glucose metabolism;GDM is diagnosed for the first time during pregnancy and can affect fetal growth and development.AIM To investigate the associations of serum D-dimer(D-D)and glycosylated hemoglobin(HbA1c)levels with third-trimester fetal growth restriction(FGR)in GDM patients.METHODS The clinical data of 164 pregnant women who were diagnosed with GDM and delivered at the Obstetrics and Gynecology Hospital of Fudan University from January 2021 to January 2023 were analyzed retrospectively.Among these women,63 whose fetuses had FGR were included in the FGR group,and 101 women whose fetuses had normal body weights were included in the normal body weight group(normal group).Fasting venous blood samples were collected from the elbow at 28-30 wk gestation and 1-3 d before delivery to measure serum D-D and HbA1c levels for comparative analysis.The diagnostic value of serum D-D and HbA1c levels for FGR was evaluated by receiver operating characteristic analysis,and the influencing factors of third-trimester FGR in GDM patients were analyzed by logistic regression.RESULTS Serum fasting blood glucose,fasting insulin,D-D and HbA1c levels were significantly greater in the FGR group than in the normal group,while the homeostasis model assessment of insulin resistance values were lower(P<0.05).Regarding the diagnosis of FGR based on serum D-D and HbA1c levels,the areas under the curves(AUCs)were 0.826 and 0.848,the cutoff values were 3.04 mg/L and 5.80%,the sensitivities were 81.0%and 79.4%,and the specificities were 88.1%and 87.1%,respectively.The AUC of serum D-D plus HbA1c levels for diagnosing FGR was 0.928,and the sensitivity and specificity were 84.1%and 91.1%,respectively.High D-D and HbA1c levels were risk factors for third-trimester FGR in GDM patients(P<0.05).CONCLUSION D-D and HbA1c levels can indicate the occurrence of FGR in GDM patients in the third trimester of pregnancy to some extent,and their combination can be used as an important index for the early prediction of FGR.

Hemoglobin loss method calculates blood loss during pancreaticoduodenectomy and predicts bleeding-related risk factors

BACKGROUND The common clinical method to evaluate blood loss during pancreaticoduoden-ectomy(PD)is visual inspection,but most scholars believe that this method is extremely subjective and inaccurate.Currently,there is no accurate,objective me-thod to evaluate the amount of blood loss in PD patients.We retrospectively analyzed the clinical data of 341 patients who underwent PD in Shandong Provincial Hospital from March 2017 to February 2019.According to different surgical methods,they were divided into an open PD(OPD)group and a laparoscopic PD(LPD)group.The differences and correlations between the in-traoperative estimation of blood loss(IEBL)obtained by visual inspection and the intraoperative calculation of blood loss(ICBL)obtained using the Hb loss method were analyzed.ICBL,IEBL and perioperative calculation of blood loss(PCBL)were compared between the two groups,and single-factor regression analysis was performed.RESULTS There was no statistically significant difference in the preoperative general patient information between the two groups(P>0.05).PD had an ICBL of 743.2(393.0,1173.1)mL and an IEBL of 100.0(50.0,300.0)mL(P<0.001).There was also a certain correlation between the two(r=0.312,P<0.001).Single-factor analysis of ICBL showed that a history of diabetes[95%confidence interval(CI):53.82-549.62;P=0.017]was an independent risk factor for ICBL.In addition,the single-factor analysis of PCBL showed that body mass index(BMI)(95%CI:0.62-76.75;P=0.046)and preoperative total bilirubin>200μmol/L(95%CI:7.09-644.26;P=0.045)were independent risk factors for PCBL.The ICBLs of the LPD group and OPD group were 767.7(435.4,1249.0)mL and 663.8(347.7,1138.2)mL,respectively(P>0.05).The IEBL of the LPD group 200.0(50.0,200.0)mL was slightly greater than that of the OPD group 100.0(50.0,300.0)mL(P>0.05).PCBL was greater in the LPD group than the OPD group[1061.6(612.3,1632.3)mL vs 806.1(375.9,1347.6)mL](P<0.05).CONCLUSION The ICBL in patients who underwent PD was greater than the IEBL,but there is a certain correlation between the two.The Hb loss method can be used to evaluate intraoperative blood loss.A history of diabetes,preoperative bilirubin>200μmol/L and high BMI increase the patient's risk of bleeding.

Evaluating the clinical application and effect of acupuncture therapy in anal function rehabilitation after low-tension rectal cancer surgery

BACKGROUND According to the indexes of serum and anal function,acupuncture therapy was applied to patients with low rectal cancer in order to avoid the occurrence of anal incontinence and reduce complications.AIM To explore the clinical application and evaluate the effect of acupuncture therapy for anal function rehabilitation after low-tension rectal cancer surgery.METHODS From the anorectal surgery cases,we selected 120 patients who underwent colorectal cancer surgery between January 2020 and December 2022 and randomly divided them into a control group(n=60),observation group(n=60),and control group after surgery for lifestyle intervention(including smoking cessation and exercise),dietary factor adjustment,anal movement,and oral loperamide treatment.The serum levels of motilin,5-hydroxytryptamine,and vasoactive intestinal peptide(VIP),Wexner score for anal incontinence,and incidence of complications were compared between groups.RESULTS After treatment,the VIP and 5-hydroxytryptamine levels in the observation group were lower than those in the control group(P<0.05).The motilin level was higher than that in the control group(P<0.05).Postoperative anal incontinence was better in the observation group than in the control group(P<0.05).The incidence of complications in the observation group was 6.67%,which was significantly lower than that in the control group(21.67%;P<0.05).CONCLUSION Acupuncture therapy has a positive effect on the rehabilitation of anal function after low-tension rectal cancer surgery;it can effectively help to improve the serum indices of patients,avoid the occurrence of anal incontinence,and reduce the incidence of complications.Popularizing and applying it will be valuable.

Relationship between hemoglobin glycation index and risk of hypoglycemia in type 2 diabetes with time-in-range in target

BACKGROUND In patients with type 2 diabetes mellitus(T2DM),the risk of hypoglycemia also occurs in at a time-in-range(TIR)of>70%.The hemoglobin glycation index(HGI)is considered the best single factor for predicting hypoglycemia,and offers new perspectives for the individualized treatment of patients with well-controlled blood glucose levels that are easily ignored in clinical settings.All participants underwent a 7-days continuous glucose monitoring(CGM)using a retrospective CGM system.We obtained glycemic variability indices using the CGM system.We defined HGI as laboratory hemoglobin A1c minus the glucose management indicator.Patients were categorized into low HGI(HGI<0.5)and high HGI groups(HGI≥0.5)according to HGI median(0.5).Logistic regression and receiver operating characteristic curve analyses were used to determine the risk factors for hypoglycemia.RESULTS We included 129 subjects with T2DM(54.84±12.56 years,46%male)in the study.Median TIR score was 90%.The high HGI group exhibited lower TIR and greater time below range with higher hemoglobin A1c than the low HGI group;this suggests more glycemic excursions and an increased incidence of hypoglycemia in the high HGI group.Multivariate analyses revealed that mean blood glucose,standard deviation of blood glucose and HGI were independent risk factors for hypoglycemia.Receiver operating characteristic curve analysis indicated that the HGI was the best predictor of hypoglycemia.In addition,the optimal cut-off points for HGI,mean blood glucose,and standard deviation of blood glucose in predicting hypoglycemia were 0.5%,7.2 mmol/L and 1.4 mmol/L respectively.CONCLUSION High HGI was significantly associated with greater glycemic excursions and increased hypoglycemia in patients with TIR>70%.Our findings indicate that HGI is a reliable predictor of hypoglycemia in this population.

Cut-off value of glycated hemoglobin A1c for detecting diabetic retinopathy in the Chinese population

BACKGROUND Glycated hemoglobin A1c(HbA1c)is considered the most suitable for diabetes mellitus diagnosis due to its accuracy and convenience.However,the effect of HbA1c on diabetic retinopathy(DR)in the Han and Korean populations in Jilin,China,remains inconclusive.AIM To determine the best cut-off of HbA1c for diagnosing DR among the Chinese.METHODS This cross-sectional study included 1933 participants from the Yanbian area of Jilin Province,China.Trained investigators employed a questionnaire-based survey,physical examination,laboratory tests,and fundus photography for the investigation.The best cut-off value for HbA1c was established via the receiver operating characteristic curve.The factors associated with HbA1c-associated risk factors were determined via linear regression.RESULTS The analysis included 887 eligible Chinese Han and Korean participants,591 of whom were assigned randomly to the training set and 296 to the validation set.The prevalence of DR was 3.27% in the total population.HbA1c of 6.2% was the best cut-off value in the training set,while it was 5.9% in the validation set.In both Chinese Han and Korean populations,an HbA1c level of 6.2% was the best cut-off value.The optimal cut-off values of fasting blood glucose(FBG)≥7 mmol/L and<7 mmol/L were 8.1% and 6.2% respectively in Han populations,while those in Korean populations were 6.9%and 5.3%,respectively.Age,body mass index,and FBG were determined as the risk factors impacting HbA1c levels.CONCLUSION HbA1c may serve as a useful diagnostic indicator for DR.An HbA1c level of 6.2% may be an appropriate cut-off value for DR detection in the Chinese population.

Anal carcinoma-exploring the epidemiology,risk factors,pathophysiology,diagnosis,and treatment

Anal carcinoma is a relatively rare tumor that accounts for approximately 2%of gastrointestinal malignancies and less than 7%of anorectal cancers.Most anal tumors originate between the anorectal junction and the anal verge.Risk factors for the disease include human papillomavirus infection,human immunodeficiency virus,tobacco use,immunosuppression,female sex,and older age.The pathogenesis of anal carcinoma is believed to be linked to human papillomavirusrelated inflammation,leading to dysplasia and progression to cancer.Squamous cell carcinoma is the most common type of anal tumor,with an annual incidence of approximately 1 to 2 per 100000 persons.Treatment regarding anal cancer has emerged over time.However,chemoradiation therapy remains the mainstay approach for early localized disease.Patients with metastatic disease are treated with systemic therapy,and salvage surgery is reserved for disease recurrence following chemoradiation.This article aims to provide background information on the epidemiology,risk factors,pathology,diagnosis,and current trends in the management of anal cancer.Future directions are briefly discussed.

Clinical Study on the Treatment of Low Anal Fistula in Infants and Young Children by Anal Gland Excision and Virtual Hanging Procedure

Objective:To compare the efficacy of anal adenectomy with virtual hanging wire and anal fistulotomy in the treatment of low anal fistula in infants and children.Methods:60 children with low anal fistula who were admitted to our hospital from October 2021 to March 2022 and met the inclusion criteria were randomly divided into two groups of 30 cases each;the treatment group was treated with anal adenectomy and virtual hanging wire surgery,and the control group was treated with anal fistula resection.The clinical efficacy after treatment was compared.Results:The total effective rate of both groups was 96.67%and the difference between the two groups was not statistically significant(P>0.05).The postoperative pain score of the treatment group was lower than that of the control group(P<0.05).The length of hospitalization and healing time of the treatment group was lower than that of the control group(P<0.05).The anal function of the patients in both groups was normal,and there was no adverse reaction.Conclusion:Anal gland excision and virtual hanging surgery for the treatment of low anal fistula in infants and children have the advantages of mild pain,reduced length of hospitalization,short healing time,and better patient experience as compared to anal fistula excision.

A case study illustrating the use of an integrated approach to treat secondary Anal Fistula Post I&D of Perianal Abscess

Background:Anal fistula is a long-term disease characterized by a tubular structure with one end opening in the anorectal canal and the other end opening on the surface of the perineum or perianal skin with chronic pus drainage.It is linked to Bhagandar in Ayurveda,and in Sushruta Samhita,Acharya has mentioned 5 forms of Bhagandar.The boil in the present case was Shukla,sthira i.e.hard and firm,with Picchila strava and Kandu resembling the features of Parisraavi bhagandar.Aim and objective:The current case was diagnosed as Parisravi bhagandar,which resembles trans-sphincteric or intersphincteric fistula in modern ano.In Ayurveda,the management of Parisraavi Bhagandar,Shastra,kshara,and Agnikarma is advised and the use of Ksharasutra,which contributes to complete cutting and healing of the track without reoccurrence,similarly Modern surgeon depends on surgery i.e radical excision of the track,ligation with Seton,and use of chemical irritants like urethane,silver nitrates,etc.A cutting seton(tight)gently slices the confined muscle to close the fistula with the least interruption to continence.This operation is especially advised when a one-stage fistulotomy poses a considerable risk of incontinence.Material and methods:The method performed here was Core Partial Fistulectomy followed by Ksharasutra application till complete healing of the wound.Discussion and conclusion:This case study provides the successful management of Parisraavi Bhagandara(high anal,trans-sphincteric fistula in ano)in 61-year-old male patient with an integrated surgical&Ayurvedic management approach.

Epidemiological Survey of Hemoglobinopathies Based on Next-Generation Sequencing Platform in Hunan Province,China

Objective This study was aimed at investigating the carrier rate of,and molecular variation in,α-andβ-globin gene mutations in Hunan Province.Methods We recruited 25,946 individuals attending premarital screening from 42 districts and counties in all 14 cities of Hunan Province.Hematological screening was performed,and molecular parameters were assessed.Results The overall carrier rate of thalassemia was 7.1%,including 4.83%forα-thalassemia,2.15%forβ-thalassemia,and 0.12%for bothα-andβ-thalassemia.The highest carrier rate of thalassemia was in Yongzhou(14.57%).The most abundant genotype ofα-thalassemia andβ-thalassemia was-α^(3.7)/αα(50.23%)andβ^(IVS-Ⅱ-654)/β^(N)(28.23%),respectively.Fourα-globin mutations[CD108(ACC>AAC),CAP+29(G>C),Hb Agrinio and Hb Cervantes]and sixβ-globin mutations[CAP+8(C>T),IVS-Ⅱ-848(C>T),-56(G>C),beta nt-77(G>C),codon 20/21(-TGGA)and Hb Knossos]had not previously been identified in China.Furthermore,this study provides the first report of the carrier rates of abnormal hemoglobin variants andα-globin triplication in Hunan Province,which were 0.49%and 1.99%,respectively.Conclusion Our study demonstrates the high complexity and diversity of thalassemia gene mutations in the Hunan population.The results should facilitate genetic counselling and the prevention of severe thalassemia in this region.

MALDI Mass Spectrometry Analysis of Human Hemoglobin

Matrix-assisted laser desorption/ionisation(MALDI) mass spectra in matrices DHBs and ferulic acid have been taken and used to determine the molecular weight of one of the variants of human hemoglobin. A single laser operating at 355 nm was used to irradiate the sample droplet spots on a target for both desorption and ionisation. Ions were analyzed and detected by a reflectron time-of-flight mass spectrometer system. The mass values of α-chain and β-chain without heme group for a kind of human hemoglobin molecules were determined, which are 15275 Da and 16060Da respectively, with the accuracy of 0. 03%. The various factors influencing the resolution of mass spectrum were discussed.

Analysis of styrene oxide-hemoglobin adducts at different binding sites

An assay was described for measures of adducts of styrene oxide (SO) with molecules of hemoglobin(Hb) which have been modified at residues containing either carboxylic acid or sulfhydryl groups. The method employs two steps. In the first, SO carboxylic acid adducts are hydrolyzed in basic medium to liberate styrene glycol(SG). Then, the remaining residues are reacted with a reductive catalyst (Raney nickel) to liberate the two positional isomers of SO cysteine adducts, i.e., 1 phenylethanol (1 PE) and 2 phenylethanol (2 PE). The liberated products are derivatized with pentafluorobenzoyl chloride and measured by GC NICI MS. The assay was evaluated with SO adducts of Hb which had been produced in the blood of Sprague Dawley rats following (a) in vitro modification(0—300m mol/L SO) or (b) i.p administration of either SO(0—1 mmol/kg) or styrene (0—3 mmol/kg). Levels of each of the three analytes (SG, 1 PE, and 2 PE) increased with dose in hemoglobin. The ratios of cysteine adducts to carboxylic acid adducts varied significantly among experiments. Ratios of 2 PE to 1 PE were much more consistent (2 PE/1 PE=6.9 (SE=1.5)) suggesting that binding of SO to cysteine residues of blood proteins is greatly preferred at the α position rather than the β position both in vitro and in vivo. The lowest detectable adduct concentration, using 10 mg of protein, was 8 pmol/g protein for SG,5 pmol/g protein for 1 PE, and 0.6 pmol/g protein for 2 PE. No significant change of adduct level was found during storage of proteins at -80℃ for 1 year.

Evaluation of HemogloBind^TMtreatment for preparation of samples for cholinesterase analysis

Acetylcholine is an essential neurotransmitter found throughout the nervous system. Its action on postsynaptic receptors is regulated through hydrolysis by various carboxylesterases, especially cholinesterases (ChEs). The acute toxicity of organophosphate (OP) compounds is directly linked to their action as inhibitors of ChE. One widely used assay for evaluating ChE activity is a spectrophotometric method developed by Ellman et al. When the enzyme source is from tissues or, in particular, blood, hemoglobin displays a spectrophotometric peak at the same wave-length used to analyze cholinergic activity. This creates a substantial background that interferes with the Ellman’s assay and must be overcome in order to accurately monitor cholinesterase activity. Herein, we directly compare blood processing methods: classical method (1.67 ± 0.30 U/mL) and HemogloBindTM treatment (1.51 ± 0.17 U/mL), and clearly demonstrate that pretreatment of blood samples with HemoglobindTM is both a sufficient and rapid sample preparation method for the assessment of ChE activity using the Ellman’s method.