Objective: To evaluate the value of next-generation sequencing (NGS) in the prevention and management of thalassemia. Methods: A systematic search was performed in eight databases including China Biomedical Literature Database, Chinese National Knowledge Infrastructure, Chinese Scientific Journals Database, Wanfang database, PubMed, EMBASE, Web of Science, and Cochrane Library from the inception to 1 June 2022. Stata 17.0 and Review Manager 5.4 were used for the meta-analysis. Results: Nine studies containing 14794 participants were included in the meta-analysis. Compared with the routine genetic testing (including Gap-PCR and reverse dot blot), NGS had higher detection rates in screening thalassemia (RR 1.22, 95% CI 1.13-1.31, P<0.01), particularly for the α-thalassaemia mutation carriers (RR 1.24, 95% CI 1.07-1.44, P<0.01). However, no significant difference was found in the screening of β-thalassemia (RR 1.10, 95% CI 0.99-1.23, P>0.05). Conclusions: Compared with routine genetic testing, NGS had a higher detection rate in general, particularly in the detection of α-thalassemia.

Objective: To assess the effectiveness and adverse drug reactions of all-oral regimens for patients with multidrug-resistant tuberculosis. Methods: This retrospective study was conducted at 10 Programmatic Management of Drug Resistant Tuberculosis sites in Punjab province of Pakistan. Patients receiving treatment for drug resistant tuberculosis from July 2019 to December 2020 with at least interim result i.e. 6th month culture conversion or final outcomes (cured, complete, lost to follow-up, failure, death) available, were included in the study. Data was extracted from electronic data management system. For the reporting and management of adverse drug events, active tuberculosis drug safety monitoring and management was implemented across all sites. All the data was analyzed using SPSS version 22. Results: Out of 947 drug resistant tuberculosis patients included in this study, 579 (68%) of the patients had final outcomes available. Of these, 384 (67.9%) successfully completed their treatment. Out of 368 (32%) patients who had their interim results available, all had their 6th month culture negative. Combining new medications was thought to result in serious adverse outcomes such as QT prolongation. However, this study did not record any severe adverse events among patients. Conclusions: All-oral regimens formulation guided by overall treatment effectiveness resulted in treatment outcomes comparable to those obtained with traditional injectable treatment.

Objective: To evaluate coagulation abnormalities and their relationship with bleeding manifestations among patients with dengue. Methods: This observational study was conducted on 292 adult dengue patients who were admitted to a tertiary care hospital of Western India from July 2021 to June 2022. Coagulation tests including prothrombin time (PT), international normalized ratio (INR), activated partial thromboplastin time (aPTT), fibrinogen, and D-dimer were performed. Patients were monitored for bleeding manifestations. Results: Coagulation abnormalities were reported in 42.8% of the patients. Overall, prolonged aPTT was the most common coagulation abnormality (40.8%), followed by low fibrinogen (38.7%), raised D-dimer (31.2%), raised INR (26.0%) and prolonged PT (19.2%). Bleeding manifestations were present in 19.9% patients. PT, INR, aPTT and D-dimer levels were significantly higher (P<0.01) and fibrinogen level was significantly lower (P<0.001) in patients with bleeding compared to patients without bleeding. Patients with bleeding had a significantly higher rate of all coagulation abnormalities than patients without bleeding (P<0.01). Conclusions: Patients with bleeding showed a significantly higher frequency of coagulation abnormalities compared to patients without bleeding. Patients with dengue should be assessed for coagulation abnormalities.

Objective: To determine the genetic diversity, natural selection and mutations in Plasmodium (P.) knowlesi drug resistant molecular markers Kelch 13 and dhps gene in clinical samples of Malaysia. Methods: P. knowlesi full-length gene sequences Kelch 13 gene (PkK13) from 40 samples and dhps gene from 30 samples originating from Malaysian Borneo were retrieved from public databases. Genetic diversity, natural selection, and phylogenetic analysis of gene sequences were analysed using DNAsp v5.10 and MEGA v5.2. Results: Seventy-two single nucleotide polymorphic sites (SNPs) across the full-length PkK13 gene (63 synonymous substitutions and 9 non-synonymous substitutions) with nucleotide diversity of π~0.005 was observed. Analysis of the full-length Pkdhps gene revealed 73 SNPs and π~0.006 (44 synonymous substitutions and 29 non-synonymous substitutions). A high number of haplotypes (PkK13; H=37 and Pkdhps; H=29) with haplotype diversity of Hd ~0.99 were found in both genes, indicating population expansion. Nine mutant alleles were identified in PkK13 amino acid alignment of which, 7 (Asp 3 Glu, Lys 50 Gln, Lys 53 Glu, Ser 123 Thr, Ser 127 Pro, Ser 149 Thr and Ala 169 Thr) were within the Plasmodium specific domain, 2 (VaI 372 Ile and Lys 424 Asn) were in the BTB/POZ domain and no mutation was observed within the kelch propeller domain. The 29 non-synonymous mutations in the Pkdhps gene were novel and only presented in exon 1 and 2. Conclusions: Monitoring the mutations from clinical samples collected from all states of Malaysia along with clinical efficacy studies will be necessary to determine the drug resistance in P. knowlesi.

Objective: To characterise a collection of pili-carrying and none pili-carrying pneumococcal isolates of clinical origin for serotypes, antibiotic resistance and genotype. Methods: In total, 42 clinical isolates were collected between October 2017 and December 2019. Those isolates were analysed for antimicrobial susceptibility, serotype distribution, detection of pneumococcal virulence and pilus genes. Multilocus sequence typing was performed only for piliated isolates, followed by phylogenetic analysis. Results: The common isolation sites among the pneumococcal isolates were tracheal aspirate (28.6%), blood (26.2%), and sputum (23.8%). Fifty percent isolates were resistant to erythromycin, tetracycline (50.0%) and trimethoprim-sulfamethoxazole (43.0%). The most frequent were serotypes 19F (28.6%), 6A/B (23.8%) and 19A (14.3%). Piliated isolates were detected in a small proportion (33.3%); 64.3% were multidrug-resistant. ST320 was the prevalent sequence type among the piliated isolates and genetically related to the Pneumococcal Molecular Epidemiology Network clones Taiwan 19F -14 (CC271). In the phylogenetic analysis, some piliated isolates showed a close association having similar ST320, carrying serotype 19A and both pilus genes indicating their clonal spread. Conclusions: Pneumococcal lineages of piliated isolates have been globally disseminated and pili could have played a role in the spread of antibiotic resistant clones.

Rationale: Neurocysticercosis is a neglected tropical disease caused by the larvae of Taenia solium and may occur in immunocompetent people. We report a case of diffuse parenchymal neurocysticercosis mimicking tuberculous meningitis in an immunocompetent patient. Patient concerns: A 24-year-old immunocompetent woman presented with serial focal to generalized seizure, left body weakness, left abducens palsy, chronic cough, and body weight loss. She came from an area where she was unlikely to have contact with Taenia solium. Diagnosis: Diffuse parenchymal neurocysticercosis. Intervention: Valproic acid, dexamethasone and 15 months of albendazole therapy. Outcome: Complete recovery without any neurological sequelae. Lessons: Prolonged anthelminthic treatment may benefit neurocysticercosis patients with multiple cerebral parenchymal lesions.

Objective: To determine the suitable ecological habitats of Aedes (Ae.) aegypti and Ae. albopictus in Iran due to climate change by the 2070s. Methods: All data relating to the spatial distribution of Ae. aegypti and Ae. albopictus worldwide, which indicated the geographical coordinates of the collection sites of these mosquitoes, were extracted from online scientific websites and entered into an Excel file. The effect of climatic and environmental variables on these mosquitoes was evaluated using the MaxEnt model in the current and future climatic conditions in the 2030s, 2050s, and 2070s. Results: The most suitable areas for the establishment of Ae. aegypti are located in the southern and northern coastal areas of Iran, based on the model outputs. The modelling result for suitable ecological niches of Ae. albopictus shows that in the current climatic conditions, the southern half of Iran from east to west, and parts of the northern coasts are prone to the presence of this species. In the future, some regions, such as Gilan and Golestan provinces, will have more potential to exist/establish Ae. albopictus. Also, according to the different climate change scenarios, suitable habitats for this species will gradually change to the northwest and west of the country. The temperature of the wettest season of the year (Bio8) and average annual temperature (Bio1) were the most effective factors in predicting the model for Ae. aegypti and Ae. albopictus, respectively. Conclusions: It is required to focus on entomological studies using different collection methods in the vulnerable areas of Iran. The future modelling results can also be used for long-term planning to prevent the entry and establishment of these invasive Aedes vectors in the country.

Objective: To assess the knowledge of healthcare workers on the measles vaccine and its cold chain management. Method: An institutional-based cross-sectional study was conducted from February 1 to March 30, 2022 in Gondar City Administration public health institutions among 165 healthcare workers. Data were collected using a structured questionnaire. In addition, an on-spot observation checklist was used to assess the availability, status and management of the cold chain. A logistic regression model was used to assess the relationship between the outcome and predictor variables. Crude and adjusted odds ratios were calculated with 95% confidence intervals. Results: Overall, 87 (52.7%; 95% Cl 44.8%-60.5%) of the healthcare workers had unsatisfactory knowledge regarding the measles vaccine and its cold chain management. One hundred thirty-six (82.4%) healthcare workers correctly mentioned the recommended range of temperature (2-8 °C) for measles vaccine storage. Healthcare workers aged 18-29 years (P=0.001) and 30-44 years (P=0.014) were observed as determinants of unsatisfactory knowledge on the measles vaccine and its cold chain management. One hundred and five (63.6%) of the healthcare workers did not correctly mention the type of measles vaccine used in routine immunization. More than one-third (36.4%) of the healthcare workers perceived that the measles vaccine is not safe and could cause measles. Conclusions: More than half of the healthcare workers in the study area had unsatisfactory knowledge on the measles vaccine and its cold chain management. It is necessary to provide technical support and in-service training for healthcare workers to ensure optimal immunization effectiveness.

Objective: To analyze data on socio-demographic and clinical characteristics of SARS-CoV-2 infected population whose samples were received from Medical Research Institute, Sri Lanka. Methods: Laboratory based retrospective study was done on patient samples which were tested positive for SARS-CoV-2 by National Reference Virology Laboratory at the Medical Research Institute, Sri Lanka, from November, 2020 to November, 2021. Data on socio-demographic characteristics and clinical presentation of 13 126 patients were examined. Results: The mean age of the study population was (36.0±7.2) years and the majority were men (64.0%). The highest number of positive cases were found in the 21-30 years-of-age group. Two distinct peaks were noted in the incidence of SARS-CoV-2 positive individuals. In addition, 42.5% of the positive samples tested positive (42.5%) were from Medical Officer of Health collection centres. Furthermore, 60.6% (7 951) of the infected subjects were asymptomatic whereas the remaining were symptomatic. The highest percentage of symptomatic patients were observed in the 91-100 years-of-age group while the highest asymptomatic subjects were found in the 31-40 years-of-age group. The percentage of asymptomatic children (65.3%) was significantly (P<0.05) higher than that of adults (43.4%). Conclusions: The findings of this study aid decision makers to focus on the vulnerable groups, and geographic and temporal distribution of patients in the public health strategies that aim at preventing the spread of the disease and reducinig its mortalities.

Objective: To present platelet large cell ratio (P-LCR), reticulocyte, and immature reticulocyte fraction (IRF) values as novel parameters in diagnosis and response to treatment in patients developing sacroiliitis. Methods: Sixty-eight patients with clinical symptoms and Brucella standard tube agglutination (Wright) or Brucella Coombs agglutination test titers ≥1:160 were included in the study. Two groups were established, one developing sacroiliitis and another with no sacroiliitis development. P-LCR, reticulocyte, and IRF levels were measured using a Sysmex XN-9000 device (Japan). These were then compared between the two groups. Results: Reticulocyte (P=0.037) and IRF (P=0.026) levels were significantly lower among the patients developing sacroiliitis compared to the non-sacroiliitis group, while P-LCR (P=0.003) levels were significantly higher. P-LCR had the most powerful correlation with sacroiliitis development. Significant negative correlation was observed between reticulocyte, IRF levels and sacroiliitis. Conclusions: Elevated P-LCR levels were observed as a marker of persisting inflammation in patients developing sacroiliitis, while low reticulocyte and IRF levels secondary to bone marrow involvement were detected. These three parameters emerged as highly significant markers in terms of diagnosis and reflecting responses to treatment in organ involvement such as sacroiliitis in brucellosis. These are presented as inexpensive, and easily accessible novel parameters.