Exosomal hsa_circ_000200 as a potential biomarker and metastasis enhancer of gastric cancer via miR-4659a/b-3p/HBEGF axis.

BACKGROUND: Exosome, a component of liquid biopsy, loaded protein, DNA, RNA and lipid gradually emerges as biomarker in tumors. However, exosomal circRNAs as biomarker and function mechanism in gastric cancer (GC) are not well understood. METHODS: Differentially expressed circRNAs in GC and healthy people were screened by database. The identification of hsa_circ_000200 was verified by RNase R and sequencing, and the expression of hsa_circ_000200 was evaluated using qRT-PCR. The biological function of hsa_circ_000200 in GC was verified in vitro. Western blot, RIP, RNA fluorescence in situ hybridization, and double luciferase assay were utilized to explore the potential mechanism of hsa_circ_000200. RESULTS: Hsa_circ_000200 up-regulated in GC tissue, serum and serum exosomes. Hsa_circ_000200 in serum exosomes showed better diagnostic ability than that of tissues and serum. Combined with clinicopathological parameters, its level was related to invasion depth, TNM staging, and distal metastasis. Functionally, knockdown of hsa_circ_000200 inhibited GC cells proliferation, migration and invasion in vitro, while its overexpression played the opposite role. Importantly, exosomes with up-regulated hsa_circ_000200 promoted the proliferation and migration of co-cultured GC cells. Mechanistically, hsa_circ_000200 acted as a "ceRNA" for miR-4659a/b-3p to increase HBEGF and TGF-ß/Smad expression, then promoted the development of GC. CONCLUSIONS: Our findings suggest that hsa_circ_000200 promotes the progression of GC through hsa_circ_000200/miR-4659a/b-3p/HBEGF axis and affecting the expression of TGF-ß/Smad. Serum exosomal hsa_circ_000200 may serve as a potential biomarker for GC.

Mendelian randomization studies of depression: evidence, opportunities, and challenges.

BACKGROUND: Major depressive disorder (MDD) poses a significant social and economic burden worldwide. Identifying exposures, risk factors, and biological mechanisms that are causally connected to MDD can help build a scientific basis for disease prevention and development of novel therapeutic approaches. METHODS: In this systematic review, we assessed the evidence for causal relationships between putative causal risk factors and MDD from Mendelian randomization (MR) studies, following PRISMA. We assessed methodological quality based on key elements of the MR design: use of a full instrumental variable analysis and validation of the three key MR assumptions. RESULTS: We included methodological details and results from 52 articles. A causal link between lifestyle, metabolic, inflammatory biomarkers, particular pathological states and MDD is supported by MR investigations, although results for each category varied substantially. CONCLUSIONS: While this review shows how MR can offer useful information for examining prospective treatment targets and better understanding the pathophysiology of MDD, some methodological flaws in the existing literature limit reliability of results and probably underlie their heterogeneity. We highlight perspectives and recommendations for future works on MR in psychiatry.

Research Progress of Forensic Diagnosis Approaches of Early Acute Myocardial Infarction.

The postmortem diagnosis of acute myocardial infarction (AMI), especially the postmortem diagnosis of early AMI that died immediately after onset or within 1 hour, has always been a difficulty in forensic identification. This article reviews the forensic application of diagnosis and analysis methods for AMI postmortem diagnosis including autopsy imaging, histomorphology, immunohisto-chemistry, biochemical marker and molecular biology diagnosis, and explores the feasible scheme of early postmortem diagnosis in AMI.

Change Rule of Diatom in Zhangweixin River in Dezhou.

OBJECTIVES: To study the annual variation of diatoms in Zhangweixin River, to provide theoretical support by using diatom examination to estimate the time and place of the corpse entering water, and to establish a diatom database. METHODS: Samples were taken from 4 sampling sites in Decheng section of Zhangweixin River for 12 consecutive months. Non-metric multi-dimensional scaling (NMDS) analysis was performed on the species and content of diatom samples. Based on the sampling site of Tianqu Road, Sprensen similarity coefficient analysis was conducted with the data of other 3 sites in Decheng section and Leling section of Zhangweixin River and Ningjin section in previous studies. RESULTS: The number of diatom species was positively correlated with diatom content. The average diatom content in different months ranged from 1 054 to 13 041/10 mL, and the species composition ranged from 8 to 16, with differences in dominant species. The similarity coefficient of diatom species within 2 km were all higher than 0.956 52, which could not be distinguished effectively. The similarity coefficients of Leling section and Ningjin section were 0.736 84 and 0.588 24 respectively, which could be effectively distinguished. CONCLUSIONS: The species and content of diatom vary in different months in Zhangweixin River, and the composition of diatom species is different in different basins, which can provide reference for estimating the time and place of the corpse entering water in the river.

Metal trabecular bone reconstruction system better improves clinical efficacy and biomechanical repair of osteonecrosis of the femoral head than free vascularized fibular graft: A case-control study.

In this study, we aim to compare and analyze the biomechanical repair and clinical efficacy of osteonecrosis of the femoral head (ONFH) with the use of metal trabecular bone reconstruction system and free vascularized fibular graft. The study enrolled 66 adult patients from medical records of nontraumatic ARCO 2A-3B stage ONFH. A simple ONFH model without surgical treatment was established in 13 cases, 29 cases were treated with metal trabecular bone reconstruction system, and 24 cases were treated with free vascularized fibular graft. Computer-recognized and extracted femur outlines were imported, and three-dimensional reconstructions were performed. The stress concentration and stress peak value were analyzed, and the Harris score, visual analog scale pain score, and operation status of the above patients were compared. Finally, quality of life assessment was performed using SF-36 scale. Metal trabecular bone reconstruction system provided less operation time, blood loss, and the total length of postoperative hospital stay than free vascularized fibular graft. Metal trabecular bone reconstruction system promoted bone reconstruction, increased bone mineral density and Harris score. The total clinical effective rate of young patients (20-40 years) was higher than that of older patients (41-60 years). Metal trabecular bone reconstruction system provided higher physical component summary, mental component summary, and role/social component summary than free vascularized fibular graft. This study demonstrates that both metal trabecular bone reconstruction system and free vascularized fibular graft can prevent or delay the progression of ONFH, while metal trabecular bone reconstruction system is a better choice because of better short-term clinical efficacy.

The Efficacy of Imrecoxib and Celecoxib in Axial Spondyloarthritis and Their Influence on Serum Dickopff-Related Protein 1 (DKK-1) Levels.

BACKGROUND To observe and demonstrate therapeutic effects and side effects of two selective COX-2 inhibitors, imrecoxib and celecoxib, on patients with axial spondyloarthritis (axSpA) and observe the correlation between imaging scores and serum DKK-1 levels. MATERIAL AND METHODS Sixty patients with axSpA were randomly assigned to receive 200 mg imrecoxib or 200 mg celecoxib twice daily. Fifty-one patients who completed follow-up were included in the study. At baseline, week 4, and week 12, the clinical parameters, inflammatory markers (ESR, CRP), and adverse reactions were recorded. Serum DKK-1 levels were investigated by enzyme-linked immunosorbent assay. Radiographic scores were calculated by sacroiliac joint SPARCC (Spondyloarthritis Research Consortium of Canada) score method at baseline serum DKK-1 levels and week 12. RESULTS Patients in the imrecoxib group (n=25) and patients in the celecoxib group (n=26) were improved at week 4. At week 12, all clinical parameters and inflammatory markers were improved in the two groups and the differences was not statistically significant. Serum DKK-1 levels were decreased and the differences were not statistically significant. Serum DKK-1 levels in patients in the imrecoxib group at baseline were negatively correlated with all study parameters, while those in the celecoxib group had correlations with BASFI (r=-0.048, p=0.027) and Schober test (r=0.437, p=0.048), without any correlation with other clinical parameters or inflammatory markers. CONCLUSIONS Patients experienced significant improvement in disease activity, functional parameters, and inflammatory markers when treated with selective COX-2 inhibitors for 12 weeks, and the efficacy of imrecoxib was not inferior to celecoxib. Selective COX-2 inhibitors imrecoxib and celecoxib had no obvious effects on serum DKK-1 levels.

Multiple cannulated screws vs. dynamic hip screws for femoral neck fractures : A meta-analysis.

OBJECTIVE: Various fixation devices have been reported for stabilization of femoral neck fractures. Numerous studies on arthroplasty versus internal fixation devices in the treatment of femoral neck fractures have been performed, but the optimal approach for internal fixation has not been analyzed. METHODS: A meta-analysis and system evaluation were performed to compare clinical effects. We searched PubMed, Embase, Cochrane Library, and Web of ScienceTM for randomized, controlled trials (RCTs) comparing multiple cannulated screws (MCS) with dynamic hip screws (DHS) and analyzed the failure rate of operations, the reoperation rate, and postoperative complications. Risk ratios (RRs) and mean differences from each trial were pooled using random effects or fixed effects models, depending on study heterogeneity. The analysis was performed using RevMan5.2. RESULTS: In this meta-analysis, 592 femoral neck fractures from 7 studies were assessed, and the meta-analysis results indicated significant differences in reoperation (RR 1.44, 95% confidence interval [CI] 1.10-1.88, P = 0.008) and failure rate (RR 2.28, 95% CI 1.10-4.72, P = 0.03), but no significant differences in the rate of postoperative complications between the MCS group and DHS group. CONCLUSIONS: DHS fixation has a larger skin incision and more soft tissue dissection, but it is associated with lower rates of fixation failure, reoperation, and overall rate of postoperative complications, and its use in elderly patients with osteoporosis is still recommended due to simplicity, efficacy, and high overall success rate. Multicenter RCTs with large samples are needed to better understand the comparative efficacy and safety of MCS and DHS in femoral neck fractures of restricted fracture type.

Cytotoxic taccalonolides and withanolides from Tacca chantrieri.

Six new taccalonolides, taccalonolides AT-AY (1-6), and two new withanolides, chantriolides D and E (7 and 8), together with ten known compounds (9-18), have been isolated from whole plants of Tacca chantrieri. The structures, including the absolute configurations of some of the compounds, were determined by spectroscopic and chemical methods. All compounds were evaluated for their in vitro cytotoxicity against five tumor cell lines. Compounds 9, 10, 13-15, and 17 exhibited cytotoxic activity, with IC50 values of 1.13-5.71 µM, while compound 7 showed selective cytotoxicity. The results indicated that taccalonolides with a six-membered lactone moiety located at C-15 and C-24 were devoid of cytotoxicity against five tumor cell lines (> 10 µM).

[HPLC enantioseparation, absolute configuration determination and anti-HIV-1 activity of (±)-F18 enantiomers].

Racemic (±)-F18 (10-chloromethyl-11-demethyl-12-oxo-calanolide A), an analog of nature product (+)-calanolide A, is a new anti-HIV-1 nonnucleoside reverse transcript inhibitor (NNRTI). A successful enantioseparation of (±)-F18 offering (R)-F18 and (S)-F18 was achieved by a chiral stationary phase prepared HPLC. Their absolute configurations were determined by measurement of their electronic circular dichroisms combined with modem quantum-chemical calculations. Further investigation revealed that (R)-F18 and (S)-F18 shared a similar anti-HIV activities, however, (R)-F18 was more potent than (S)-F18 against wild-type virus, K101E mutation and P225H mutation pseudoviruses.

[Reflection on Medical Treatment of Multi-drug Resistance Tuberculosis: The Necessity of Chinese Medicine Holistic View].

Causative factors of multi-drug resistance tuberculosis (MDR-TB) were analyzed from iatrogenic angles, patients themselves, and society. Reviewed was the development of treatment strategies for MDR-TB from directly observed treatment short-course (DOTS) to DOTS-Plus. The history of Chinese medicine (CM) fighting TB and characteristics at the present stage were also analyzed. Authors pointed out that CM pays attention not only to killing pathogens and confirms the necessity of getting rid of pathogens, but also to cascade response caused by pathogens. It also regards the occurrence and development of MDR-TB as a whole by combining patients' conditions, climatic, geographic, psychological, and social factors. Authors believed that therapeutic principles under guidance of CM holistic view are of positive significance and inspiration in treating MDR-TB, and emphasized holistic view as basic strategies for treating MDR-TB, but not a single countermeasure.

[A method for assessing the total viable count of fresh meat based on hyperspectral scattering technique].

The objective of this study is to develop a hyperspectral imaging system to predict the bacteria total viable count in fresh pork. The hyperspectral scattering data were curvefitted by different fitting methods, and correlation differences of models were compared based on the bacteria total viable count of fresh pork, thus providing modeling basis of device for future study. Total 63 fresh pork samples which was used in the experiment were stored at 4 degrees C in the refrigerator of constant temperature. Experiment was performed everyday for 15 days. 4 or 5 random samples were used each day for the experiment. Hyperspectral scattering images and spectral scattering optical data in the wavelength region of 400 to 1 100 nm were acquired from the surface of all of the pork samples. Lorentz and Gompertz function and the modified function was applied to fit the scattering profiles of pork samples. Different parameters could be obtained by Lorentz and Gompertz fitting and the modified function fitting. The different parameters could represent the optical characteristic of the scattering profiles. The standard values of the bacteria total viable count of pork were obtained by classical microbiological plating methods. Because the standard value of the bacteria total viable count was big, log10 of the bacteria total viable count obtained by classical microbiological plating was used to simplify the calculation. Both individual parameters and integrated parameters were explored to develop the models. The multi-linear regression statistical approach was used to establish the models for predicting pork the bacteria total viable count. Both Lorentz and Gompertz function and the modified function included three and four parameters formula. The results showed that correlation coefficient of the models is higher with Lorentz three parameters combination, Lorentz four parameters combination and Gompertz four parameters combination than the individual parameters and other two or three integrated parameters. The three models' correction set and prediction set correlation coefficients were 0.93, 0.96, 0.96 and 0.90, 0.90, 0.92, and the corresponding standard deviation were 0.47, 0.44, 0.39 and 0.56, 0.46, 0.42. Correlation was best with Gompertz four parameters. The device system will select the best correlation and the best stability of the model as the final model.

CircSCAF8 promotes growth and metastasis of prostate cancer through the circSCAF8-miR-140-3p/miR-335-LIF pathway.

Circular RNAs (circRNAs) have been increasingly linked to cancer progression. However, the detailed biological functions of circRNAs in prostate cancer (PCa) remain unclear. Using high-throughput circRNA sequencing, we previously identified 18 urine extracellular vesicle circRNAs that were increased in patients with PCa compared with those with benign prostatic hyperplasia. Spearman correlation analysis of the expression levels of the 18 circRNAs between the tumor tissue and matched urine extracellular vesicles in 30 PCa patients showed that circSCAF8 had the highest R2 (R2 = 0.635, P < 0.001). The Cox proportional hazards regression model was used to estimate the effect of circSCAF8 on progression-free survival. The in vitro and in vivo functional experiments were implemented to investigate the effects of circSCAF8 on the phenotype of PCa. We found that the knockdown of circSCAF8 in PCa cells suppressed the proliferation, migration, and invasion ability, while overexpression of circSCAF8 had the opposite effects. Similar results were observed in vivo. In a cohort of 85 patients who had undergone radical prostatectomy, circSCAF8 expression in PCa tissues was a powerful predictor of progression-free survival (HR = 2.14, P = 0.022). Mechanistically, circSCAF8 can function by binding to both miR-140-3p and miR-335 to regulate LIF expression and activate the LIF-STAT3 pathway that leads to the growth and metastasis of PCa. Collectively, our findings demonstrate that circSCAF8 contributes to PCa progression through the circSCAF8-miR-140-3p/miR-335-LIF pathway.

[Acetylsalicylic acid strengthens the effects of ANISpm against hepatocellular carcinoma and its molecular mechanism].

The objective of this study is to examine the effects of ANISpm, a novel polyamine naphthalimide conjugate, with acetylsalicylic acid against hepatocellular carcinoma in vivo and in vitro and elucidate its potential molecular mechanism. The proliferation inhibition was detected by MTT assay. Cell apoptosis, intracellular fluorescence intensity and mitochondrial membrane potential (MMP) were detected by high content screening (HCS) analysis. Polyamines content was analyzed by reverse-phase high performance liquid chromatography Protein expression levels were quantified by Western blotting assay. The combination treatment strongly inhibited cell proliferation, induced cell apoptosis in HepG2 cells and H22 hepatoma cells, which was mediated by enhanced ANISpm uptake via up-regulation of spermidine/spermine N1-acetyltransferase (SSAT) and depression of intracellular polyamine. Furthermore, this synergistic apoptosis was involved in mitochondria and death-receptor signal pathway. All these findings demonstrated that the combination treatment with acetylsalicylic acid and ANISpm resulted in synergistic antitumor effects on hepatoma cells. Thus, combination therapy with these agents may be useful as a potential template for the development of better chemotherapeutic strategy against hepatoma.

[Inhibition of culture supernatant of mesenchymal stem cells on macrophages RAW264.7 activated by soluble egg antigen of Schistosoma japonicum].

OBJECTIVE: To observe the inhibitive effect of rat mesenchymal stem cells (MSC) culture supernatant on macrophages activated by soluble egg antigen (SEA) of Schistosoma japonicum. METHODS: To select optimal SEA effecting concentration and time, macrophages RAW264.7 were induced by 5, 10, 20 or 40 microg/ml SEA for 12 h, or by 20 microg/ml SEA for 4, 8, 12 or 24 h before examination of TNF-alpha mRNA by RT-PCR. Macrophages were divided into five groups, i.e. negative control group, SEA group, SEA+MSC supernatant group (MSC group), SEA+NRK-52E supernatant group (NRK-52E group) and SEA+DMEM group (DMEM group). Except negative control group, macrophages in other four groups were induced by 20 microg/ml SEA for 12 h. SEA was then removed from MSC group, NRK-52E group and DMEM group and replaced with MSC supernatant, NRK-52E supernatant and DMEM, respectively. Morphology of macrophages in each group was observed by microscope after cultured with supernatant for 12 h. TNF-alpha mRNA in macrophages was detected by real-time quantitative PCR after cultured with supernatant for 12 h and 24 h. TGF-beta1 in macrophages was observed by Western blotting analysis after cultured with supernatant for 12 h. Macrophage proliferation was tested by MTT method after cultured with supernatant for 24 h and 48 h. RESULTS: The optimal SEA concentration and time for macrophage activation was 20 microg/ml and 12 h, respectively. Compared with SEA group, NRK-52E group, and DMEM group, macrophages in MSC group were round and small with less pseudopodia after cultured with supernatant for 12 h. TNF-alpha mRNA after cultured with MSC supernatant for 12 h and 24 h was (1.0 +/- 0.4) and (1.0 +/- 0.5) fold of negative control group, respectively, significantly less than NRK-52E group [(10.4 +/- 3.9) and (16.5 +/- 5.0) fold] (12 h: P < 0.05; 24 h: P < 0.01) and DMEM group [(6.0 +/- 2.1) and (2.4 +/- 0.7) fold] (P < 0.05). The grey density image analysis of TGF-beta1/GAPDH was 0.31 +/- 0.10 in MSC group, much lower than 0.88 +/- 0.10 in NRK-52E group (P < 0.01) and 0.58 +/- 0.06 in DMEM group (P < 0.05) after cultured with supernatant for 12 h. After 48 h culture, A490 of macrophages in MSC group was 0.22 +/- 0.05, much lower than 0.53 +/- 0.02 in NRK-52E group and 0.31 +/- 0.03 in DMEM group (P < 0.05). CONCLUSION: MSC supernatant can inhibit activation and proliferation of macrophages which were induced by SEA of S. japonicum.

Prediction of potential prognostic biomarkers in metastatic prostate cancer based on a circular RNA-mediated competing endogenous RNA regulatory network.

Recently, studies on competing endogenous RNA (ceRNA) networks have become prevalent, and circular RNAs (circRNAs) have crucial implications for the development and progression of carcinoma. However, studies relevant to metastatic prostate cancer (mPCa) are scant. This study aims to discover potential ceRNAs that may be related to the prognosis of mPCa. RNA-Seq data were obtained from the MiOncoCirc database and Gene Expression Omnibus (GEO). Differential expression patterns of RNAs were examined using R packages. Circular RNA Interactome, miRTarBase, miRDB and TargetScan were applied to predict the corresponding relation between circRNAs, miRNAs and mRNAs. The Gene Ontology (GO) annotations were performed to present related GO terms, and Gene Set Enrichment Analysis (GSEA) tools were applied for pathway annotations. Moreover, survival analysis was conducted for the hub genes. We found 820 circRNAs, 81 miRNAs and 179 mRNAs that were distinguishingly expressed between primary prostate cancer (PCa) and mPCa samples. A ceRNA network including 45 circRNAs, 24 miRNAs and 56 mRNAs was constructed. In addition, the protein-protein interaction (PPI) network was built, and 10 hub genes were selected by using the CytoHubba application. Among the 10 hub genes, survival analysis showed that ITGA1, LMOD1, MYH11, MYLK, SORBS1 and TGFBR3 were significantly connected with disease-free survival (DFS). The circRNA-mediated ceRNA network provides potential prognostic biomarkers for metastatic prostate cancer.

[Electronic focusing high-nergy extracorporeal shockwave combined with conventional oral medicine for the treatment of ankylosing spondylitis].

OBJECTIVE: To observe the progress of disease activity and sacroiliac joint imaging in patients with ankylosing spondylitis treated by extracorporeal shockwave combined with conventional oral medicine, and find a new safe and effective therapeutic method. METHODS: The clinical data of 30 patients with ankylosing spondylitis treated from January 2018 to December 2018 were retrospectively analyzed. Including 20 males and 10 females, aged from 18 to 50 years with an average of (34.50±9.60) years. All 30 patients had different degrees of sacroiliac joint bone marrow edema on MRI before treatment. Thirty patients were divided into treatment group and control group according to different treatment methods. Among them, 15 cases in control group were treated with non-steroidal anti-inflammatory drugs and sulfasalazine enteric-coated tablets, for the 15 cases in treatment group, in addition to oral medicine in line with control group, electronic focusing high-energy extracorporeal shockwave therapy was added. The course of disease, age, pre- and post-treatment erythrocyte sedimentation rate, C-reactive protein in the two groups were analyzed; and visual analogue scale (VAS) and spondyloarthritis research consortium Canada (SPARCC) scoring system were used to evaluate the pain of the sacroiliac joint and the structural damage of the sacroiliac joint;Bath ankylosing spondylitis disease activity index (BASDAI) was calculated. RESULTS: All patients were followed up for at least 3 months. One month after treatment, VAS, and SPARCC scores in treatment group were significantly better than in control group (P<0.05). After 1 month of treatment, there was no significant difference in BASDAI, erythrocyte sedimentation rate and C-reactive protein between two groups(P>0.05). VAS, BASDAI, SPARCC, erythrocyte sedimentation rate, and C-reactive protein of all patients after treatment were significantly improved compared with those before treatment (P< 0.01). CONCLUSION: Electronic focusing high-energy extracorporeal shockwave combined with conventional oral medicine in the treatment of ankylosing spondylitis has a good clinical effect in rapidly relieving pain, improving disease activity, and preventing imaging progress. In addition, it is safe and non-invasive, which is worthy of clinical application.

The nucleolus functions as the compartment for histone H2B protein degradation.

Histones are main components of chromatin, and the protein levels of histones significantly affect chromatin assembly. However, how histone protein levels are regulated, especially whether and how histones are degraded, is largely unclear. Here, we found that histone H2B is mainly degraded through the proteasome-mediated pathway, and the lysine-120 site of H2B is essential for its K48-linked polyubiquitination and degradation. Moreover, the degradation-impaired H2BK120R mutant shows an increased nucleolus localization, and inhibition of the proteasome results in an elevated nucleolus distribution of wild-type H2B, which is similar to that of H2BK120R mutants. More importantly, the nucleolus fractions can ubiquitinate and degrade the purified H2B in vitro, suggesting that the nucleolus, in addition to its canonical roles regulating ribosome genesis and protein translation, likely associates with H2B degradation. Therefore, these findings revealed a novel mechanism for the regulation of H2B degradation in which a nucleolus-associated proteasome pathway is involved.

[A rapid nondestructive measurement method for assessing the total plate count on chilled pork surface].

The present paper proposed a method based on the hyperspectral technology for rapidly, nondestructively quantify the total plate count on chilled pork surface. In the research, 50 chilled pork samples stored at 4 degrees C for 1-14 days were used to study the relationship between the total plate count on chilled pork surface and their hyperspectral images collected in 400-1 100 nm. Two models were established using MLR and PLSR methods, and the prediction showed that they can both give satisfactory results with R(v) = 0.886 and 0.863 respectively. The overall research demonstrates that the hyperspectral technology can well quantify the total plate count on chilled pork surface, and so indicates that it is a valid tool to assess the quality and safety properties of chilled pork in the future.

Attenuated P. falciparum Parasite Shows Cytokine Variations in Humanized Mice.

A recently developed humanized mouse has been used to assess the immune response evoked against the isolated attenuated C9 parasite clone (C9-M; carrying a single insertion disrupting the open reading frame (ORF) of PF3D7_1305500) of Plasmodium falciparum. Significant human RBC engraftment was achieved by ameliorating the residual non-adaptive immune response using clodronate-loaded liposome treatment. Controlled reactive professional phagocytic leukocytes in immunodeficient mice allowed for sizeable human blood chimerism and injected huRBCs acted as bona fide host cells for P. falciparum. huRBC-reconstituted immunodeficient mice received infectious challenge with attenuated P. falciparum C9 parasite mutants (C9-M), complemented (C9-C), and wild type (NF54) progenitors to study the role of immune effectors in the clearance of the parasite from mouse circulation. C9-M and NF54 parasites grew and developed in the huRBC-reconstituted humanized NSG mice. Further, the presence of mutant parasites in deep-seated tissues suggests the escape of parasites from the host's immune responses and thus extended the survival of the parasite. Our results suggest an evasion mechanism that may have been employed by the parasite to survive the mouse's residual non-adaptive immune responses. Collectively, our data suggest that huRBCs reconstituted NSG mice infected with attenuated P. falciparum is a valuable tool to explore the role of C9 mutation in the growth and survival of parasite mutants and their response to the host's immune responses. This mouse might help in identifying novel chemotherapeutic targets to develop new anti-malarial drugs.