Progress of mitochondrial and endoplasmic reticulum-associated signaling and its regulation of chronic liver disease by Chinese medicine.

The endoplasmic reticulum (ER) is connected to mitochondria through mitochondria-associated ER membranes (MAMs). MAMs provide a framework for crosstalk between the ER and mitochondria, playing a crucial role in regulating cellular calcium balance, lipid metabolism, and cell death. Dysregulation of MAMs is involved in the development of chronic liver disease (CLD). In CLD, changes in MAMs structure and function occur due to factors such as cellular stress, inflammation, and oxidative stress, leading to abnormal interactions between mitochondria and the ER, resulting in liver cell injury, fibrosis, and impaired liver function. Traditional Chinese medicine has shown some research progress in regulating MAMs signaling and treating CLD. This paper reviews the literature on the association between mitochondria and the ER, as well as the intervention of traditional Chinese medicine in regulating CLD.

Multimodal mechanisms of pathogenic variants in the signal peptide of FIX leading to hemophilia B.

ABSTRACT: Signal peptide (SP) is essential for protein secretion, and pathogenic variants in the SP of factor IX (FIX) have been identified in hemophilia B (HB). However, the underlying mechanism for the genotype-phenotype correlation of these variants has not been well studied. Here, we systematically examined the effects of 13 pathogenic point variants in the SP of FIX using different approaches. Our results showed that these point variants lead to HB by missense variants and/or aberrant premessenger RNA (pre-mRNA) splicing. The missense variants in a hydrophobic core (h-region) mainly affected the cotranslational translocation function of the SP, and those in C-terminal containing cleavage site (c-region) caused FIX deficiency mainly by disturbing the cotranslational translocation and/or cleavage of the SP. Almost absolute aberrant pre-mRNA splicing was only observed in variants of c.82T>G, but a slight change of splicing patterns was found in variants of c.53G>T, c.77C>A, c.82T>C, and c.83G>A, indicating that these variants might have different degrees of impact on pre-mRNA splicing. Although two 6-nt deletion aberrant pre-mRNA splicing products caused FIX deficiency by disturbing the SP cleavage, they could produce some functional mature FIX, and vitamin K could increase the secretion of functional FIX. Taken together, our data indicated that pathogenic variants in the SP of FIX caused HB through diverse molecular mechanisms or even a mixture of several mechanisms, and vitamin K availability could be partially attributed to varying bleeding tendencies in patients carrying the same variant in the SP.

Clinicopathological factors predict residual lymph node metastasis in locally advanced rectal cancer with ypT0-2 after neoadjuvant chemoradiotherapy.

PURPOSE: Residual lymph node metastases (RLNM) remained a great concern in the implementation of organ-preserving strategies and led to poor prognosis in locally advanced rectal cancer (LARC). In this study, we aimed to identify the clinicopathological factors correlated with RLNM in LARC patients with ypT0-2 after neoadjuvant chemoradiotherapy (NCRT). METHODS: We retrospectively analyzed 417 patients histologically diagnosed middle-low LARC after NCRT and total mesorectal excision (TME), whose pathological staging was ypT0-2. All patients received pelvic magnetic resonance imaging (MRI) before NCRT. The radiation doses were 50-50.6 Gy for the planning gross tumor volume and 41.8-45 Gy for the planning target volume, respectively. A nomogram for predicting RLNM was constructed using a binary logistic regression. Nomogram performance was assessed by receiver operating characteristic (ROC) curve, calibration curve, decision curve analysis (DCA) and clinical impact curve (CIC). RESULTS: After surgery, 191 patients (45.8%) were ypT0, 43 patients (10.3%) were ypT1 and 183 patients (43.9%) were ypT2, and a total of 49 patients (11.8%) were found the presence of RLNM. Multivariable analyses identified MRI-defined mesorectal fascia (MRF)-positive, high-grade histopathology at biopsy, advanced ypT-category, and the presence of perineural invasion (PNI) as the predictive factors. The nomogram, incorporating all these predictors, showed good discrimination and calibration efficacy, with the areas under the ROC curve of 0.690 (95% CI: 0.610-0.771). Both DCA and CIC demonstrated that this nomogram has good clinical usefulness. CONCLUSION: The nomogram model can predict RLNM in patients with ypT0-2 tumors. It can help select suitable patients for performing organ-preserving strategies after NCRT.

Research progress of the regulation of orphan nuclear receptors on chronic liver diseases / 生理学报

The development of chronic liver disease can be promoted by excessive fat accumulation, dysbiosis, viral infections and persistent inflammatory responses, which can lead to liver inflammation, fibrosis and carcinogenesis. An in-depth understanding of the etiology leading to chronic liver disease and the underlying mechanisms influencing its development can help identify potential therapeutic targets for targeted treatment. Orphan nuclear receptors (ONRs) are receptors that have no corresponding endogenous ligands to bind to them. The study of these ONRs and their biological properties has facilitated the development of synthetic ligands, which are important for investigating the effective targets for the treatment of a wide range of diseases. In recent years, it has been found that ONRs are essential for maintaining normal liver function and their dysfunction can affect a variety of liver diseases. ONRs can influence pathophysiological activities such as liver lipid metabolism, inflammatory response and cancer cell proliferation by regulating hormones/transcription factors and affecting the biological clock, oxidative stress, etc. This review focuses on the regulation of ONRs, mainly including retinoid related orphan nuclear receptors (RORs), pregnane X receptor (PXR), leukocyte cell derived chemotaxin 2 (LECT2), Nur77, and hepatocyte nuclear factor 4α (HNF4α), on the development of different types of chronic liver diseases in different ways, in order to provide useful references for the therapeutic strategies of chronic liver diseases based on the regulation of ONRs.

Study on mechanism of curcumol against liver fibrosis based on autophagy and apoptosis of hepatic stellate cells / 中国中药杂志

The present study clarified the molecular mechanism of curcumol against liver fibrosis based on its effects on the autopha-gy and apoptosis of hepatic stellate cells. The hepatic stellate cells were divided into a blank control group, a transforming growth factor-β1(TGF-β1)(10 ng·mL~(-1)) group, and low-(12.5 mg·L~(-1)), medium-(25 mg·L~(-1)), and high-dose(50 mg·L~(-1)) curcumol groups. The effect of curcumol on the viability of hepatic stellate cells induced by TGF-β1 was detected by the MTT assay kit. The apo-ptosis in each group was determined by flow cytometry. Real-time fluorescence-based quantitative PCR(RT-PCR) was employed for the detection of mRNA expression of α-smooth muscle actin(α-SMA), type Ⅰ collagen(collagen Ⅰ), and type Ⅲ collagen(collagen Ⅲ). Western blot was used to detect the protein expression of p62, microtubule-associated protein 1 light chain 3(LC3), beclin1, B cell lymphoma 2(Bcl-2), and Bcl-2-associated X protein(Bax). Transmission electron microscopy(TEM) was used to observe cell morphology and autophagosome formation in each group. The autophagic flux was observed after cell infection with adenovirus under double fluorescence labeling. The cell viability assay revealed that compared with the TGF-β1 group, the curcumol groups showed significantly decreased cell viability. The apoptosis assay showed that the apoptosis rates of the curcumol groups were significantly higher than that of the TGF-β1 group. RT-PCR indicated that the mRNA expression of α-SMA, collagenⅠ, and collagen Ⅲ in the curcumol groups was significantly lower than that of the TGF-β1 group. Western blot showed that the expression of p62, LC3, beclin1, Bcl-2, and Bax in the curcumol groups was significantly different from that in the TGF-β1 group. As demonstrated by TEM, compared with the TGF-β1 group, the curcumol groups showed significantly increased autophagosomes. The detection of autophagic flow by the adenovirus under double fluorescence labeling showed that autolysosomes in the curcumol groups were significantly increased compared with those in the TGF-β1 group. Curcumol can induce the autophagy and apoptosis of hepatic stellate cells, which may be one of its anti-liver fibrosis mechanisms.

Research progress on role of traditional Chinese medicine in hepatic fibrosis based on miRNA-mediated activation of NLRP3 inflammasome / 中国中药杂志

In recent years, liver fibrosis has become a hotspot in the field of liver diseases. MicroRNA(miRNA)-mediated Nod-like receptor pyrin domain containing 3(NLRP3) inflammasome activation is pivotal in the pathogenesis of liver fibrosis. The present study mainly discussed the role of miRNA-mediated NLRP3 inflammasome activation in the pathogenesis of liver fibrosis. Different miRNA molecules regulated liver fibrosis by mediating NLRP3 inflammasome activation, including miRNA-350-3 p(miR-350-3 p)/interleukin-6(IL-6)-mediated signal transducer and activator of transcription 3(STAT3)/c-myc signaling pathway, miR-148 a-induced autophagy and apoptosis of hepatic stellate cells via hedgehog signaling pathway, miR-155-mediated NLRP3 inflammasome by the negative feedback of the suppressor of cytokine signaling-1(SOCS-1), miR-181 a-mediated downstream NLRP3 inflammatory pathway activation through mitogen-activated protein kinase kinase(MEK)/extracellular signal-regulated kinase(ERK)/nuclear transcription factor κB(NF-κB) inflammatory pathway, miR-21-promoted expression of NF-κB and NLRP3 of RAW264.7 cells in mice by inhibiting tumor necrosis factor-α inducible protein 3(A20), and miR-20 b-promoted expression of IL-1β and IL-18 by activating NLRP3 signaling pathway. Additionally, the anti-liver fibrosis mechanism of different active components in Chinese medicines(such as Curcumae Rhizoma, Glycyrrhizae Radix et Rhizoma, Aurantii Fructus, Polygoni Cuspidati Rhizoma et Radix, Moutan Cortex, Paeoniae Radix Alba, Epimedii Folium, and Cinnamomi Cortex) was also explored based on the anti-liver fibrosis effect of miRNA-mediated NLRP3 inflammasome activation.

Effect of microRNA-125b on proliferation, apoptosis and collagen expression of hepatic stellate cells / 中国药理学通报

Aim To investigate the effect of miR-125b on HSC proliferation, apoptosis and collagen expression. Methods HSCs were transfected with miR- 125b mimics and inhibitors, and then molecular biology methods were used to detect the expression of Collagen I and HI in HSCs; MIT method was employed to detect HSC proliferation, and flow cytometry to detect HSC apoptotic rate. Results Molecular biological testing found that miR-125b mimics inhibited the expression of Collagen I and HI, and miR-125b inhibitors promoted the expression of Collagen I and HI mRNA; miR-125b mimics inhibited the proliferation of HSC, and miR-125b inhibitors promoted HSC proliferation; miR-125b mimics promoted HSC cell apoptosis, miR- 125b inhibitor inhibited HSC apoptosis, the above experimental results were statistically significant compared with its negative control group ( P < 0. 05 ). Conclusions miR-125b has anti-liver fibrosis effects, which may be related to the regulation of the phenotype of hepatic stellate cells.

Effect of curcumin against capillarization of hepatic sinusoids and its mechanism / 中国组织工程研究

BACKGROUND: Capillarization of hepatic sinusoids is an inevitable part in liver fibrosis and cirrhosis, and is a characteristic lesion inducing portal hypertension. However, curcumin effects on the capillarization of hepatic sinusoids and the underlying mechanism remain unclear. OBJECTIVE: To observe the effect of curcumin (a natural polyphenolic compound derived from the rhizome of Curcuma longa)on the microstructure and secretion of hepatic sinusoidal endothelial cells(HSECs),and to further explore its intervention on sinusoidal capillarization and pharmacological action mechanism of anti-liver fibrosis and target sites. METHODS: The rat HSECs were cultured and divided into seven groups: blank control group received no intervention and cells in the other groups were activated by leptin, followed by treatment with nothing (model group), high-, medium- and low-dose of curcumin, colchicine and salvia miltiorrhiza phenolic acid B, respectively, for 48 hours. RESULTS AND CONCLUSION: Under scanning and transmission electron microscopes, with the increasing activation of leptin, the number of fenestrae in HSECs was increased and the aperture was decreased. Curcumin could increase and enlarge narrowed or disappeared fenestrae caused by leptin, attenuated the thickness and scope of extracellular basement membrane, and reduced the degree of capillarization of hepatic sinusoids in a dose-dependent manner. Real-time PCR and ELISA results showed that after activation of leptin, mRNA and protein expression levels of endothelin-1 and vascular endothelial growth factor in HSECs were significantly increased compared with the blank control group (P < 0.05), while the expressions showed a significant decrease after treatment with curcumin in a dose-dependent manner (P < 0.05). There was also a gradient reduction in the protein expression of endothelin-1 and vascular endothelial growth factor in HSECs treated with curcumin. Moreover, all above mRNA and protein expression levels in the high-dose curcumin group were significantly lower than those in the colchicine and salvia miltiorrhiza phenolic acid B groups. In summary, curcumin can significantly alleviate the sinusoidal capillarization, and thus delay the development of liver fibrosis, probably by down-regulating the expression levels of endothelin-1 and vascular endothelial growth factor.

Efficacy of inactivated autologous porous bone flap and BAM bone-induced artificial bone for repairing skull defect in rats / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the effect of BAM bone grafting combined with inactivated autologous porous bone flap in repairing skull defect in rats.</p><p><b>METHODS</b>Seventy-two Wistar rats with skull defect were randomly divided into control group, inactivated autologous bone flap group (AB group), BAM bone-induced artificial bone material group (BAM group), and inactivated autologous bone flap with BAM bone-induced artificial bone group (BAM+AB group). The bone healing was evaluated with micro-CT and the new bone formation was assessed with histological staining at 1, 2, and 3 months after modeling.</p><p><b>RESULTS</b>Inactivated porous bone flap combined with BAM bone-induced artificial bone effectively induced vascular and fibrous tissue regeneration and osteogenesis in the cranial defects. With the inactivated porous bone flap as the scaffold, BAM bone-induced artificial bone obviously promoted the restoration of the skull appearance in the rats with cranial defects.</p><p><b>CONCLUSION</b>Inactivated autologous bone flap group and BAM bone-induced artificial bone material can promote skull healing and restoration of the original skull appearance, and can be used for reconstruction of the local anatomy of the skull surface.</p>

Clinical characteristics of pediatric hemorrhagic fever with renal syndrome / 中国当代儿科杂志

<p><b>OBJECTIVE</b>To study the clinical characteristics of pediatric hemorrhagic fever with renal syndrome (HFRS), and to improve its understanding so as to reduce the misdiagnosis.</p><p><b>METHODS</b>A retrospective analysis was performed on the clinical data of 26 children with HFRS between January 2009 and December 2012.</p><p><b>RESULTS</b>The age of disease onset was mainly distributed between 7 and 14 years (23 cases, 88%), and the male-to-female ratio was 1.89:l. The clinical manifestations of pediatric HFRS varied. The early symptoms resembled those of a cold, and in the course of HFRS, most patients developed digestive symptoms such as vomiting and abdominal pain. The laboratory examinations usually implicated platelet changes, and the imaging examinations revealed polyserous effusions. The prominent complication was myocardial injury.</p><p><b>CONCLUSIONS</b>Pediatric HFRS mainly occurs in school-age children, more commonly in males. HFRS does not have typical clinical manifestations or symptoms, so it should be distinguished from cold or appendicitis at the early stage. When applying the fluid replacement therapy, the cardiac function should be carefully monitored in case of heart failure.</p>

Study on the molecular-epidemiological characteristics of HIV-1 in Shenzhen, 1992- 2008 / 中华流行病学杂志

Objective To investigate the epidemiological characteristics of HW-1 subtype in Shenzhen from 1992 to 2008.Methods 489 HIV-1 positive plasma samples were collected from 1992 to 2008 in Shenzhen.HIV-1 env genes were amplified by nested-PCR from RNA.Phylogenetic analysis was performed on data regarding the nucleotide sequence.Results A total of 464 sequences were amplified and genotyped.Data from this study revealed that CRF01_AE was a predominant HIV-1 subtype in Shenzhen (64.4％,299/464),followed by subtypes CRF_BC( 17.5％,81/464),B'( 14.7％,68/464) and B (2.4％,11/464).Subtype C (0.4％,2/464),A1 (0.2％,1/464),CRF02_AG(0.2％,1/464) and CRF06_cpx (0.2％,1/464) were also prevalent in Shenzhen.CRF01_AE and CRF_BC were predominant among heterosexuals,homosexuals and injection drug users,while B'was predominant among blood donors.Results from phylogenetic tree analysis showed that some of the HIV-1 clusters had been defined in CRF01_AE strains at different time or groups with different transmission routes.Cross-infections were also seen.Conclusion CRF01_AE was the predominant HIV-1 subtype in Shenzhen while CRF_BC,B,B',C,A1,CRF02_AG and a small amount of CRF06_cpx or recombinant subtypes were prevalent in this city.Different subtypes showed great variation in the process of epidemics.

Progress in research on the biological reason of male homosexuality / 中华医学遗传学杂志

Male homosexuality is a complex phenomenon which is universal and with unknown causes. Researchers believe that both biological and environmental factors have played a role in its pathogenesis. Researches focusing on genetics, neurobiology, development and endocrinology have made certain progress. In this paper, we have reviewed the biological causes of male homosexuality, which may provide clues for further research in this field.

Epidemic trend and evolution of HIV-1 CRF01_AE strains isolated from 1992 through 2008 in Shenzhen district / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the prevalent status of CRF01_AE strains of recombinant HIV-1 in Shenzhen and their source of infection in order to predict the epidemic trend and evolution.</p><p><b>METHODS</b>A total of 489 samples of HIV-1 positive plasma were collected from 1992 to 2008 in Shenzhen. HIV-1 Env genes were amplified by nested-PCR from RNA. Subtype analysis were performed on the nucleotide sequence data. CRF01_AE sequences were analyzed by phylogenetic methods and characterized by calculating the genetic distance.</p><p><b>RESULTS</b>A total of 300 CRF01_AE strain sequences were amplified, accounting for 64.5% of all genotyped samples of all the 465 samples. The CRF01_AE strains of recombinant in 1992 - 1999, 2000 - 2005 and 2006 - 2008 accounted for 56.8% (21/37), 68.4% (78/114) and 64.0% (201/314) of genotyped samples, respectively. And in these three periods of times, 52.4% (11/21), 43.6% (34/78) and 45.8% (92/201) were heterosexually acquired adults; 4.8% (1/21), 0.0% (0/78) and 22.4% (45/201) were homosexually acquired adults; 19.0% (4/21), 51.3% (40/78), 30.8% (62/210) were intravenous drug users, respectively.Phylogenetic tree analysis revealed that HIV samples from different period of time showed distinct aggregation in time and transmission as well as cross infection. The gene divergence rate of CRF01_AE strains in the three different periods of time were (8.783 ± 4.717)%, (11.054 ± 7.141)%, and (13.218 ± 4.080)%, respectively.</p><p><b>CONCLUSION</b>CRF01_AE is the major epidemic strains in Shenzhen, which is transmitted through heterosexual contact, MSM and intravenous drug users. The gene variation increased gradually as time goes by.</p>

Spatial and temporal patterns of primary and secondary syphilis in Shenzhen, China / 中华流行病学杂志

Objective To learn the spatial and temporal patterns of primary syphilis and secondary syphilis in Shenzhen and to provide evidence for carrying out further research on syphilis.Methods Primary syphilis and secondary syphilis cases among residents in Shenzhen between 2005and 2009(n=11 303) were geocoded at street office level (n=55) based on residence at the time of diagnosis. Both spatial and space-time scan statistics were used to identify clusters of street office by using SaTScan software. Results In the purely spatial analyses, clusters were seen in the junction of the Baoan district and Nanshan district (Xinan, Xixiang, Nanshan and Nantou street office) and in the region near Hong Kong (Dongmen, Shekou, and Futian street office), as well as in the other streets where entertainment industry was relatively developed (Longhua, Huafu, Huangbei and Cuizu street office). The clusters had not changed much in the first four years, but nine clusters appeared in 2009.Annually, the most likely clusters were located in Longhua (2005, P≤0.001, RR=3.34), Bamboo (2006, P≤0.001, RR=9.59), Huafu (2007, 2008 years, P≤0.001, RR values were 4.18 and 4.75)and Cuizu (2009, P≤0.001, RR=8.02). In the space-time scan analysis, we found 16 significant clusters, which were similar to the pure spatial analyses. However, regional difference were also found, with the most likely cluster was the Guiyuan street office in 2006. Conclusion Spatial and space-time scan statistics seemed to be effective ways in describing the circular disease clusters. We have had a better understanding on spatial and temporal patterns of primary syphilis and secondary syphilis in Shenzhen through spatial and space-time scan statistics of syphilis surveillance data in the recent years. The changes of spatial and temporal patterns of primary syphilis and secondary syphilis were also described by SaTScan software, which also provided useful reference for the preventive strategies on sexually transmitted diseases as well as on HIV. Useful information was also provided for financial investment and cost-effective studies.

Molecular epidemiological survey on HIV-1 among the injecting drug users population in Shenzhen / 中华流行病学杂志

Objective To study the prevalent status of human immunodeficiency virus-1 (HIV-1) subtypes in IDU (injecting drug users) population in Shenzhen and to study their source of infection in order to predict the epidemic trend and evolution. Methods 166 HIV-1 positive plasma from the IDUs was collected from 1996 to 2008. HIV-1 env genes were amplified by nested-PCR from RNA. The C2-V3 regions (450 bp) of HIV-1 env were sequenced for analyses. Phylogenetic analyses were performed on the nucleotide sequence data. Results Among 166 samples, there were 6 HIV-1 strains including CRF01_AE, CRF08_BC, CRF07_BC 3 circulating recombinant forms (CRFs) and B',C, A1 3 subtypes. Data from the genotype analyses showed that 65.06% (108/166) were CRF01_AE, 19.88% (33/166) were CRF07 BC_6.02% (10/166) were CRF08_BC, 7.23%(9/166) were subtype B', 0.60% (1/166) were subtype C and 1.20% (2/166) were subtype A1. Phylogenetic tree analysis showed that some of HIV-1 clusters defined in CRF01_AE, CRF07_BC and subtype B' in different time groups. Significant increase of gene distance in CRF01_AE and CRF07_BC strains in the three different periods. Conclusion CRF01_AE and CRF07_BC were the major epidemic CRF strains among the IDU population in Shenzhen while the subtype B', C, A1 and CRF08_BC were also circulating in IDU population in this region. The variation of all different subtypes was increasing through these years.

Detection of T lymphocyte subsets in patient with glioma / 中华神经医学杂志

Objective To explore the relationship between T lymphocyte subsets and both glioma malignancy and its prognosis, and determine a clinical immunologic index for evaluating preoperative glioma malignancy and its prognosis. Methods The data of 117 inpatients with primary intracranial tumors, including glioma (n=85) and meningioma (n=32), were retrospectively analyzed. Fluorescence-activated cell sorting (FACS) analysis was performed to detect the preoperative contents of T lymphocyte subsets on 32 patients with meningioma and patients with glioma, including 45 high-grade glioma (WHO, grade Ⅲ-Ⅳ) and 40 low-grade glioma (WHO, grade Ⅰ -Ⅱ); and then the differences of their immunologic indexes were analyzed. Based on the detection result of T lymphocyte subsets, patients with glioma were divided into two groups: CD4~+CD8~+<1 and CD4~+CD8~+>1. Follow-up for 3-5 years was performed and the survival difference of these two groups was analyzed. Results Patients with high-grade glioma showed a decreased ratio of CD4~+CD8~+ and an increased value of CD8~+ with significant difference as compared with patients with low-grade glioma (P<0.05); patients with high-grade glioma showed a decreased ratio of CD4~+CD8~+, and an increased value of CD8~+ with statistical significance compared with patients with meningioma (P <0.05); patients with low-grade glioma showed a decreased ratio of CD4~+CD8~+ with statistical significance compared with the patients with meningioma (P<0.05). Patients with glioma showed a decreased ratio of CD4~+CD8~+ and CD4~+, and an increased CD8~+ with statistical significance compared with patients with meningioma (P<0.05). After follow-up for 3-5 years, 48 patients with glioma was found in the CD4~+CD8~+>1 group with 21 death (43.8%) and 31 months as a median survival time; 37 patients with glioma was found in the CD4~+CD8~+<1 group with 23 death (62.2%) and 16 months as a median survival time. The Kaplan-Meier survival curves were analyzed with statistical significance (P<0.05). Conclusion The prognosis is poor in patients with low ratio of CD4~+CD8~+. The preoperative level of T lymphocyte subsets in peripheral blood, correlated to the glioma malignancy, can be considered as an index to evaluate the glioma malignancy and the prognosis in patients with glioma.

Analysis of the tandem-repeat polymorphisms in DC-SIGNR alleles among drug users population with or without HIV/HCV infection / 中华预防医学杂志

<p><b>OBJECTIVE</b>To study the distribution of DC-SIGN/DC-SIGNR alleles among drug user (DUs) populations with or without HIV/HCV infection in Shenzhen, and to evaluate the role of these alleles in the construction of genetic resistance to HIV or HCV and screen out the anti-HIV/HCV gene in Shenzhen.</p><p><b>METHODS</b>All 500 DU blood samples were collected from Shenzhen Detoxification Center, including 313 from injected drug users (IDUs). All samples were screened for HIV and HCV antibody by means of ELISA. The genomic DNA were extracted and amplified by PCR. The neck domain repeat regions of DC-SIGN/DC-SIGNR were sequenced directly from the PCR products to confirm the amplification for some samples and all positive PCR products were analyzed by agarose gel electrophoresis.</p><p><b>RESULTS</b>Of 500 samples, 97 were found HIV positive, all of which were IDUs and HCV positive. The total positive rate of HCV among all HIV negative DU was 57.57% (232/403), and it was 63.89% (138/216) among IDUs; in comparing to the 50.26% (94/187) of DUs with other manners there showed significant difference (chi(2) = 7.61, P = 0.0058). Among HIV + DUs, there was a higher proportion of patient with the DC-SIGNR 5/6 and 5/8 (Fisher's exact, P = 0.043 and P = 0.034) with statistical significance; there was no statistically significant difference between HCV + and HCV-DUs and no significant difference between IDUs and other DUs for the DC-SIGNR polymorphism.</p><p><b>CONCLUSION</b>The results might indicate that DC-SIGN/DC-SIGNR polymorphism might not influence the susceptibility to HCV. Genotype 5/6 might probably have a relation with HIV infection, but still need further investigation for the low frequency.</p>

Oral administration of insulin inhibits islet beta cell apoptosis and prevents diabetes in NOD mice / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effect of oral administration of insulin on insulitis beta cell apoptosis and diabetes in non-obese diabetic (NOD) mice, and to explore the mechanism of immune tolerance induced by insulin.@*METHODS@#Eighty-six female NOD mice were randomly divided into an insulin group (n=43) and a phosphate buffered saline (PBS) group (n=43). From 4 weeks of age, the recombinant human insulin (Humulin R) 1 mg (70 microL) was administrated in the oral insulin group and 70 microL PBS in the control group respectively, twice per week before 12 weeks of age and then once weekly until 30 weeks. Insulitis and beta cell apoptosis of islets were observed at 12 weeks. IL-4 and IFN-gamma in the sera were measured by enzyme linked immunosorbent assay (ELISA). The expression levels of I-Abeta(g7), IL-4, IFN-gamma, IL-1beta, Fas and TGF-beta mRNA of islets, and IL-4, IFN-gamma, TGF-beta mRNA of Peyer's patch were measured by reverse transcription-polymerase chain reaction (RT-PCR) at 12 weeks.@*RESULTS@#The incidences in the insulin group were significantly lower than those in the PBS group (55.6% vs 85.7% at 30 weeks, 70.4% vs 96.4% at 52 weeks, P<0.05). The insulitis scores in the insulin group were lower than those in the PBS group, but there was no statistical significance. Fas expression on islets and apoptotic beta cell rates in the insulin group were lower than those in the PBS group (P<0.05). In the insulin group, serum IL-4 levels were higher, and IFN-gamma levels were lower than those in the PBS group (P<0.05). The levels of I-Abeta(g7), IFN-gamma, IL-1beta and Fas mRNA transcription in islets and IFN-gamma mRNA transcription in Peyer's patch were both lower in the insulin group, and IL-4, TGF-beta mRNA levels were higher than those in the PBS group (P<0.05).@*CONCLUSION@#The specific autoantigen insulin may induce the immune tolerance and prevent the diabetes in NOD mice, but it cannot block the progression of insulitis. Oral administration of insulin can induce the regulatory T cells, and make Th1 to Th2 cytokine shifts in the system and islets, thus preventing the Fas-mediated beta-cell apoptosis and diabetes.

Human immunodeficiency virus type 1 strains epidemic in Shenzhen / 中国医学科学院学报

<p><b>OBJECTIVE</b>To study the epidemic status of human immunodeficiency virus type 1 (HIV-1) subtypes in Shenzhen and to study their transmission source and routes.</p><p><b>METHODS</b>HIV-1 env and gag genes were amplified by nested PCR from uncultured peripheral blood mononuclear cells (PBMCs) obtained from 122 HIV-1 carriers confirmed in Shenzhen. The C2-V3 region (about 450 bp) of HIV-1 env and P17/ P24 region were sequenced.</p><p><b>RESULTS</b>Among 122 samples, 6 HIV-1 strains including 3 circulating recombinant forms (CRFs) of CRF01_AE, CRF08_BC, CRF07_BC and 3 subtypes of B', B, C were found in Shenzhen, and the percentages were 45.1% (55/122) for CRF01_AE, 31.1% (38/122) for CRF08_BC, 6.6% (8/122) for CRF07_BC, 14.8% (18/122) for B' subtype, 1.6% (2/122) for B subtype, and 0.8% (1/122) for C subtype. The intragroup genetic distances were (4.455 +/- 1.478)%, (2.997 +/- 1.345)%, (4.380 +/- 2.024)%, (5.186 +/- 2.487)%, and (4.869 +/- 2.638)%, respectively. In comparison with the sequence of respective international strains 01AE. TH. 90. CM240, 97CNGX-9F, CN. 97. C54A, B. US. 83. JRFL, and RLA2, the genetic distances were (5. 228 +/- 0.823)%, (3.634 +/- 1.073)%, (4.233 +/- 1.119)%, (4.950 +/- 2.564)%, and (5.795 +/- 2.198)%, respectively. The major subtypes found in injection drug users (IDUs) were CRF07_BC, CRF08_BC, and CRF01_AE strains. CRF01_AE and B' strains were epidemic mainly in sexual workers.</p><p><b>CONCLUSION</b>There are 3 HIV-1 subtypes (B', B, C) and 3 CRFs (CRF01_AE, CRF08_BC, CRF07_BC) epidemics in Shenzhen. The predominant subtypes varies among different transmission routes. While CRF01_AE is predominant among sexual workers, CRF08_BC and CRF01_AE are major subtypes among IDU population.</p>

Inhibition of islet beta cell apoptosis and prevention diabetes by subcutaneous administration of insulin in NOD mice / 中南大学学报(医学版)

OBJECTIVE@#To investigate the effects of subcutaneous administration of insulin on insulitis,beta cell apoptosis and diabetes in non-obese diabetic (NOD) mice, and to explore the mechanism of immune tolerance induced by insulin.@*METHODS@#Sixty female NOD mice were randomly divided into insulin group (n=32) and phosphate buffered saline (PBS) group (PBS group, n=28). Insulin was subcutaneously injected with humulin N (60 microL, 6U)+IFA (60 microL) at 4, 12, 20, and 28 weeks respectively, while the PBS group received PBS (60 microL) + IFA (60 microL). Insulitis and beta cell apoptosis of islets were observed at 12 weeks. IL-4 and IFN-gamma in the sera were measured by enzyme linked immunosorbent assay (ELISA). The expression levels of I-Abeta(g7), IL-4, IFN-gamma, IL-1beta, and Fas mRNA of islets were measured by reverse transcription-polymerase chain reaction (RT-PCR) at 12 weeks.@*RESULTS@#The incidences in the insulin group were significantly lower than those in the PBS group (21.4% vs 71.4% at 30 weeks, 28.6% vs 85.7% at 52 weeks, P<0.05). The insulitis scores in the insulin group were lower than those in the PBS group, but there was no statistical significance. Fas expression on islets and apoptotic beta cell rates in the insulin group were lower than those in the PBS group (P<0.05). In the insulin group, serum IL-4 levels were higher, but IFN-gamma levels were lower than those in the PBS group (P<0.05). The levels of I-Abeta g7, IFN-gamma, IL-1beta and Fas mRNA transcription in islets were lower in insulin group, but IL-4 mRNA levels were higher than those in the PBS group (P<0.05).@*CONCLUSION@#The specific autoantigen insulin may induce immune tolerance and prevent diabetes in NOD mice, but it can't block the progression of insulitis. Subcutaneous administration of insulin can induce the regulatory T cells, and make Th1 to Th2 cytokine shifts in system and islets, thus preventing the Fas-mediated beta-cell apoptosis and diabetes.