Objective To investigate the effect of stem cells transplantation on immune function,liver function and related indexes in the patients with end-stage liver disease(ESLD).Methods A total of 163 cases of ESLD in Nanyang Municipal Central Hospital of Affiliated Hospital of Zhengzhou University were selected and divided into 2 groups by the randomized single blind method.Eighty-one cases in the control group were given the conventional symptomatic treatment,while 82 cases in the observation group received bone marrow mesenchymal stem cell(BMSC) transplantation based on the control group.The changes of immune function,liver function,alpha fetoprotein(AFP),rate of prothrombin activity(PTA) and plasma total protein(TP) level before and after treatment were observed in the two groups.Results The immune function indexes CD3+,CD4+,CD8+,CD4+/CD8+,TP and PTA levels after treatment in the observation group were significantly higher than those before treatment and in the control group (P＜0.05),while the levels of AFP,alanine aminotransferase (ALT),aspartate aminotransferase (AST) and total bilirubin (TBIL)were lower than those before treatment in the same group and the control group(P＜0.05).There was no statistically significant difference in complication occurrence rate between the two groups(P＞0.05).Conclusion BMSC transplantation for treating ESLD can improve the immune function,improves the liver function and reduces the AFP level.

BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into hepatic stem cells in a specific environment, and participate in the repair and reconstruction of liver function.OBJECTIVE:To investigate the therapeutic effect of autologous bone marrow mesenchymal stem cell transplantation on decompensated hepatitis B.METHODS:Eighty-four patients with decompensated hepatitis B were randomly divided into two groups:normal group (n=42) with symptomatic treatment, and oral entecavir, Fuzheng Huayu Capsule; stem cell group (n=42) with the left and right hepatic artery transplantation of autologous bone marrow mesenchymal stem cells (1×106/kg) based on conventional treatments. Degree of liver fibrosis, liver function and score on Model for End-Stage Liver Disease (MELD) system, 1-year survival rate of patients were detected and analyzed with statistics before and 12, 24 weeks after treatment.RESULTS AND CONCLUSION:(1) Liver fibrosis after treatment in two groups:hyaluronic acid, laminin, type Ⅲ collagen and type IV collagen levels after treatment were lower than those before treatment in both two groups (P < 0.05), while these indexes in the stem cell group were lower than those in the normal group at 12 and 24 hours after treatment (P <0.05). (2) Liver function:after treatment, decreased alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels were found in both two groups (P < 0.05), and albumin, cholinesterase, prothrombin activity levels were higher than those before treatment (P < 0.05). The alanine aminotransferase, aspartate aminotransferase, and total bilirubin levels in the stem cell group were lower than those in the normal group at 12 and 24 weeks after treatment, while the cholinesterase level was higher in the stem cell group (P < 0.05). (3) MELD scores:MELD scores were both decreased in the two groups after treatment (P < 0.05), and lower in the stem cell group compared with the normal group at 24 weeks after treatment (P < 0.05). (4) The 1-year survival rate was higher in the stem cell group (69%) than the normal group (50%; P < 0.05). To conclude, the use of autologous bone marrow mesenchymal stem cell transplantation in the treatment of hepatitis B can significantly improve the patients' liver fibrosis and liver function, and improve the 1-year survival rate of patients.

Objective:To study the expression characteristics of Dickkopf1 (DKK1) in different time and space during tooth development of the postnatal mice, and to provide the theoretical basis for clarifying the mechanism of Wnt signaling pathway in regulating the tooth development.Methods:The postnatal Kunming mice at days 0.5, 6.5, 12.5, 18.5, 24.5, and 30.5 respectively after birth were selected and divided into various groups by time,three in each group.The mice in each group were sacrificed and the paraffin sections of mandibular bone including the first molar were prepared at the thickness of 5 μm, followed by HE staining and immunohistochemical staining in order to detect the expressions of DKK1 in tooth tissue and periodontal tissue.Results:At 0.5 d after birth, the mandibular first molar tooth germ was in the bell stage.At 6.5 d the enamel development of mandibular first molar was almost completed, and the epithelium root sheath extended to the root direction.At 12.5 d the dentin development of crown was completed, with the root formatted about 1/3. At 18.5 d the root had formatted about 2/3.At 24.5 d the root had reached the full length.At 30.5 d the apical foramen was narrow, and the root development was basically completed.There was no DKK1 expression at 0.5 d, but it expressed in the odontoblasts and predentin at 6.5 d. From days 12.5 to 30.5,the expressions of DKK1 were positive in periodontal ligament, alveolar bone, and cellular cementum as odontoblasts, which were gradually increased with the prolongation of time.However, no expression of DKK1 was detected in the pulp.Conclusion:DKK1 shows regular expressions at different tooth developmental stages after birth, suggesting its potential role in the growth of dentin and periodontal tissues.

BACKGROUND:There are many postmenopausal women taking hormone, which leads to much loss of bone mass, further inducing fragility fractures. The studies on the hormone exposure combined with ovariectomy-induced osteoporotic model are still immature, and the related molecular mechanism remains unclear. OBJECTIVE: To establish the rat osteoporotic model induced by ovariectomy combined with glucocorticoid exposure and to explore the underlying molecular mechanism. METHODS: Thirty 3-month-old female Sprague-Dawley rats were randomly divided into blank control, sham and model groups (n=10 per group). The rats in the blank control group received no intervention; rats in the sham group were clipped off a little of coeliac adipose tissue; the model rats received bilateral ovariectomy and 4-week administration of glucocorticoid. RESULTS AND CONCLUSION:At 4 weeks after modeling, compared with blank control and sham groups, the model group showed significantly lower bone mineral density of the femur, number of bone trabeculae and bone volume/total volume, and significantly wider bone trabecular spacing. Additionally, the model group revealed the damaged bone trabecular structure and thiner cortical bone. The expression level of Runx2 was downregulated whereas both collagen type 1α1 and peroxisome proliferators activated receptor γ mRNA were upregulated in the model group. These findings suggest that ovariectomized rats exposed to glucocorticoid rapidly develop femur osteoporosis, maybe by downregulating the expression of Runx2, as well as upregualting collagen type 1α1 and peroxisome proliferators activatedreceptor γ mRNA.

Objective To explore the effects of L-carnitine combined with citicoline on the therapeutic effect of neonatal hypoxic-ischemic encephalopathy(NHIE)and serum nitric oxide(NO)and endothelin-1(ET-1).Methods A total of 95 children with HIE admitted to the hospital from April 2020 to January 2023 were selected and were divided into observation group(47 cases)and control group(48 cases)by random number table method.The control group was given citicoline,and the observation group was given citicoline combined with L-carnitine.The therapeutic effect was evaluated on 14 days after treatment,and the recovery time of original reflex,muscle tone and consciousness was calculated.The levels of oxidative stress indexes[malondialdehyde(MDA),superoxide dismutase(SOD)]and vascular endothelial function indexes(NO,ET-1)were detected before treatment and 14 days after treatment.Neonatal neurobehavioral score(NBNA)was used to evaluate the neurological function of the children,and the safety of the treatment regimen was ob-served.Pearson correlation analysis was used to analyze the correlation between NBNA and vascular endothe-lial function indexes.Results The total effective rate in the observation group was 91.49％,which was higher than 72.92％in the control group(P＜0.05).The original reflex recovery time,muscle tone recovery time and consciousness recovery time of NHIE children in the observation group were shorter than those in the control group(P＜0.05).There was no significant difference in NBNA scores between the two groups before treatment(t=1.225,P=0.224).After treatment,the NBNA score in the observation group was higher than that in the control group(t=6.223,P＜0.001).After treatment,MDA level decreased and SOD level in-creased in two groups(P＜0.05).After treatment,the level of MDA in the observation group was lower than that in the control group,while the level of SOD was higher than that in the control group(P＜0.05).After treatment,the levels of NO and ET-1 were decreased in both groups(P＜0.05).The levels of NO and ET-1 in the observation group were lower than those in the control group after treatment(P＜0.05).The adverse drug reaction rates in observation group and control group were 17.02％and 8.33％,respectively,and there was no significant difference between two groups(P＞0.05).Pearson correlation analysis showed that serum NO,ET-1 and NBNA score in NHIE children were negatively correlated(r=-0.546,-0.608,P＜0.05).Conclusion L-carnitine combined with citicoline could improve the therapeutic effect of NHIE,shorten the re-covery time of clinical manifestations,and improve nerve function,oxidative stress and vascular endothelial function without increasing drugs and adverse reactions.In addition,vascular endothelial function indexes are negatively correlated with NBNA score,which could be used as auxiliary reference indexes for judging NHIE.

Heat shock protein A9 (HSPA9), a member of the heat shock protein family, is a putative receptor for Tembusu virus (TMUV). By using Western blot and co-immunoprecipitation assays, E protein domains I and II were identified as the functional domains that facilitate HSPA9 binding. Twenty-five overlapping peptides covering domain I and domain II sequences were synthesized and analyzed by using an HSPA9 binding assay. Two peptides showed the capability of binding to HSPA9. Dot blot assay of truncated peptides indicated that amino acid residues 19 to 22 and 245 to 252 of E protein constitute the minimal motifs required for TMUV binding to HSPA9. Importantly, peptides harboring those two minimal motifs could effectively inhibit TMUV infection. Our results provide insight into TMUV-receptor interaction, thereby creating opportunities for elucidating the mechanism of TMUV entry.
Blotting, Western ; Heat-Shock Proteins ; Hot Temperature ; Humans ; Immunoprecipitation ; Peptides ; Protein Structure, Tertiary

Objective:To evaluate a new type of draw-bar skin stretcher in repair of full-thickness skin defects.Methods:From May 2015 to January 2019, 52 patients with full-thickness skin defects were repaired with a new type of draw-bar skin stretcher at Daping Hospital, Army Medical University. They were 40 males and 12 females, aged from 4 to 61 years (average, 37.1 years). Their skin was stretched for primary wound closure. When primary wound closure failed, skin stretching was performed again to close the wound depending on the wound condition. When the Pinch test was negative after skin stretching, the wound was sutured directly. In cases of positive Pinch test, a skin graft or flap was used to repair the remaining wound. At 12 months after surgery, scar contracture and size of skin graft or flap were observed and wound healing after skin stretching was evaluated in comparison with the original wound.Results:After skin stretching, one-stage wound closure was achieved in 36 cases and multi-stage wound closure in 8 cases; of the remaining 8 cases, 2 were repaired by skin graft and 6 by skin flap after their wounds were reduced by skin stretching. In one-stage closed wounds, infection occurred in 3 cases and marginal necrosis in 5 cases; in the wounds repaired by skin graft or flap, no infection or necrosis was observed. The 12-month follow-up for all the patients showed fine healing of all the wounds after one-stage or multi-stage closure, linear scar, absence of scar contracture, and smaller wound sizes than the original ones after skin graft or flap repair.Conclusions:Skin stretching using our new type of draw-bar skin stretcher is an effective treatment for skin wounds. It can replace traditional skin grafting and flap surgery in some cases, but its indications should be strictly followed to avoid related complications.

Objective: To investigate the effect of papaverine on wound healing of full-thickness skin defect after skin stretching in swine. Methods: Eight Bama pigs were prepared. Standard full-thickness skin defect for 7 cm×7 cm was produced in the middle part of each pig's forelimb, and skin traction was used to close the wound. According to random number table method, the pigs were divided into experiment group and control group, with four pigs in each group. Percutaneous oxygen partial pressure (TcPO₂) was compared preoperatively and postoperatively. Wound healing clinical score was evaluated four and 10 days after operation. Weidner counting method was employed to determine the microvascular density (MVD) of the wound edge tissue 0 and 10 days after operation. Immunohistochemistry was used to observe the expression of hypoxia inducible factor 1-α (HIF- 1α) and vascular endothelial growth factor (VEGF) 0, 4 and 10 days after operation. Results: There was no statistically significant difference in TcPO₂ between the experiment group [(55.1±5.4)mmHg] and the control group [(54.7±5.9)mmHg] before skin stretching (P>0.05), while statistically significant difference was found after stretching [(22.7±3.3)mmHg vs. (16.4±3.4)mmHg] (P<0.01). There was no statistically significant difference in clinical wound healing scores between the control group and experiment group 4 days after skin stretching [(2.6±0.7)points vs. (2.1±0.4)points] (P>0.05), but better result was found in experiment group compared with the control group in 10 days [(1.3±0.5)points vs. (4.6±0.7)points] (P<0.01). MVD of skin tissue showed no statistically significant difference between the control group (8.5±1.3) and the experiment group (9.3±1.2) immediately after skin stretching (P>0.05), while the MVD of wound skin tissue in the experiment group (29.5±4.8) was significantly higher than that in the control group (21.1±3.4) 10 days after skin stretching (P<0.01). After skin stretching, the HIF-1 alpha expression in experiment group was significantly higher than the control group 4 days after skin stretching [(50.0±7.0) vs. (38.6±7.0)] (P<0.01), but no statistical significance was found between that at 0 day and 10 days(P>0.05). The expression of VEGF in the wound skin tissues of the experiment group was significantly higher than that of the control group [(49.9±10.8) vs. (39.2±6.3)] at 4 days after skin stretching (P<0.05), while no statistically significance was found at 0 and 10 days (P>0.05). Conclusion For full-thickness skin defect after skin stretching in pigs, papaverine can alleviate small vessel spasm, improve skin edge oxygen supply, and promote the growth of microvessels, thus promoting the healing, which might be associated with the increase of HIF-1ɑ and VEGF expression.

ObjectiveTo investigate the high-risk detection rate and aggregation of cardiovascular diseases（CVD） in 8 districts of Shanghai and influencing factors， and to provide scientific references for prevention and control of CVD. MethodsBased on the Cardiovascular Disease Screening and Management Program in Shanghai from 2016 to 2021， 104 685 participants aged 35 to 75 in 8 districts of Shanghai were selected for analysis. χ² test and multivariate logistic regression were used for statistical analysis of the influencing factors of CVD and aggregation of CVD. ResultsThe proportion of high-risk CVD individuals in the population was 19.17%， including the high-risk individuals with hypertension （8.65%）， dyslipidemia （6.33%）， CVD history （5.58%）， and WHO assessed risk ≥20% types （2.69%）， respectively. Old age， overweight and obesity， central obesity， smoking， drinking， farmers， unmarried， and low family income were the risk factors of CVD， while high education level was the protective factor. In the participants， 16 323 people （81.34%） were classified as CVD high-risk groups； The number of aggregation of 1， 2 and ≥3 high risk types of CVD were 16 323（81.34%）， 3 236（16.13%）， 509（2.54%）， respectively. Old age， low education level， low annual family income， farmers， unmarried， smoking， drinking， overweight， obesity and central obesity were associated with the risk of aggregation of high risk types of CVD， and the correlation strength increased with the increase of aggregation types. ConclusionThe prevention and control of CVD in Shanghai should focus on the hypertension， elderly， overweight， obesity， central obesity， smoking， drinking， low educated， low family income， farmers and unmarried people， and targeted intervention measures should be taken to reduce the risk of CVD among residents.

【Objective】 To investigate the relationship between serum reproductive hormones and sperm parameters and outcomes of micro-testicular sperm extraction (Micro-TESE). 【Methods】 Clinical data of 1 091 patients treated in our hospital during Jan. and Dec.2021 were retrospectively analyzed. According to the sperm concentration,the patients were divided into non-obstructive azoospermia (NOA) group (group A,n=418),normal sperm concentration group (group B,n=615),mild to moderate oligospermia group (group C,n=18),severe oligospermia group (group D,n=18),and obstructive azoospermia group (group E,n=22). In group A,244 cases treated with Micro-TESE were grouped into the sperm-acquired group (Micro-TESE positive group,n=82) and non-sperm-acquired group (Micro-TESE negative group,n=162),and according to the pathological types of testicular tissue,the patients were divided into normal testicular tissue with hypospermatogenesis group (HYPO group,n=129),maturation arrest group (MA group,n=10),and support-only cell syndrome group (SCO group,n=122). Differences in semen parameters and reproductive hormone levels were compared,and relationship between reproductive hormones and sperm parameters and Micro-TESE outcomes was determined with Pearson correlation analysis. 【Results】 In the sperm concentration subgroup,the testicular volume of group A was lower than that of group B and group E (P<0.05); the levels of follicle stimulating hormone (FSH) and luteinizing hormone (LH) in group A were the highest (P<0.05),but the level of testosterone (T) was the lowest (P<0.05); the levels of anti-mullerian hormone (AMH) and serum inhibin B (INHB) in group A were lower than those in group B and group E (P<0.05),the normal sperm morphology rate in group B was higher than that in group A and group E (P<0.05); the percentage of forward moving sperm in group B was the highest (P<0.05). Pearson correlation analysis revealed that sperm concentration,normal sperm morphology rate,and percentage of forward moving sperm were negatively correlated with age,FSH,LH (P<0.05),and positively correlated with testicular volume,T,AMH,and INHB (P<0.05). NOA patients were grouped according to testicular histology and pathology. The INHB in the SCO group was the smallest of the three groups (P<0.05); the FSH and LH levels in the SCO group were higher than those in the MA group (P<0.05),while the 17β-estradiol (E2) levels in the HYPO group were higher than those in the SCO group (P<0.05). NOA patients were grouped according to the results of Micro-TESE surgical treatment. There was a statistically significant difference in AMH and INHB levels between the Micro-TESE positive and negative groups (P<0.05). The binary logistic regression analysis of factors affecting the Micro-TESE outcomes of NOA patients showed AMH was negatively correlated with the Micro-TESE outcome (OR=0.904,95%CI:0.91-1.08,P<0.05). 【Conclusion】 Age,FSH,LH,AMH,and INHB are correlated with sperm concentration,normal sperm morphology rate,and percentage of forward moving sperm. The INHB level was the lowest in the SCO group. The results of Micro-TESE in patients with NOA can be predicted by serum AMH level.