To compare the therapeutic effects between acupuncture plus sacral injection and simple acupuncture for intervertebral disc hemia.Methods:The patients with lumbar intervertebral disc hernia at the age of 30-45 years old were divided into No.1-80 upon their visiting order,with odd number as Group A and even number as Group B.Group A was treated by acupuncture plus sacral injection,and Group B was treated by simple acupuncture.Results:The clinical effective rate was higher in Group A than in Group B,with significant difference (P＜0.05).Conclusion:Acupuncture plus sacral injection had the better therapeutic effect than simple acupuncture in treating lumbar intervertebral disc hernia.

Objective To investigate the significance of detection of Serum anti‐Helicobacter pylori(HP)and pepsinogen(PG) in patients with reflux esophagitis(RE) .Methods 118 RE patients (RE group) ,60 patients with other other gastropathy (other gas‐tropathy group) and 60 healthy subjects (healthy group) were detected serum HP antibodies ,PG levels ,compared serum PGⅠ ,PGⅡ level ,PGR and HP antibody positive rate between the RE group ,the other gastropathy group and the healthy group ,between the patients at different pathological degrees of the RE group .Results The levels of PG Ⅰ ,PGR and HP antibody positive rate of the RE group were lower than those of the other gastropathy group and the healthy group ,the healthy group was lower than the other gastropathy group ,the difference was statistically significant (P<0 .05) ,and compared with the control group ,there was no signifi‐cant difference in serum PG Ⅱ levels (P>0 .05);The serum PGⅠ level ,PGR and HP antibody positive rate of patients in grade A , B of the RE group were lower than those of the patients in grade C ,D (P<0 .05) ,the difference was statistically significant ,and there was no statistical significance in serum PG Ⅱ levels of the patients in grade C ,D (P>0 .05) .Conclusion The serum PGⅠlevel ,PGR and HP antibody positive rate of the patients with RE decrease ,HP infection is a protective mechanism of RE ,the dis‐ease exacerbates with the infection decreases ,detection of serum HP antibody and the level of PG has important clinical value in the diagnosis and prognosis of RE .

Objective To investgate the changes of bone mineral density (BMD) and biochemical markers of bone metabolism in elderly male patients with osteoporosis (OP) . Methods BMD and bone mineral content (BMC) of the lumbar spines (L 2～4 ) and the biochemical markers of bone metabolism in serum and urine were measured. Results BMD and BMC in OP group was reduced by 21.6% and 25% respectively vs that in control group. Bone formation indicators of serum levels of ALP and BGP in OP group were increased by 25.4% and 222% respectively vs that in control. At the same time, bone resorption indicators of urine Hop/Cr and NTX/Cr ratios were significantly increased by 22.6% and 223% in OP group than that in control. Serum T level of OP group was lower than that in control group and serum 25 OH D3 was below the normal value in both groups. Conclusions BMD and BMC of the lumbar spines (L 2 4 ) are the main basis for diagnosing the osteoporosis in men. Some elder male patients with osteoporosis are due to high turnover of bone metabolism. Androgen plays an important part in maintaining the bone mass. Vitamin D deficiency in elder men is the main reason occurring osteoporosis.

BACKGROUND:Fetal bovine serum based media used for expanding and cryopreserving human mesenchymal stem cells raise safety concerns in the clinical setting. OBJECTIVE:To investigate the feasibility of human umbilical cord blood plasma as a replacement for fetal bovine serum in culture and cryopreservation of human mesenchymal stem cells derived from umbilical cord. METHODS:Umbilical cord blood units were suitable for this research if they fulfil ed the donor selection criteria of the Guangzhou Cord Blood Bank strictly. Cord blood plasma was ready to use after col ected from the plasma reduction during the suitable cord blood units processing and pooling. Umbilical cord mesenchymal stem cells were harvested from the umbilical cord tissue of health ful-term newborns after delivery by enzyme digestion and were cultured in the presence of Dulbecco’s modified Eagle’s medium/Ham’s nutrient mixture F-12 containing either fetal bovine serum or pooled cord blood plasma. Morphology, proliferation, immunophenotype detected by flow cytometry and differentiation toward adipogenic and osteogenic lineages were utilized for investigating the effect of media on umbilical cord mesenchymal stem cells after 3-5 passages. Then cells were cryopreserved in media containing 10%dimethyl sulfoxide, 20%fetal bovine serum or 20%pooled cord blood plasma for at least 6 months. Viability, adhesion, proliferation, immunophenotype and osteogenic differentiation of the cells were assessed after thawing. RESULTS AND CONCLUSION:The morphology (spindle-shaped and plastic-adherent), phenotype and differentiation potential (osteogenic and adipogenic) were almost indistinguishable between cells cultured in fetal bovine serum or cord blood plasma medium, while cells grown in cord blood plasma medium demonstrated significantly higher proliferation rates than those in medium containing fetal bovine serum. After thawing, the cells maintained their adherence to the culture surface and differentiation potential to osteoblasts, but cells from cord blood plasma cryopreservation medium showed significantly better plastic attachment and produced greater cellnumbers than fetal bovine serum for the first three post-thaw passages. The results demonstrate that cord blood plasma can sever as an effective substitute to fetal bovine serum for growth, maintenance and differentiation of umbilical cord mesenchymal stem cells, and thus it wil be a safe choice for clinical-scale production of human mesenchymal stem cells.

Objective To verify whether the storage bag can reach the use requirement of cord blood freezing storage by conduc-ting a series of tests before staring use of new type cryopreservation bag for cord blood.Methods According to the standard opera-tion procedure,28 new type cryopreservation bags were extracted.The cord blood units were performed the cryostorage and thaw for rewarming according to the standard operating procedures.Then the physical integrity and various quality indicators of cord blood in bag were detected,including the nucleated cell viability,cell recovery rate,CD34-positive cell,colony-forming cultivation of stem cells,sterility test and verification of actual load volume.Results All the tested storage bags passed the integrity test,and the various testing indicators of the preserved cells conformed to the use requirements.Conclusion The new type storage bag has the same performance and effect to original bag,it is verified that the maximum loading volume of 50 mL(for 50 mL specification of storage bag)and 80mL(for 250mL specification of storage bag)is safe.

Objective To investigate the effect of phosphatidylinesitol-3 kinase/serine threonine kinase (PI3K/Akt) signaling pathway on expression of beta-site amyloid precursor protein cleaving enzyme-1 (BACE1) in the hippocampus neurons of rat brain. Methods Forty SD rats were randomly divided into 4 groups: blank control group, sham-operated group, insulin group and wortmannin group. Insulin or the specific inhibitor of PI3K, wortmannin was injected into hippocampus neurons to activate or inhibit the signaling pathway in insulin group or wortmannin group, respectively. Immunoprecipitation and Western blot were used to analyze the proteins levels of PI3K/Akt and BACE1. Results In insulin treatment group,among the proteins downstream of signaling pathway, expression of Akt increased (0. 952±0.060 vs 0.835±0.029,t=4.9150, P=0.0001), phospho-Akt set473 increased (0.800±0.075 vs 0.657± 0.025,t=4.5598, P=0.0002), phospho-GSK-3α decreased (0.604±0.062 vs 0.726±0.041, t= 3.5871, P=0.0018 ), and the expression of mature BACE1 and β-CTF significantly decreased. In wortmannin group, the expression of Akt and phospho-Akt ser473 were inhibited; phospho-GSK-3α increased ; mature BACEI (1.004±0.096) and β-CTF (1.031±0.048) increased (t=11.5980, P= 0.0000 and t =4.2194, P =0.0004, respectively). Conclusions PI3K/Akt signaling pathway might effect the expression of BACE1, in which impaired signaling pathway may cause the amyloid precursor protein to be easily processed by BACE1, and thus involves the pathology of Alzheimer' s disease.

Objective To investigate expression of β-site APP-cleaving enzymel(BACE1) and Aβ in brain of diabetes mellitus of Wistar rats,to study pathophysiological mechanism of Alzheimer disease from diabetic metabolic disorder. Methods Animal model of diabetes mellitus was established by streptozocin with intraperitoneal injection. Wistar rats were randomly divided into normal group (N), sham-operation group (S), 4 weeks diabetes mellitus model group (M4), 6 weeks diabetes mellitus model group (M6) and 8 weeks diabetes mellitus model group (M8). Behaviour was tested with Morris water maze and shuttle box test. Expression of Aβ was measured by enzyme linked immunosorbent assay and BACE1 by immunohistochemistry, enzyme linked immunosorbent assay, Western blotting and RT-PCR. The absorbance value was measured by imaging analysis. Results The electric times and latancy of memory and study were more increased in model group than that in N and S group but the times of escape more decreased(P<0.01). The expression of Aβ_(1-40) increased from (64.13±6.76)pg/mg in normal group to (86.43±7.03)pg/mg in model group by ELISA(P<0.001) and Aβ_(1-42) from (67.43±5.12 )pg/mg in normal group to (89.45±5.28) pg/mg (P<0.001) in model group. The expression of BACE1 increased from (116.46±8.10)pg/mg in normal group to (158.73±6.24)pg/mg in model group by ELISA and from 0.61±0.11 in normal group to 1.52±0.16 by Western blotting absorbance valule and from 1.62±0.26 in normal group to 3.61±0.32 by RT-PCR absorbance valule and from 0.81±0.21 in normal group to 2.01±0.36 by immunohistochemistry absorbance valule (P<0.001). The expression of BACE1 and Aβ in MT group was higher than that of in N and S group (P<0.01). The level of BACE1 and Aβ had positive correlation with cognitive impairment.Conclusion The expression of BACE1 and Aβ is increased in diabetes mellitus rats. Diabetes mellitus contributes to Alzheimer diseases that.

Abstract： Objective　To analyze the characteristics and corresponding drug resistance of pathogenic bacterial spectrum in eight major infection sites of hospitalized patients, and to provide epidemiological data for the rational selection of antibiotics in clinical practice. Methods　A total of 396 bacterial strains isolated from clinical specimens of hospitalized patients in member institutions of the Hainan Provincial Bacterial Resistance Monitoring Network from September 1, 2020, to September 30, 2022, were included in this study. Data were screened and filtered from the database of MH120 Microbial Identification and Drug Sensitivity Analysis System based on the technical scheme of the National Bacterial Drug Resistance Surveillance Network and Science and Technology Basic Resources Investigation Project research plan in 2020. The testing data were integrated, summarized, and analyzed using EXCEL and WHONET 5.6 software, and statistical analysis was conducted using SPSS 26.0 software. Results　Among of 396 strains of bacteria, 78 (19.7%) were isolated from respiratory tract specimens, 74 (18.7%) from urinary tract specimens, 72 (18.2%) from blood specimens, 54 (13.6%) from abdominal cavity specimens, 48 (12.1%) from skin and soft tissue specimens 48 strains (12.1%), 30 (7.6%) from reproductive tract specimens, 22 (5.6%) from central nervous system specimens, 18 (4.5%) from digestive tract specimens. Gram-negative bacteria accounted for 69.4% of the isolates, while gram-positive bacteria accounted for 30.6%. The top five gram-negative bacteria isolated were Klebsiella pneumoniae (14.9%), Escherichia coli (14.4%), Pseudomonas aeruginosa (10.4%), Acinetobacter baumannii (5.3%), and Salmonella species (4.5%). The top five gram-positive bacteria were Staphylococcus aureus (11.1%), Streptococcus agalactis (7.8%), Enterococcus faecalis (3.0%), Enterococcus faecium (2.8%), and Streptococcus suis (1.8%). Respiratory failure and bloodstream infection were independent influencing factors of treatment response (P<0.01). The resistance rate of Escherichia coli to ampicillin was 81.4%, and the resistance rate of Staphylococcus aureus to gentamicin and levofloxacin were both below 7%. Conclusions　The pathogen spectra vary with different infection sites of patients, and rational selection of antibiotics based on drug susceptibility testing is crucial to shorten the treatment time of patients and avoid the unnecessary emergence of drug-resistant strains caused by drug abuse.

Objective To design and synthesize novel 2-indolone derivatives as the c-Met kinase inhibitors. Methods With c-Met kinase inhibitor SU11274 as lead compound,a series of 2-indolone derivatives were designed according to the concept of bioiso-sterism. Then the target compounds(10a-10r)were synthesized from 2-indolone through 5-chlorosulfonation with chlorosulfonic acid, sulfonamidation with intermediate 3,condensation with 6a-6h,7a-7h and 4a-4b,respectively. Their inhibitory activity against c-Met and proliferation of MCF-7 cells were evaluated. Results and Conclusion The designed compounds were successfully prepared and their structures were confirmed by 1H NMR and ESI-MS. Some compounds had certain inhibitory activity against c-Met and prolif-eration of MCF-7 cells. An initial structure-activity relationship analysis of these compounds was performed to provide useful informa-tion for further optimization of their structures.

BACKGROUND:There are a lot of studies on isolation and culture methods of human placental chorionic-derived mesenchymal stem cells (hpcMSCs), but how to simply and efficiently harvest a large amount of primary MSCs has not been resolved. OBJECTIVE:To optimize the tissue explants method of isolating and culturing hpcMSCsin vitro. METHODS:Human placental chorionic villi were collected from full-term deliveries under aseptic condition and isolated by electric homogenizer. hpcMSCs were prepared by tissue explants method. The fluid and tissue of the primary culture flask and douching normal saline of the initial culture were centrifuged and prepared for secondary culture. RESULTS AND CONCLUSION: It saved time and effort to treat human placental chorionic villi with electric homogenizer, with good effects on tissue dispersion and removal of red blood cells. The average time of cell acquisition in initial culture and secondary culture was (17.73±1.14) and (10.03±1.30) days, respectively. The yields of primary cultured cells in initial culture and secondary culture were (6.97±0.98)×105 and (13.82±1.44)×105per Φ100 mm culture dish, respectively. The adherent cells showed fibroblast-cell-like shape, which were in parallel or circinate arrangement. Highly expressed CD73, CD105 and CD90 could be detected in the third generation of hpcMSCs, but CD34, CD45, CD14, CD19 and HLA-DR were negative. Following induction, alizarin red staining and oil red O staining produced a strong reaction in cells. In a word, the optimized method is a simple and efficient method for obtaining a large amount of primary hpcMSCs.